THE

JOURNAL

Copyright

OF HISTOCHEMISTRY

© 1975

by The

Vol.

AND CYTOCHEMI5TRY

Histochemical

Society,

POLARIZED

23, No.

1, pp.

21-50,

Printed

Inc.

LIGHT MICROSCOPY AS DIAGNOSTIC PATHOLOGY

A TOOL

1975

in LLS.A.

OF

A REVIEW MOSHE Departments

of Pathology

and

Cell Biology

and

Medical

Received Diagnostic as

Although been and

changes are being

lag

period

tic

procedures

mals is

and

will

their

introduction

diagnostic actually

which

made

polarized

help

in

light

line

of reasoning

help

and

Polarized visualize

objects

dark

field.

be

one

mainly

ferent words,

velocities at different the velocity of a ray runs

in

from

same

trajectory

plane. its

The laws

215)

and in

‘Established

Bureau,

nature

the

Ministry

simple

Investigator

of Health,

birefnin-

two

refractive

present

than

the

at

the

subject terms of

the

parallel that

to of the

to

than

that

Scientist

object

the

of the

polarizing

path

angle object

per-

Israel.

other

difference.

When tissue

will

which

of

structure

two and

fiber

An-

intensity

of the

light.

one

the

brighter

the

constitution plane of the

parallel

structure section are is twice

exhibit

double

as the

of birefningence. brightness also depends

is greatest of

polarization 21

the

retardation:

appears

thickness

the

same

between

a smaller

together, as

specific difference

determines

of the

the

of birefringence

is greater

is the identical in the

having

their the

with

which

of lie

sity

been

Chief

in the

intensity

indices

the

The polars

150,

for

with

fibers which

intensity

(252)

seen

objects

in the

along

thick

been

has

that index

perpendicular

are

in which

factor

examined

phenomenon,

(17,

runs than

is greater

objects

birefringence

the

have

realized

a positive birefringence. in a sphere means that

radius

accordance

other

dif-

through

used

rancause

refractive

which

a plane

Two

object

of an

of publications

recently rather

ray

is greater

Re-

causing usually

be

If the

polarized

differ

the

in a horizontal

of the

should

a sign.

in

of

birefningence.

of a fiber

RI of the

in

significantly runs

equipment

in a number

discussed

plane which

It

or

alignment

other factors molecules

of

fiber exhibits birefningence

a

azimuths. In other which is polarized

is polarized

physical

polarized

a

light

vertical of a ray

but

and

described 198,

the

on

in the

plane-polarized

that

ray

thickness

exhibit

characteristic

to

differs

shining

and

birefringence.

alignment

may

discussed

object

and

transmit

HINTS

of which,

is the

of the

polarity

pressure

microscope (“under crossed polars”) as shining bodies on a dark background. Different objects vary in their brightness, i.e., in the intensity of

can selectively or structures, and

mathemati-

parallel

consequently,

Birefningent

objects

the

tangent.

elements,

proper

Sheba

indicates

induce

has

length

the

tissues

Chaim

movement,

anisotropy.

axis, the Positive

include

PRACTICAL

bright

Birefningence

also

tissue

and

Anisotropic

will

review.

will

SOME

of phenomena,

gence,

of direct Following

with

of anisotropy

and,

the

diagnosis.

optics appear

in

or animal

microscopy compounds

good

number

review

AND

light anisotropic

these

was

familiar

often

of

(RI)

observa-

diagnosis.

human

help

BASIS

with

to

material

normal

might

THEORETICAL

and

on

recognize

thereby

human

the

made

which

addition

on

the

loss

15, 1974

Directional

birefningence

to human

microscopy

establishing

observations

in

not

laxation, heating dom arrangement

on experimental

May

form

presence

order.

molecules

review

to be applicable

are

stretching

ani-

School,

approach.

The

pathology

present

made

seem

pathology,

tions

this

the

observations which

on

to human

in revised

who

and

Medical

Israel

cal-physical

diagnos-

studies

University

and

sons

physicians.

of new

application

be

might

of

Tel-Aviv Hashomer,

16, 1974,

procedures have all the time, the

Accordingly,

include

animals

a group

and new introduced

Tel

January

group

in experimental

long.

often

as

a conservative

between

Center,

for publication

pathologists

regarded

WOLMAN’

Histology,

when

of

the image under crossed on other factors. The intenthe

object

45#{176} to the polars. parallel to one of (that

of the

is aligned

Alignment the two

polarizer

or

at an

of planes that

the of

of the

22

WOLMAN

analyzer

plates)

Lastly,

renders

most

birefringent refractive axes.

and indices

For of

nated

a

lying

in

the

NE-SW planes

South and East-West), as has in the preceding paragraph. appears refractive

dark when indices of

planes

which

pass

equal. Tilted ance with the and

the The

of

optical

through

axis

an

may

following:

molecules, form spatial arrangement

be due

of to

the

fractive

indices. on

interplay

of the

described

over

the between

compound crossed

a century

ago

sign,

effects

their

mounting tween the the

are

additive.

medium so that RI of the studied

medium

becomes

(3)

and

will in

and

a

of the

intensity

form

its

of the

birefringence

mounting object

and sum

two

can

medium

be

of similar between

birefringence many instances

is of practical weak and

birefringence

may the way

which

to

cal

succinic

peroxidase, niques which

(92) of the

may

linity

the

bireintense the of

obtained

by

a

RI

to that

using of the

be made

nature in which

importance as hardly observable easily

discernible

of the mounting medium. the weak birefringence

in by

spacing

here.

In

dye

which

moleincrease

chemical

are

moie-

aligned

a single

in a

plane.

reaction the

are

same

and benzidine

is

esterase and azo

a

hire-

true

for techdyes,

definite

spatial

enzymes,

or rather

crystal-

used

exhibit

dyes

polarization anisotropy

of

dichroism

colors by dyes

(43). and

The other

example, metals in impregnation has a similar histochemical mean-

of certain

reactive

(177, 189, arrangement

204),

and

groups.

importance of good, possibly strain-free and other requirements fur effective polight microscopy will not be discussed Mention

should

importance

of

embedded

be made,

proper

material. often

however,

dewaxing in

xylene

of’ the

of

Incomplete

occurs used

paraffin-

extract

sections (or

ion

treated other

some

structures

sections

than

others

of nuclei has

been

and

the

by

a

dewaxing is from

artifactual

in incompletely

described

of

by

fluid), or when the laboratory temperature low. Paraffin is extracted more rapidly birefringence

of

purposes.

ing to that of metachromasia which also depends on orderly

repeatedly

intrinsic

the

in

agents dichro-

products.

agents (for procedures)

The optics larized

Abolition

of the

indicate

commonly

and anomalous enhancement

case. In birefrin-

or

attached

phosphatase employ

of the

be seen

review, induce

diagnostic

that

arrangement

the

of this

within

and

respectively,

Many

to the

As will

dehydrogenase (55),

of

whether the observations that obtained in the histochemi-

was

represent

and

order

form

perpendicular

that

are

dichroism. absorption

molecules

they

definite

fringent

effects.

form

other indicates

paraffin

interplay

of

procedures

of the two planeto each other, the

alignment

It is not clear formazan deposits

studied.

The

changing Another

will

(or

certain

re-

increase

such intrinsic

orderly

appearance

different

other

for

the The

another

structure.

useful

anisotropy) ties

is

sections birefringence

often

addition,

the difference beobject and that of

signs, the compound the sign of the more

two,

two

the

even of

of

different colors) rays perpendicular

the subsequent which increase

the intrinof the same

gence have opposite fringence will have algebraic

observed

The

Changing

greater

intensity of birefringence those cases where form

and

of the

are

in the Such

of

and

a surface.

appearance

consists (or

cules

result

which

parallel

to and

of birefringence

as follows. Whenever form birefringence are

phenomenon,

axis

birefringence polars) of the

be summarized sic and the

This

the

one

propor-

the

in

are

to the plates)

varies

result

some

alignment

birefringence.

curves polarized

birefrin-

RI.

often

is

by histochemical

on

may

of

metals

orderly

products

the the

or both

is due (rods or

in the

deposition

increase

ism

is related groups

between

forms

half

axis

with enzyme

some

reaction

intensified

structures

of

result or

be markedly

certain

Impregnation

anisotropy,

in accordthe fiber

form

birefringence

two

as all

of a different

difference

The effect (that observed

out fiber

to one

and

birefringence of objects

form

tionally

or

examined

birefringence of atomic

in a medium

intensity

fiber

object

intrinsic

gence. While intrinsic the spatial arrangement

orderly

the

microscope.

of

polars

submerged

the

of the

in

can of

stages

reactions dyes

North-

in

staining

the

procedures.

(for

pointed same

cut transversely, polarized light

image

crossed

the

been The

structures

in

certain

polarizer

are

fibers appear brighter size at the angle between

bright

under

the

certain

illumi-

and

that

polarization

their spatial

fiber brightly

(assuming

analyzer

are

in

birefringent appears

directions

and

invisible. objects

exhibit differences along two of the three

collagen)

when

NW-SE

object

biologic

example,

example,

the

anisotropic

dewaxed Nedzel

(143).

POLARIZATION

Figure

1 shows

extracted

the

Weak

glass

that

of

the

of

sections 1.00

most

In

when

were

of

procedures

isotropy.

As and

the

has

effective

dure

of

(and

Allen

compensatory

Staining

by

an-

above, The

(2)

off has

and

some

to be par-

for

proceby

the

same

the

optimum

not

been

might

two

field

be

with in

used

not

FIG. 1. Amyloidotic spleen stained with Congo red photographed under crossed polars. Because of incomplete deparaffinization nuclei are intensely hirefringent. Amyloid in arteries is seen below center and near both upper corners. The importance of complete dewaxing of sections for good diagnostic work is obvious.

observing

birefringence

settings

pathology

were

increased

respect.

of

directions

nostic

this

weak

water

hire-

fuund

Nakajima

exposures

opposite

sign.

of was

intrinsic

explained

in

measuring)

photographic

and

were

and

in

1.53

studies

often

been

to dry

procedures

same

impregnations

ticularly

left

other

form

used

to

media)

These

routinely stains

in

of use

two diag-

in this

field.

isotropic.

The

sign

mined

of’ biref’ringence

with

however,

a

of

structures

son

in with

gent,

two

allows

studied

and

is

is

of

to

keep of

independent

exhibit detection

in

hirefringence under the

intrinsic they

(a)

both

by

deter-

posure, each hirefringent object and twice dark. The photograph gent

can

cancel

in

the

aligned

at

each

planes

dark

of

objects

the

to one

two

Thus,

under

indicate

polars. the

procedures

strating

after indicates

fibers

were

biref’ringence alignment. usual

independently The

trick

rotating

both that

one

used

the

polars object

by

of

procedure many

by

best

which

remained

the

both

some

de-

polarizer.

In

colors

may

Schnabel

(202)

of dichroism

and

in

glial

of tumors

A deflection

East-West

of

direction

pro-

results.

the

PATHOLOGIC

The

used

as

in

frozen

from

of change

is

rapid or

allows

an

active

and

the

last

fringent

way

in scar

the while

of

simple

collagen necrotic

collagen

can

tissue,

colla-

from

procedures,

sections,

a scarred

ELEMENTS

of’ differentiating

diagnostic

crvostat

croscopy

MESODERMAL

hirefringence

a quick

fur example

as

Even ob-

dark

AND

Collagen:

demon-

stems

45#{176}, lack

using

by

procedures. the

with effect

by

effect.

different

observed

subtractive)

tissue

circular

increased

in a study

normal

duced

be

anisot-

microscopists:

by

in

10#{176}from

polars of

fur

proposed

(or dichroic

about

gen,

Two

the

glia

after

to

a procedure

NORMAL

ap-

ropy.

jects’

such

(203).

obtained

analyzer

perpendicular

the

used

Schnabel

instances

the

additive

by

he

of

other

crossed absence

the

way

stained

must

or the

off

enhance

compound object

grees

use

An

in some rotating

Another

the

can

plate.

obtained

is twice lighted shows birefrin-

that

polarizing

and

this

signs,

the

invisible.

necessarily

for

opposite

be

polars

observa-

of

one

can

must

intensity

The

(b)

plane

objects of

not

object

as proposed dyes,

By

ex-

alignment. is

dichroic

mi-

polars.

photographic

their

including by

only

depends

and

nil.

45#{176} to the

renders

of

other

be

in a parallel

pearance

light

both

1:35#{176} during

procedure

Dichroism,

the

connecting

whatever

elegant

polarized

be of form, intrinthe form and the

are

may

Alignment

objects,

more

that

The

of sufficient given conditions

birefringence

birefringence

mind

birefringence

factors.

tion. This birefringence may sic or compound. Whenever

does

rotating

staining

detection

two

polar

work,

respec-

sign

a polarizing

a bar

birefrin-

the

built

with

Compari-

positively

identification

important

microscopic

be

blue,

directions.

(68)

croscope

structures.

It on

easy

Gustafon

deter-

birefringent

yellow which

the

an

Intensely

diagonal

collagen,

best

routine

allows

sign. appear

the

be

For

plate

the

may

tively,

can

compensator.

a gypsum

minati()n

23

PATHOLOGY

In

about

mounting

1.33.

fringence

easier

procedures. were

medium.”

mainly

made

versus

good

(RI

used

effective

been

following

(RI

“mounting

was

the

tissue air

and

DIAGNOSTIC

incompletely

of

has

any

studies so

IN

section.

anisotropy by

some air

appearance

paraffin

detect

MICROSCOPY

polarized different

tissue

on

light

iat

tuberculous network

necrosis. based

ion

between

focus, is

intensely

is

not.

mi-

as hire-

Polarized

in

24

WOLMAN

light

has

been

procedure

suggested

for

crossed

polars

collagen

Haq

Bagnell

and

splenic

arterial

is not

birefringent,

is often (31)

from

weak

hardly

other

phenol,

hyalins

played routinely

with

with central

It

a

is also

a

under

crossed

hydrated

those

of mature

mine

the

fibers can be easily distinguished from muscle (which shows birefringent

tion that fiber-water

transverse weakly

striation), birefringent)

Hatzel

weakly

or

not

smooth muscle (which is and elastic fibers (also

birefringent).

Collagen

is

;e’

--

-:

posi-

the

in the

less

(236) with

and

von with

of collagen which are of

diagnostic fibrils

is also to

which

aligned less

Puett, between

or not fihrils

bonds Tonna birefringence

age.

Engel

and (51)

infurma-

form b displacing (2:33), Tonna and

other

of

at all deter-

Elden

of available

are

than

Ciferri

of birefringence.

many

pathol-

of

perfectly

a review

fiber-fiber bonds.

intensity

creases

stained

cross-links

from

collagen striated

of sections

collagen,

intensity

(117.

by the examination

According

concluded

be

metallic

or silver

birefringence of factors

and

(166),

of the may by

copper

importance

Rajagh

a role in the total birefringence. stained or unstained sections,

birefringence

be obtained 198) or by

collagen,

more

of to a

(29).

intensity of on a number Young

length is due

birefringence

gold,

F:3BA

the

induced

may (194, polars

anisotropic.

that

of

dichroism

red

considerable ogy.

intense

possible

Specificity reaction

to

birefringence form

with

195). Ebner

The depends

in

more

and

and

picrosirius

a change

birefringence

into

birefringence.

of’ the

two acquired It is probable mainly to the

and

discernible

two

negative fibrin In

by

in phenol

relation

The

intensity

impregnations

hya-

hyalins of the

196).

intrinsic The

increased

hyalin

of birefringence the other treatment. are related

in

(23.

fibers. the

follicular

birefringent fibers

positive

Cooper,

follicular

of collagen fibers in the in the intense positivity hyalin

tively the

that

Mounting

intensity

useful Under

to contain 2).

shown

central

birefringent. the

follicular the

have

peripheral

central hyalin, while birefringence by this that all the findings presence decrease

shown (Fig.

while

a

change.

bundles and

tin is intensely decreased

hyalin

hyalin

numerous

as

(252)

studying

authors of and

noted collagen

Catchpole

that in(52)

-,

49_,

I

-

-A .,

I-. I

/-

(.

-

I

‘,

‘

/ /

Jo

I

W

I-

S

-

E

I,

‘U

.

FIG.

stained crossed

2. Old hyalinized tuberculous scar in pleura. section. Left, under ordinary illumination polars the tissue is seen to contain numerous

Photographs the hyalinized collagen

of the same area in a hematoxylin collagen appears amorphous;

fibers.

and right,

eosinunder

POLARIZATION

found

that

occurs

the

more

mals.

loss

course

and

in

years,

days.

that

the

most zero It appears,

occur

for

easily

discerned bands,

Laufer

et

studying

example as

of

gence

can

(101) polarized

run

of

resistance

From

parameters

the

author

3 days of

were

which

could

followed

by

aponeuare con-

of

imthe

birefrin-

of

guinea

of

pigs

training, and

over

nor-

increased reported the

to

these

that

de-

consequent

loss

caused molecular disorientation substance mucopolysaccharides be

gauged

decrease

by

in the

gen fibers. microscopy

Figure can

abnormal

alignment

loss

of

3 shows conclusively

how

of

of

collagen

size

above mentioned of collagen was

studying

pathologic

mahl

(125)

formalin-fixed formula gence

changes

standardized frozen

Retardation +

by

the the

phenol

disease

the

values

(phenol) useful in

in collagen.

Miss-

procedure and

change

developed in

birefrin-

of

dermal

by phenol

animals difwith Addicollagen

does

a

not

and

seem

Conti

the

to

(133),

keratocytes,

more

occurs

of

in the

of birefringent The second the

found

collagen

rounded

for example

(144) effect by

ground

that

cornea.

polardetect for

The

which

one

become

of AMP and occurs within

fibers,

where

structures

These structures are negatively birefringent phenol treatment.

in

birefrin-

them to characteristic

keratocytes

crystal-like

Neumark phenol

of the collagen some unknown unchanged

the

granules change

in

substance

intensely

Feh#{233}r(57)

dystrophy

between

the

For

although

became

Fran#{231}ois and

wounds

ground

light microscopy allowed two changes which are

macular full tein.

perforating

rabbits,

shape,

birefringent

treated collagen

in various In patients

ized the

of

the and

change

on

treatment:

of untreated collagen retardation of same fiber Retardation of untreated

The values obtained fered to a great extent. son’s

Ebner to be

the sections

representing caused

von found

biopsy

but the birefringence remained unchanged.

gent.

cardi-

omyopathy. The reaction

a skin

experimental

corneas

reason

light the

in

following swelled fibers

of colla-

polarized demonstrate

by

studying the birefringence of ground substance of cartilage stained with thiazine dyes, also found that these AMP are anisotropic. In the cornea of the eye, wound healing seems to proceed somewhat differently from the process in the skin. Varga and Feh#{233}r(238) found that the

anisotropy

orientation

disease

have been further tested. It has been noted that form birefringence depends on the RI of the mounting medium. In the case of collagen the nature of the mounting medium partly depends on the constitution of the interfibrillar matrix. De Campos Vidal (39) noted that the ground substance acid mucopolysaccharides (AMP) plays an important role in the form birefringence of collagen. The orderly alignment of the AMP of the ground substance of connective tissue could be also deduced from the intense birefringence of the AMP in sections stained by dyes mostly used as fluorochromes, such as acridine orange, trypoflavin and rivanol (187). M#{243}dis, M#{243}dis-Suveges

its

both

25

PATHOLOGY

fibers were appreciably higher than normal and the values returned to normal after corticosteroid treatment. The author proposed that the increased effect of phenol indicates that the collagen of Addison patients is immature and richer than normal in procollagen. In fact, the same change was found in the dermal collagen of children. This possibility of diagnosing Addi-

the

tendon

in comparison

(40) with

in

experiments

the

reduced

of tendons,

of tension, the ground

and they

the 7th day of exercise, Vidal

the

in healing

forced

progressively

tachment

shown

forces. The determining

the

After

iso-

began.

microscopy

intensity

from

is

or

DIAGNOSTIC

son’s

can be

healing

further

birefringence

De Campos

al-

of which

of collagen

This

to tear

normal.

RI

light

and

daily.

intensity

form was

fibers in tendons well aligned as

(173).

such (255)

birefringent

have

be deduced

Rollh#{228}user

wounds Gillman mostly

after

orientation

order

in

months

of wounds

exposed to mechanical of these forces in

degree

mal.

weakly

of

wounds. Collagen roses are always

stantly portance

during

(the

scars

months

al the

ani-

wounds of 1 month or less. that in sections mounted

even

usefulness

heating

in old

birefringence

RI of collagen),

tropic

IN

of collagen

slowly

in healing Wolman and changes involved

in healing therefore,

in glycerogel,

to

birefringence

Intrinsic

the

by than

while

take

birefringence.

near

in young

of aging

changes

found

of birefringence

rapidly

Changes

the

MICROSCOPY

proand

negatively can

be seen.

not collagen, as they and are not affected

observed that was reduced substance

in cartilage.

This

the intensity in collagen rich

are by of sur-

in AMP,

observation

might

as

26

WOLMAN

3. Irregular

FIG.

collagen

fibrosis is haphazard

network

have some pathology, changes Katenkamp

the

ground

(93)

rheumatoid

has,

of new

membrane. marked

This increase

decrease

in phenol the

birefringence

is associated and with

types

and

differ

collagen Scheuner

These

Rollhiiuser

studied

capillaries

and

of cerebral

form

birefringence.

Scheuner

reiter

(188)

the

ment

membranes

studied in

autopsy

the other

membranes blood vessels

basement small

known

to be of

the

Nies-

birefrin-

observed and

Hutschen-

birefringence

material.

They

collagen length.

effect

the

mahi

and and

positive

the cho-

negatively

was

This

inverted

indicated

that

which separate epielements contain perpendicular

of studying Hartwig

the

amnion,

to

Reticulin: Polarization microscopy used as a means of differentiating reticulin collagen

of on

of’ the

also

negativity

phenol.

aligned

reticulin (128)

birefringence

can

fibers.

observed

of

their be from

Missthat

reticulin

the

can

be

changed into negative birefringence by treating histologic sections with glycerin, or by mounting in glycerin-gelatin. The authors fuund that effect

is specific

be abolished Brewer (2:3)

by

fuund

that

quantitative

and sub-

be used to detect fibers. Treatment

chains reticulin

fur

arranged fibers.

with the

perpendicular Missmahl polarized

and

ret iculin

treatment

confirmed

base-

of capillaries in the intestinal

of

membranes mesenchvmal

fibers

negatively

length

no

were

their

with

of the directions.

membranes

tubules

of

for-

had

basement

hut

the basement thelial from

the

and

renal

treatment

were

to

epithelium and

organization in different

placenta

The

birefringent.

the

only

paraffin-embedded

malin-fixed

the

collagen.

collagen.

(145)

gence

zonula that

from

are

under

the

found

contain

the

and elastin (8:3) studof

and

of

the treatment

birefringence.

by

positive

refractile,

points

often

a

The oriented

respect

Phenol

non

with

the

in in

intestinal

collagen.

birefringence

and

in

a marked

that

and

birefringent

and

Therefore,

of cattle

mucosa

synovial

is lost.

both and

membranes:

Basal

the

seen

in essential

that

in the

with cardiomyopathy. thickened sheaths

vessel.

degeneration

easily

although

eyes

differed

different

on

of collagen

of the

fibers

sing

process of AMP effect

a patient and their

human with

is destruction

(basophilic)

intrinsic

ciliaris

found

fibers

polars from Hutschenreiter the

fact,

there

be

fibers,

crossed fibers. ied

in

of

constitution.

collagen

elastotic

it can

elastotic

myocardium muscle bundles

substance

arthritis

formation

In

the with

interesting applications in mainly in processes associated in

collagen

in

light

lipid

presence

that

it may solvents. of

lipid

to the length of (122) observed that microscopy

pathologic changes in with cortisone decreased

could these the

POLARIZATION

lipid

content

of these

negative

fibers

(the

in

sections

birefringence

glycerin)

in

the

ministration

liver,

of

increase

in the

in their

lipid

Elastin:

structure

of the

vitreous and 178) indicate

made

of

intrinsic

an

elastica

so

and nent

osteoid

The

FIG.

studies

4.

be

Intense

main

Figure

bireLngence

intensely

or

235)

..

the

the

total

with

the

orienta-

with

found by

Congo

and

Hartwig

mature

positive thors

indicate

gence

red.

could solution

,

erating

that

of that

be

both

and

the

he enand

Missmahl

hirefringence

intrinsic

and

last

the

sign

of the

by

treatment

.

Flor-

of human

The changed

c

and

Lugol’s

components. could

Divry

amyloid

found

to

intense positive he strengthened

Romh#{225}nyi (175)

found

due

mouse

consists

also

the

mostly

hirefringence

with

(126)

shown

the

staining

to

has

mineralization are

procedures.

that

experimental

of amyloid

(226)

deficient

that

contribute

Spreter

birefringence

staining

(45)

shown

organic

The moderately of amyloid may

with

birefrin-

the crystals

birefringence.

hanced

b..

increase

both

hydroxyapatite

compo-

J down

sections

Treatment

mucopolysaccharide-protein

hone:

in dentin

and

or in decalcified In decalcified hone

(234,

and

kin in

in tissue

blue.

to

Amyloid: birefringence

organic

formation to

et

network

in

collagen.

-

Modis of experimen-

prevailing

altered

a

either

that

acid

in

anisotropic

studied

4 shows

the

animals.

birefringence

shown

of

also in bone the intensely hire-

complexes. These complexes were again to play a role in callus formation. In dentin the situation is similar

by various

anisotropic

is also of Tonna

are

of

and that is more

toluidine was

is accompa-

in birefringence

of young

callus

with

fractures

increase

the

induced

changes

arterial

the

of bone

that

that

age. both

crystals can

the

is collagen.

gent.

bone

ground sections bone sections.

cartilage

noncalcified

with

In

than studied

by a

birefrin

human

fringent al. (132)

with

producing

of

to increase

birefringence

in

sheath The

layer

and dentin: and the apatite

undecalcified and mounted

outer

arrangement

outer

seems

that

separated

an

hirefringence.

the

studied

a longitudi-

proteins),

progressive

tion

with

in fibers

healing

by

vasopressin

Other elastic

component

the

nied

stained

27

PATHOLOGY

that

tally

has

while in appears

fibrillar. that the

(indicating from

intrinsic of

an

fiber

DIAGNOSTIC

newly formed collagen, collagen of old animals

change

in sheets, elastin

the

axial

of the

band micellar

negative

Bone matrix

an

sections

annular

gence

Ad-

light. Cruse (32) a hirefringent

hirefringence

in longitudinal isotropic

elastic

polarized elastin contains

and is (97, 176,

arrangement

without

by

different studies

nal

in

content.

is present nuchae

thin

of the

kidney. caused

extract

of fibers

component which the ligamentum

positive

IN

mounted and

a bacterial

The

is

intensity

spleen

amount

also been studied found that aortic

fiber

MICROSCOPY

form

mentioned

osteogenic

aubirefrin-

sarcoma.

with

WOLMAN

28 phenolic color to

compounds. of amyloid

indicate

an

molecules

on

The

green

stained

by Congo

orderly

alignment

the

amyloid

polarization red of

fibrils

(121),

cause of this phenomenon by Wolman and Bubis

has been (254). Figures

show

of

the

stained

The ent

appearance with

Congo

red

was

tion

dichroism

colors

various

have

dyes

One

(42,

of the

for diagnosing

and

been 123,

most

124,

the

obviously

crossed

red-stained

(95)

showed

positive

that

false

In view standard results,

positive

of the which

Wolman

need will

polars.

deposits

(253)

the

which

better

once definitely

the

Romh#{225}nyi (180, 181) ther tests for differentiating

with

was the study sections;

of the

are

and

for

ondary

not

to establish a eliminate false a pro-

cedure for staining of amyloid with toluidine blue and examination under polarized light. This procedure (STB) is less sensitive but more

mounting

The

media,

amyloid

stained while

negatively

birefringent.

the

two

and

that

after

the

Deposits

FIG. 5. Rectal biopsy of a patient suffering from amyloidosis. posits on the gland basement membranes. On the left diffuse, vessels. Congo red under crossed polars.

un-

demonstrated

by

of amyloi-

primary

author

found in

Congo

red

collagen

from

that

could

arabic, positively

similarly

difference be

sec-

in polar

gum is

stained

The

furcolla-

increased

is

between by

pre-

of the tissue sections with proteolytic The birefringence of amyloid stained red became more intensely positive

of collagen digestion.

secondary trypsin

amyloid

diagnosis established.

specifically

structures

treatment enzymes. by Congo

it

thin

recently published amyloid from

by

birefringent,

The

renders or

differentiating

amyloid.

by

procedures.

method

are

polariza-

procedures has been

although

demonstrated

of tiny

other dosis

gen

proposed

of

allows

which,

amyloid

detection

techniques

procedure

are

other

of differand the

procedures

results

amyloid,

and for

used

The

of substances

sensitivity

suitable

detection of the green polarization color was considered pathognomonic. It is common experience that the results of this procedure sometimes leave the observer in doubt, and Klatskin uncommon. restrictive

red

weak

currently

amyloid. not

Congo 6

other

elimination

tissues

observed used

deposits

in Congo

and

209).

commonly

amyloid

birefringence

easy

5 and

anomalous

repeatedly

than

diagnosing

dye

explained

orderly arrangement of molecules dyes attached to amyloid fibrils

consequent

for

the

amyloidotic under

selective

found

amyloid after of

more The

deposits

a mild

also were

permanganate

nonsecondary

On the right, amyloid deposits

intensely

author

mucosa situated

amyloid

with fine mainly

negative found

that

digested

by

oxidation. (primary

amyloid dein the blood

POLARIZATION

FIG.

and

Renal

6.

capsules,

generalized localized

amyloidosis arterioles and

myeloma, were not

trypsin green

after oxidation polarization color

Congo

red.

and

in the

endocrine though will be have

nervous

plaques

in

found the

the

is

and

core

staining

with (121)

amyloid

in

bodies

ously

reported

(136)

or

senile gives Congo

described the

fibrillar

tests,

microscopy the Alz-

and the senile plaques differ secondary amyloids. Cerebral are not usually demonstrated

polarized

Amyloid

Althey (45)

patients with presenile foci in the adrenal and to Katenkamp anl

(94), who used polarization a variety of histochemical

heimer fibrils primary and pora amylacea

of which

Hartwig

demonstrable

isotropic

light

to

structures,

amyloid

after

change of brains of dementia and in small other organs. According

the

their with

amylacea known

ocular

that

brain

Missmahl

similarly

Stiller after

or by

preserved staining

Corpora deposits are

system,

birefringence

red.

tumor digested

tissues, prostate and the lung. they do not belong in this section, described here. Divrv and Florkin already

green

and after

amyloids: amyloid

localized

occur

IN

DIAGNOSTIC

29

PATHOLOGY

in a patient suffering from familial Mediterranean fever. Glomerular arteries are severely involved. Congo red under crossed polars.

amyloidosis, senile deposits)

Ectodermal

MICROSCOPY

amyloid

of the

prostate

in

the

literature

(208).

As

from corby

procedures have as these

(88). been

vari-

anisotropic

bodies

repre-

sent

secretion

variable

inspissated

degrees,

the

and

capillaries

compressed

intensity

of their

to

birefrin-

gence varies; this explains the negative findings of some authors. The present author found that well

formed

prostatic

the

reactions

which

Fig. 7); the same amyloid bodies veterinary

pathology

mammary

Smooth

and

striated

for

which 8).

markedly muscle,

to

increases while the

striated

Conti,

years

is weaker

According

muscle

exhibit parently

muscle

Atrophy muscle birefringence. fringence

and

in

muscle

than

has

This

both the contribute

Fischer

is

a

intrinsic to the

that

of collagen

(54),

stretching

The

Gotzos cells

loss

the

of

wall

functional

(54)

the

of

indicates of

birefringence

affects

Fischer degeneration

observation

(30)

in

intense cells.

markedly

positive

The

19:3).

is negligible.

a more relaxed

of

the birefringence of smooth effect on birefringence of

M#{244}disand

stretched

common and in

amylacea

striated

birefringence and form birefringence

positivity (Fig.

and

many

all (see

(167).

muscle:

been

compound and the

corpora

gland

of smooth

exhibit

fur amyloid of the less alveoli,

of the

birefringence known

bodies

typical

is also true of pulmonary

the

bovine

amyloid are

that arteries

than

ap-

activity intensity

found atrophy

in of

its

reduced hireof striated

30

WOLMAN

FIG. 7. Amyloid bodies within dilated lumina of prostatic glands. The amyloid bodies vary in the intensity of birefringence and in their structural pattern. The bodies on the right show Maltese crosses and appear laminated. The amyloid body on the left of center and the numerous small bodies in the upper left appear granular or of radial pattern. The birefringent fibers in the septa are collagen.

cytoplasm has never hand,

has been gained

Orban

and

repeatedly acceptance.

Romh#{225}nyi (149)

fully used polarized light ing myoblasts in Wlms’ Myocardial

cells

exhibit

of

dine

blue-stained ischemic

Soviet fringence

FIG. 8. vvi 11 of human ileum. in ‘ thick intensely birefringent bundles on the right are collagen. The weakly birefringent bundle of loose fibers running through the center of the figure is smooth muscle.

muscle. omyomata broblastic weak

The and

possibility of leiomyosarcomata

counterparts birefringence

by of

the

differentiating from demonstrating smooth

muscle

the

intravenous 15 by

leifi-

heart

in

the

fibrin

fibers.

The

polarization

fuund cases

changes

in the

by

hirecon-

of

hire-

of given

epinephrine.

microscopy

of’ rheumatic caused

loss contraction of rats

or

could injection.

auricular

to detect

myocardium.

changes in in pathologic

cells of

birefringence mm of the

be used the

observed lysis

injections in

brinoid

cell

can in

237

caused by in myocardial

changes within found

sections changes

Tsellarius

fringence myofibrils

path-

fibers. Orban and such study of tolui-

pathologists described of myocardial cells

ditions.

scientists

for studying

ologic changes in myocardial Romh#{225}nyi (149) found that early

some phe-

the presperpendicu-

by

microscopy

bire-

that This

caused

polarization

to be

success-

positive

(120) fuund birefringence.

found

but it other

fur detect-

ence in the fibers of lipids oriented lar to the fiber length. A number used

was

have

microscopy tumor.

mostly

fringence, but Missmahl fibers show negative nomenon

suggested, On the

The

be observed Revzis (168) that

appendage carditis ischemia

the

fi-

of the contains in birefrin-

POLARIZATION

gence

of myocardial

possibly on rats and

cells

of two

types.

treated

isoproterenol,

(159)

found

IN

that

the

development

ated

with

changes

Kiefer

found

in addition

to

that

enamel

31

PATHOLOGY

that

or

Pilny,

DIAGNOSTIC

study

to be biphasic,

In an experimental

with

Sandritter

seem

MICROSCOPY

the

of dental

caries

in birefringence. intrinsic

is lower

birefringence

than

that

In an extensive study Gustafson (69) showed that polarized light

which

hereditary

birefringence

was

was

increased

normal,

and

(b)

eosinophilic

striation

but

with

increased

Fibrin: of fibrinoid from fungal respect

fibers

micelia. to the

both form pronounced (244)

in

and

contains the

a

most

brils

from

Angmar, however,

observed

hair: other

of

birefringent

The

An

species

are

the

extensive

of human by Schmidt

hair

was (192),

in diagnostic in

identification allows the

of keratin establishment

cyst,

are aspiration amniotic fluid found

which

in

an

of the

in

which

the

lungs

ectopic

cement;

of dental

when

length

of

and

cement

heated

Hoppe In

to be modern

Hoppe (81) birefringent

mature

the

the

apatite

ences

in packing

used

for

determining

could

not

penetrate

studies

pilonidal

is found

(6),

polarization mirole in the study caries. Study of

enamel

is over

noted than

it is negative prisms.

a number

are

been

also

to

found

of’ authors

not

be

aligned

or recently

obtained

by studies

correction alignment

of of

mineral

These

obser-

by

differmedia

birefringence

denser

areas. that matrix

that

In further the

intrinsic

of enamel the

in an ordered

of the variability in various parts

of intrinsic

immersion

found

indicating

associ-

the

caused

some

organic

birefrin-

different

in form birefringence has also been noted that the differences

in unerupted

in respect cement

low,

differences ous areas who found mation of order

the

of the

extremely

that enamel dentin and

Developing

to 800#{176}Cwere

100

that

intrinsic

author

the

form

areas.

to

the

dentino-

caries are birefringence

of the

so that

the

birefringence

in the

(with

birefringent,

in different believed

enamel study of

at the

by

found

and

were

birefringent

tissue

incipient in the

crystals varied

there

deminerCarlstr#{246}m that nega-

birefringence of enamel, even after the data for effects of the imperfect

vations

fluid in in moth-

zone

The intento vary in

layer

caused

(4)

or of hair diagnosis.

is

protein

mi-

fashion.

The

between variby Houwink (82) are also present

erupted

teeth.

Confir-

in density and degree of single teeth has also on acoustic

anisotropy

(104). by

positive. times

Angmar

lesion

of increased

(5) has shown, did not deter-

is positively mainly

Fluorosis and with changes

celles

birefringence

carious

this

the

of a thin

content,

and

hair

being

enamel.

the

lesion.

years old. In 1862 is more intensely

the

of mature crystals. The

enamel

junction)

effect

shown birefrin-

of birefringence. was found

the

of amniotic embolization

Enamel: In dental tissues, croscopy played an important of a number of diseases, mainly the

which

birefringence by the apatite

exception

gence. ated

medulla of the present, detecand keratin is

squames of a correct

instances in

center

for

of

published who later

conditions

are

that

of

of the

previously

Inside

the intensity of birefringence

enamel

tonofistudy

importance

practical except

squames

one

Carlstr#{246}m and Glas that mineral content

possible

structures,

of which

pathology

ers-both

In cases

by a zone

birefringence.

tissue.

positive

found

different areas. In developing

epidermis

animal

not

cases and

authors

intrinsic caused

mine sity

ELEMENTS

in the (200). At of hair

Such infants

had

exhibited

surrounded

negative

and

described lipid deposits hair of many humans tion of the birefringence of great

diseases. imperfecta,

co-workers

enamel

The

tive was

ECTODERMAL

(196).

in of

fibrin.

many

number keratin

the

and is more Von Dungern.

phenomena

and

of

the birefringence many years ago

instances

blue-stained

conspicuous

and

in some

components fibers.

keratin

humans

with

that

of dental

amelogenesis

authors gence.

birefringence.

PATHOLOGIC

Epidermis, of

of

birefringence is positive of the fibrils, consists

these

ANt)

over loss

in a number

are two areas of partial and complete alization associated with isotropy. (25) reviewed the subject and noted

in trypan

NORMAL

507i with

help

and

intrinsic stretched

described

dichroism

about cells

the

is sometimes

The length

and

by

and

normal

and Gustafson microscopy can

birefringence of fibrin and helps to differentiate them

The weak deposits

collagen

preserved

(35)

of carious

of the

frankly necrotic cells in which no striation or birefringence could be observed, there were two other types of changes: (a) eosinophilic cells in cross-striation

is associ-

Darling

noted

The observation fringence in the associated

with

of differences enamel of differences

a

in form hiresingle tooth,

in permeability

to

32

WOLMAN

immersion importance. and

media, Those

more

pected

may areas

be of which

permeable

to be also

than more

by various noxious as loci of lowered

great practical are less packed

others

can

accessible

agents resistance

ex-

and

Vidal

changes

and

Sudan ids are

tions

dystrophy

(240)

cells

black, indicating that arranged perpendicular

found

of the

observed

epithehial

Secreting Olivecrona

hal

cells

and

absorbing found

appear

anisotropic

isotropic

when

birefringence

was

direction

in

mounted

of the

negative lumen

in to

whole

cytoplasm

glands

secretory

while

secreting

crine

pancreas

the

was

frogs, sive

basal

some

weak

part

cells

In were

In the

birefringence

was

cells.

exhibited

In the

exoin

kidneys

in the brush the thyroid, and cumu-

lus oophorus

both

and

in

glycerin.

birefringence

The which

glycerin-mounted oriented perpendicular it could solvents. in

anisotropy authors could

renal

electron

that

detected

structures

and

microscopic

to

in

lipids axis, as

with lipid the findings

correlated

observations

the

only

sections was due to the apex-base

be eliminated by treatment Rollh#{228}user (174) extended

Sinapius, ported on

in water

found

be

them and

tals

in

The

with

triglycerides

function.

birefringence authors

layer

in which

the

terminal web. The was caused by the protein

filaments

regrettable been properly

that

In steroid-secreting

caused

found

a

anisotropy

by

deeper was

the

caused

birefringence of presence of axially with

these utilized

micro-

birefringent by the

this layer oriented

perpendicular

lipids.

observations in diagnostic

have not pathology.

organs,

the

appearance

their

needle-like

It is

of

nized seem

not

of birefrinis related to to

indicate (216) refat crysobtained

crystals

(3-40

heads of abnormal to have

in

the

published

other

been

appolar

on birefringence

disorganized

semen

of

crystals and

The possibilities of distinguishing from

type

form (see pages processes

These acids

fat of to be

second

spherocryst.als from hipolytic

autolysis.

been

seemed The

were

it),

within to consist

appearance

of fatty

yet

tumors in some

tissue

fixation.

during

sperm study

indireserve

precursors.

long

their

of spermatozoa (15). in these observations the

presence cells

in bundles, situated crystals were shown

crystals

have

presence

is believed

to consist

Studies

the functional crystals repre-

hepatic

granules or and resulted

peared lipids.

the

adrenal

and surgical biopsies. Two types were observed in hepatocytes.

anisotropic

needles, 33-35)

the

and Gunkel of anisotropic

by formahin

occurring

the

and

and

enhanced

tion

the

are

that

in

41), in ovarian villi (250) and

formalin-fixed

first

The

a functional

Avenarius presence

often arranged droplets. These

of

to

the

lipids.

of

of steroid

from autopsies of lipid crystals

Jahn, Scheuner and Hutschenreiter (87) studied the birefringence of the apical part of the small intestine epithelial cells of rats. In addivilli,

activity

accumulation

of

and later of Duguid and

these

testes (135) the in the secreting

functional the

(26,

and

adrenals

indicated

and

Mulon

in which excessevere infections

lipids

indication In fact,

in the

(157)

the

humans and

others

Also, in the ovary (61, 116), in placental of the lipids

(75).

on

precursors

also and

Hueck

birefringent

and

their

on

of Bennett (16) (38), Symington,

is a good of the cells.

cells gent

birefrin-

gent striae in their basal parts and borders. The epithehial cells of choroid plexus, liver, gastric mucosa exhibited

and

birefringent

mostly

ovary

Herrmann

reported

(231) of

lose

observations

by

of

and obserinfec-

were

the

cates that the cells contain and are not exhausted.

found

negatively

studies and Greep

cortex state

of

dogs and exercise

depletion

sent

salivary

anisotropic

negative.

of secreting

tubular

In

(248)

classical Deane presence

lipids lipids

Wacker

caused

Davidson

prostate, tube the

made

rabbits, muscular

the

membrane.

cells were

are

adrenal

Similar

were

had

1895,

extended this that in severe

lutea

Pickard,

The to

intestine, and

birefringent.

duct cells

relation

basal

the epithelial cells of the seminal vesicle, endometrium

but

glycerin.

corpora

Wehtmann

Hillarp epithe-

water, in

in

(137),

lipgingival

epithelia: that various

(77)

the

of of

cells in

Birefringent

adrenals

with

sudanophilic to the

poisoning

hypernephromata.

dichroic

stained

and

birefringence.

surface. and

adrenocortical Kaiserhing

occurred.

macular

Bozzo

in gingival

in by

Kaiserhing and Orgler (91) vation. The authors noted

surface epithelia: Fine anisotropic were found (57) in the corneal epithewith

droplets

discovered

may be regarded or areas in which

damage

cornea.

already

lipid

been

Other granules

of patients

the

to penetration

primary

hum

has

be

birefringent crystals or liquid crystals is part normal functional activity. The birefringence

and exploited.

inherent orgaones

male

sterility

in

POLARIZATION NORMAL

AND

PATHOLOGIC

MICROSCOPY

NERVOUS

IN

TISSUE

ress

Some

processes

involving

in the

nervous

and

39.

page

40.

Nerve

years and

system, syrup

disease

fibers:

It has

been

(96)

that

gent

the

system

in respect

to the

is

pages

length

on

for over

of the

fiber.

birefringence axial alignment

in

degeneration

nerves exhibit weak 201) which is caused

positive by the

the protein constituents. intense birefringence

In frozen sections the of myelin together with its

(66, of

has

under

respect identifying

to

circular

polarized

crystals polars wrong.

and

damaged

provide fibers. light

a rapid Schnabel

for

nervous

the

study

system

the

of myelin

sections polarized

in linear polarized light light. The identification microscopy myelinated

of

artifactual

tissues

Figure

in transverse and

in circular of myelin by

is important nerve emboli

squeezing

rounding

of normal

structures.

9 shows

polarization stances

appearance

means of (203) used

of nerve

in and

myelin

into

sur-

(27). is often reversible, light microscopy:

sure

cause

tension

may

material

may

consist

erides.

play

Positive polarized

ble

the

by

known

in zoology,

fragmentation nerve was cooled

Demyelination, process

whether

or as part

is associated globules and loss found

of the

myehin

as

of nervous

negative

occur

in an

early

the

when

using

a

autonomous destruction,

these 112). neuron

(190)

proposed

ordinary

and result, with time, Polarized light studies

in reversal of the prog-

cells: cells

strating

and

esteri-

role

in their

as proposed

The which

few

be

granules,

cell, and

which, method

SO3H

well

studied

(186, see also but specialized by polariza-

Scheuner

is based of

of easily

is have

neurosecretory

procedure

microscopic

moieties

existence can

workers

granules. a

light these

sulfur

oxidation stained

of 142,

depends hydro-

microscopy

but

is the

(225)

the

made (141, and

reaction

glial

microscopy

Spiegel

of

as Maltese

identification of free cholight microscopy is possi-

polarization

its specific

Wallerian

solvents or triglyc-

be

important

digitonin

and

rather

of

acids

appear

cells in human material Another type of normal which can be demonstrated

tion

birefringence

an

ganglion

into with

stage

It

(110).

of myehin eventually

birefringence.

changes at

an

tissue

with the breakdown myehin figures and

that

degeneration of the sign.

of myelin in vivo.

by

Neurons

Baldi

in lipid

fatty

light may lipid substances

constitution. lesterol with

myelinated

nerves and

for

in the tissue that sphero-

as triglycerides

often

detected

in teased

as a means

soluble which

constituents, sterols

reduced

narcosis,

be used

171, 197). Formation of spherocrystals on the presence of both hydrophilic

observed

during

are

in polarized of different

by Lison

birefringence

myehin

the

products

crystals acid and

Spherocrystals

crosses mixtures

(66). It has been found, however, this effect is obtained in axons and cells but not in myelin. Spiegel (224) in vivo

of

the

that

of the compounds. the tacit assumptions

acicular fatty

which

and

(10) found peripheral

birefringence

in

can

peri-

pulp.

breakdown

of cholesterol,

constituents (242) that Schwann

in nerves

of

changes

(20) in

giving Maltese crosses under crossed are cholesterol esters are completely Acicular birefringent crystals of endoge-

nous

fied

may pres-

sclerosis changes in

suggested

that

or

intensity

and

sign

birefringent cholesterol

phobic

Minor damage, which be detected by polarized or

inof

polars

birefrinin suda-

Almeida

dental

myehin

identification be stressed

normal

of myelin with con-

birefringence

repeatedly

crossed

that are

Bo#{128}o and of

of inflamed of

positive should

to

in multiple ischemic

in

153).

been

appearance

straight course (different from the wavy course of collagen), tubular structure and negativity in length nerve

(152,

nerves

It

crossed Wal-

of the negative been described

weakening

pheral

with undergoing

comparison

leukodystrophies, 153, 154) and

brain

by nu214). Fig-

Similar fragmentation globules and ovoids

disappearance myehin has

of

described

which

oriented so that in lipid-extracted in paraffin sections, peripheral

as

lerian

nerve fibers. into birefringent

tran-

published 147, 163,

appearance nerve

the

The

by lipids

experimental

been 146,

ure 10 shows the polars of a peripheral

nophilic (42, 115,

birefrin-

after

of nerves have authors (48, 78,

sequent gence

100

of peripheral

is negatively

is caused

38

discussed

sheath

tissue

birefringence

on

known

myehin

nervous

are radially material,

nervous

is presented

Maple

central

negative

the

in the present heading. The of amyloid, is discussed on second, lipid storage diseases

33

PATHOLOGY

of demyelination

section merous

ELEMENTS

are not included first, deposition pages 23-26. The

DIAGNOSTIC

like for

or

Weiss the

demon-

on oxidation groups.

with acid KMnO4, the granules by a pseudocyanin dye. These dyes

of After are can

34

FIG. 9. Myelinated myelin sheaths have R. Schnabel of Jena,

WOLMAN

nerve fibers of peripheral nerve of a rat sectioned the Maltese cross appearance; right, birefringence German Democratic Republic (203).

FIG. 10. Rat sciatic nerve tion 10 days after transection. right, the normal contralateral

teased in Ringer’s solution. The continuity is broken nerve with preservation

Left, fibers and myelin of structure.

transversely is homogeneous.

and

not By

stained. courtesy

of a nerve undergoing Wallerian is transformed into globules and

of

Left, Dr.

degeneraellipsoids;

POLARIZATION

be

shown

hibit

by

their

fluorescence.

metachromasia,

orderly

positively and

of

the

the dye allows their light.

It is known

(195)

that

reaction.

(202)

exhibit

an

of procedures:

staining, staining

gold impregnation, by the Kanzler

violet

or

methyl

dichroitic analyzer polarizer

of

enhancement

was

The

outer

are anisotropic changes when

241).

These

retical

rotating

found patients.

may be substances

usefully which

Organellar specialized

showed

the

and cellular nuclear

under

crossed in

polars.

edge

that

blood, tense

the erythrocyte birefringence.

studied in the

central

were

polarized while the

and

the

birefringent similar

were

radial nature

were

cells

of the

same

not

birefringent.

CELL

These

STRUCTURES

staining

plasma membranes structures: Baud envelope

by

cells with sodium sulfoantimonate. esting that this procedure shows arrangement of protein rather than

staining

is

with

of

been the examina-

prolonged

knowlperipheral

exhibit studied

ghosts birefringence.

and

inthe found

the postsynaptic junction can

microscopy. of the tetanic

changed

be

Changes membrane stimulation

of acetylcholine.

Both

after

denerva-

of

of microfilaments of the mitotic shown by Inou#{233}and Sato (86).

and Follett filaments

structures

(63) showed in cultured seemed

to

of the cells. and Riethmiiller

the granules of rabbits,

of polymorphonuclear believed to represent

birefringent

when

stained

Nucleoproteintaming structures: by polarized light charges

(13)

ing

the

been

movement Missmahl

and

It is interthe ordered lipid moie-

and

alignment

of

shown that myoneural

birefringence

Birefringence spindle has

are highly they are ar-

by aligned.

of erythrocyte mainly form

after

Goldman bundles

birefringence The anisotropy

ordered

membranes Mitchison (131)

administration

of

dur-

birefringence

tion.

SECRETIONS

layers, their for detection.

found

types

were

oxidation

smears

by polarized light intrinsic birefringence

after

KI) and membranes

It is common

air-dried

bodies. (211),

that

with

membrane has studies including

(33) has of the

hepatic

increased become

erythrocyte of numerous

Csillik membrane

spherical Seitelberger

precipitaby potassium

lipids.

process

this

theo-

the

were tolui-

reticulum

the

therefore,

membrane

tion

receptor

destroyed

presumably, The subject

subsequent

Interestingly,

In

Although all cellular membranes ordered structures, except when in multiple too weak

fixation

fixation, either

complex

endoplasmic

was

MEMBRANES,

AND

ranged usually

ing

with

or together arabic. Plasma

the

birefringence that this

quite

and

OF

of

demonstrated.

of great

large and

deposits

BIREFRINGENCE

those

well

to be applicable

contains

These

and

proposed the the birefrinAfter

stained with

(alone gum

(see

parallel in them.

changes

dye-substrate

the

the

were

or rivanol,

the

that

membranes.

seem

reported

of a not

blue of the

in

production

retinal

dine

structures

autooxidative

ferricyanide mounting

to the in the cell

by the

disease:

reviews,

in cardiac

clearly, gentian

cellular

tion

by

De#{225}k(182, 183) for increasing

although do not

part

a red

It is believed

of these bodies gives the typical appearance of’ a spherocrystal, Deposits

after

colors, hut the possibilcannot be ruled out.

pathology. body

peripheral bars.

sections

or Congo

of

by

birefrincells

caused

membranous

avoided,

by

the

of

affected The

absorbing

is probably

Romh#{225}nyi and following procedure

not

solvents.

and

32)

these

enhanced

many neurons contain Seitelberger et al. (212) core light

alignment

is

lipid

secreting

that

Schnabel

of

of

31 and

which

most with

phenomena,

in extensive

pages

and the sign of birefrinthe cells are damaged (172,

importance,

to diagnostic Myoclonus

gence

during

and procedure

caused

segments

cells gence

and orderly with

with

gence

the perpendicular intense colors

of additive polarization ity of a Cotton effect

birefringence

phenol

blue

of a ghioblastoma.

the

pretreatment

length,

dichroism

staining

by 10#{176} from and obtained

processes the

violet.

effect

as

35

PATHOLOGY

by the

showed

trypan

DIAGNOSTIC

ties,

are

to their

intense

number

ex-

processes

affected

IN

of the

groups

astrocyte

is not

Schnabel

processes

SO3H

in respect

positivity

also

because

molecules. This easy demonstration

birefringent

this

They

however,

alignment

consequently alignment polarized

MICROSCOPY

on

and

has been ribonucleic acid hibit

the known acid

(DNA) negative

play (130)

with

a

role

showed

in that

leukocytes lysosomes, are toluidine

blue.

and mucoprotein-conThese can be demonstrated microscopy as the negative

molecules

a tendency

anisotropic fibroblasts.

mostly for

cause

parallel

stretch-

alignment.

for some time that fibrils (RNA) and deoxyribonucleic

prepared or birefringence

stretched (169).

in

It of

vitro exRomh#{225}nyi

36

WOLMAN

and

Jobst

(184)

DNA in microscopy. study

were

tissue The have

Sugar

(230)

drugs

used

pioneers

polarized inherent

in DNA

by

polarization

apparently studied in

been the

the

the

effect

on

cells. The author used (Ehrhich ascites carcinoma

ascites mustard

lymphoma) drug and

These

studies

studying nucleoprotein

and

open

induced

organization

RNA

polarized

up

new

of genetic

material.

study

with

toluidine blue with ferricyanide

tryptic

Many Leme

years (105),

that

the

in

derma cutanea,

could

with crystals.

digestion

of histones,

ary

in re-

or hydrolysis

et al. the

and

were

positively

birefringence

of

mucus

mucus-secreting salivary

tumors.

Alcian

Blue

in addition

same

colon

The

authors

8GS

Alcian in order

in the

ride-containing

granules

DEPOSITION

OF

acid

of

in mixed

used

and

them

a study

and

blue

of birefringence. et al. (70) before

phenomena

in

in the

to toluidine

intensity Gyenge

Musy confirmed

droplets

cells

gland

Blue, the and

birefringent threads. (138)

Alcian

Green

8GX

to increase

These authors observed the

mucopolysaccha-

of mast

BIREFRINGENT

cells. LIPIDS

IN

THE

TISSUES

Crystallization tions ing

of various normal

gent

fat the

indicates acids

adipose

tissue, crystals

droplets. assumption soaps

that

tissues, often (110)

show

secinclud-

birefrinsituated

justly

stressed

crystallization

with is

Frozen

sometimes Lison

breakdown and

lipids:

lipid-containing

acicular

within that

of tissue

completely

of fats

formation

of wrong.

in

body

Scott

operated

165),

Crystallization might be caused

and

to the length of the and Musy and Gotzos

temperature fats.

(28)

fat on

In

observed

subcutaneous

been

(164,

soaps various

asthma

had

esters. It is is caused by

of the

and the

that

of an

under

hypo-

of fats, shown by the and soaps in many

might

represent

a second-

change.

and that this effect was not obtained in a renal infarct. White, Elmes and Walsh (249) found that the thick mucous secretions of chronic bronchitis spect (139)

found of triglycerides

body

The hydrolysis of fatty acids

instances be

Stahlman

tissue

of birefrin-

mainly

point

in

sub-

authors

of the

changes

sclero-

adipose

appearance The

melting

Collins,

thermia. presence

in the

the

lowering

who

called

some cholesterol this crystallization

to the

in

adiponecrosis

consisted

with that

and

separate artifactual

its counterpart

newborns

inflammation

acicular

similar

has

or

associated

relation

author

of

gent

fact,

only

disease

crystals

partly This

Harrison and McNee (73), (217) and others observed

was

excessive

a

ago Siwe

the

of

tripalmitin

to cold

neonatorum

of

staining for

The

nuclei

due

admixed probable

dis-

temperature, crystallize and fluid triolein.

pathology.

infant

of chromatin.

after

the

the

can still

human

these

decreased.

possibilities

associated

interphase

demonstrated

in

after stainthe birefrinwas

processes

light that

acids

in the regular alignment molecules in naturally occurring

Romh#{225}nyi (179) used followed by treatment found

neo-

two types of and NK/LY

and measured treated cells

DNA

changes

crystallization

human

of nucleic

of

miused.

and treated them with a a Vinca alkaloid. The bire-

was studied rivanol. In

of both

and

birefringence

lowering tristearin from the

antimitotic

of

plasms

gence

of

of

light in the

seldom

treatment

cancer tumors

fringence ing with

of study

by

of changes

croscopy

the

sections possibilities

(76,

222,

223).

crystals

a result

agents cells

feature Figure

and

of

enzymic

breaking

down

or the constituent fats (220). The presence of birefringent mainly

known

polarized

acids

of traumatic,

for

of cholesterol

years

as

of this disease process 11 shows the appearance

under

tissues fats

of fats) occurs in with fat necrosis

capable

consisting been

of fatty

of hydrolysis associated

as

other

in the in saturated

Crystallization

(products conditions

adipose tissue Atheromatosis: had

of triglycerides by a diet rich

light.

Also

disease produced in diet rich in cholesterol,,

esters

an

important

(74, 91, of these in the

susceptible deposition

98, 102). crystals

experimental animals by of anisotropic

lipid crystals observers (7).

in the aorta was In some animals

noted by susceptible

atheromatosis, which are

such very

and in animals to this process,

not

such

as dogs

gent

crystals

and

as

rats,

the

occurrence

in experimental

a constant feature (213) considered lipids without

as chicks, susceptible

considered

With

atherosclerosis.

similar

early to

of birefrin-

atheromata

is not

(21, 108, 109). Some authors the presence of birefringent

a distinctive sign of atheromatosis, which the process in the arteries

fatty

a

to

but

not

identical

was with

POLARIZATION

Atheromatous deposit acicular crystals and

11.

FIG.

birefringent

The detection of atheromata acquired atheromatous

had clinical

170). small

The occurrence arteries because

tous

plaques,

a not

of polarized the

by these (65, 72,

on

feasible,

but

by

fundoscopy

deposition ing mainly esters

change

and

may

processes,

in the

lipids

in

histiocytes terolosis, cases

presence

the

eye

milieu

in and (as

for

lipid

normal

example

mucosa of different

the

in cholesorgans in

or without supply

of in

to the

of polarization

The

authors

the

microscopic

studied

type

II

ment)

(53)

intense time the transformed

granules presence mixtures

and

161).

to determine is lost of the

have

in an aniso-

shown

the

that

xanthoma

birefringence at an early lipid droplets are progresinto chromolipid (pigthey

of

160,

of warming

in order

et al.

pri-

an

effect

hyperhipoproteinemia

exhibit With being

basic

changes effective

(156,

the

sections

the

generalized,

microscopy

also

ety of xanthomatous and generalized

concentrations

century

secondary xanthomatous and scientists made

use

in

primary, as in as in diabetic

mary and was available

The glyceride

inflammatory

of hyperlipidemia), of increased

by

consistcholes-

in apparently

of high

of the gallbladder or in histiocytes

foamy

Xanthomatous

chronic

of this localized,

(e.g.,

or disseminated

It may be or secondary,

decade

of intensely

be localized only)

about

cells stage. sively

polarized

droplets especially

Masses

knowledge

in

caused

organs. disease,

hyperlipemia. In the first

may

or spleen

tropic substance. Recently Ferrans

been

denotes cells

skin

aorta.

the point at which birefringence attempt to better define the nature

birefrinIt is obvi-

never

with

term

discrete lipids,

shiny

atheromatous

vivo

triglycerides.

occur

in tumors

tissue

in

This

the

has

histiocytic

of multiple of nonpolar

of

revealed plaques.

of the

of

caused

described et al. (36) of a patient.

apparently

Xanthomatosis:

for

changes

arterioles

to diagnose

the

examination

the

examination retinal bright

transformation

evidence

lesions,

observation

in retinal

accomplished,

some

and

fundoscopic

Postmortem gence of the

terol

light

process

plaques

emboli light.

the

in

of a human section. change

kidney,

in many Wolman’s

(134,

37

PATHOLOGY

Xanthomatous the

recog-

embohi of atheroma-

embohi have been repeatedly 89, 90, 118, 205). David

reported

ously

of

in

entity

DIAGNOSTIC

ester) in the intima Unstained frozen

after

become

of cholesterol of erosion

appearance

diagnosing

yellow

birefringent crystals major importance

uncommon

as

IN

(mainly cholesterol tiny spherocrystals.

embohization

nized

importance

MICROSCOPY

lose

anisotropic has been

the

biref’ringence.

cholesterol-trireported in a vari-

processes: (a) in localized cutaneous xanthomata of dif-

ferent kinds (148, 160, 227, 251); (b) in the Hand-SchUller-Christian disease and in other systemic thomatous (c) in

processes

undergoing

transformation localized chronic

any

xanthomatous

cells.

cerebrotendinous

change

secondary (22, 114, inflammations

(12, 62, 106); (d)

xanthomatosis

(2:39;

xan-

119,

228); with in the

(e)

in

a

38

WOLMAN

group

of primary

xanthomatoses

gier

disease

(11),

and and

cholesterol ester storage (I) in experimental

including

Wolman’s

disease

Tan-

(111,

served

256)

disease (151, xanthomatosis

191); (84,

247).

bone

case

disease,

of this

in

hepatic

respect

in

Also, Seitelberger

the storing processes change, are

presence

cells. In associated

some free

while

or

the

of

tian

disease,

few

anisotropic

cases

Lane

lipids,

For

example,

Smith

(99)

on

histiocytes

droplets

the

in other

droplets

were

In

the

sites

and

to have

presence

of

any

(myehin

structure

is birefringent.

proportion allowing

figures)

of polar lipids fluidity, a similar

ides and cholesterol birefringence except external Figure nia

so In

surface

the

of a low

esters may for the weak

of the

exhibit no effect of the as shown

droplet,

lipid pneumonia

esters group

crystals

found processes

microscopy

can

in greatly

help

the

crystals

in the

storing

is

the

cell a definite

Missmahl

presence

weak presence

and

sign KUbler

of intense

positive

intrinsic birefringence In addition some cells

crystals, presumably the lipid moieties Niemann-Pick

of

to were

on

Maltese

polars

idiocy

in Nie-

(Tay-Sachs

in

the

Tay-Sachs idiocy

granules

noted

been

observation

of

amaurotic

storage

are

already

of dogs

birefringent

by

Diezel

(42)

granules of the late forms are isotropic. This corresponds nature (258), as in most

granules

these pigment numerous mass.

microscopic

Also

the

(19). It has

polymerization

cross-links

produces

lipidosis The

of of

through an

amorphous

(metachromatic metachromatic deposits

found in this disease in the in other organs are intensely

nervous system and birefringent (50). It

is

metachromatic

not

known

whether

deposits

are

possibly

be

Peiffer which

the birefringent,

Dayan

a

of practical

uripoint

which

importance

(37) reported

that

for

the

cresyl violet procedure has been successfully

Hirsch

for sulfaused for

conwith

one

Another light

for example the

lipid

polarization disease

diffusum):

This

finding

of

muscle

and

authors in a

the

against

this

diagno-

establishing

(129)

described and

the

negative

of the fibrils of Gaucher the birefringence found to contain

of the sphero-

caused by separation from the fibrillar proteins. disease: Smetana (219)

207).

(angiokeratoma is

some

crosses

under

crossed

DEPOSITION

OF

SUBSTANCES

which

in cells

has

birefringent

in spherocrystals

in

lipids often

the correct

The

corporis

process

epithelial

sediment

ranged

Urates: associated

a

birefringent

urinary

of oh-

plate.

Fabry’s

that some birefringence

form

giving

diagnosis

exudate,

so

large.

reported

processes

in the pericardium (34, 206, 229). Gaucher’s disease: In this process birefringence considered

in

diagnosing this disease, gives a golden brown polarization color under crossed polars and exhibits distinct dichroism when examined with

(245). polarized

exudative

are

others

structure

that the storage amaurotic idiocy to their chromohipid

and tides,

cholesterol

in a

found

amounts

crossed

electron

(232).

diagnosis.

pneumonia,

under

lamellar

might

pneumoendoge-

of

the lung in which

of inflammatory

cholesterol

to chronic so-called

or cholesterol

birefringent are of

sists

to the the

nary

which

cells. fibrils,

in

were

spherocrystals

Sulfatide leukodystrophy):

12.

nous

sis.

whole

presence

or in other conditions mixture or triglycer-

Reference might be made with lipid crystal deposits,

in

that

the

the

(210) and

of

(G512-type)

ciently high proportion of polar lipids and when the temperature is not high enough to disrupt orderly alignment, the lipids are arranged in lamellae

cells

amaurotic

granules

a suffi-

none

in

nodes

disease): The storing neurons of this disease give intense birefringence (42). This fact correon

very

numerous.

are not believed

presence

sponds

contained

while

Kupffer

Infantile

in

crystals

lymph

and

while

cross appearance mann-Pick neurons.

of Hand-SchUller-Chris-

some

significance.

the

areas

anisotropic

and

cases

differences

deep

or some

of them.

birefringent

These

of

lipids

of the various xant homatous

organs

free full

in four

one with

some

almost are

report

any

cases,

others

findings

of birefringent

marrow

the liver and spleen. According to Diezel (42) the amount of birefringent material in storing neurons in this disease is rather small, while in

The literature dealing with xanthomatous processes is full of apparent inconsistencies to the

of birefringent

large amounts

the thymus,

as

well

been

diagnosis

the

endothelial, as

helpful is

appearing

often as

Maltese

polars.

IN

In gout and with increased

THE

in

NONLIPIDIC TISSUES

other

purine

in

(56, 60, 162,

glycolipid

ENDOGENOUS

in

conditions

base

release,

ar-

POLARIZATION

12.

FIG.

Illustration

of the

MICROSCOPY

difference

between

IN

DIAGNOSTIC

an anisotropic

myelin

figure

urate

crystals

in the Urate lost

may

and

crystals

are

to

or

birefringence

and

in

by

in respect

are and

to length

oxalosis

tions

associated

preserved positive

of the

crystals

use

of’ a sim-

Waldmann

(246)

suggested

the

ple

inverted

polarizing

microscope

ing

various

crystalline

deposits

tions. As this temperatures, of

instrument the author

temperature,

reagents

on

urates,

oxalates

pounds.

Sorger

mended

the

techniques

the

and

use

and

various Bausch

tissue

and

various

thus

identify

inorganic (221) also

of’ crystallographic identifying

sec-

the use of high determine the

solvents

and

for

in

allows could

crystals

identifv-

for

comrecom-

polarization urinary

sediment

crystals. Oxalates:

in the

urinary

These tract

compounds can and occasionally

in a number of’ conditions. their birefringence in the

They kidneys

high

chloride

doses

of’ ammonium

be found elsewhere

were f’ound by of’ rats given and

oxalate

158)

with new

oxalate: of sodium brownish yellow

under

patients.

(Fig.

15).

Roulet

and

crystals

are

author ing.

be (185) out

review

Eberlein

appear-

the

alcohol in

(49)

ease of’

crysorgans

extensive

fixation

sharply

with

tissues.

and In

separated

the

which

stain-

in a

the The

no

crystals

tissues.

de-

and

the

cystine

ocular

incomFigure

in various an

in of’

epiphyseal

changes

out

found

the

gave

pointed

(24) also

the

material. biref’ringent

found

pathologic

dissolved

patients

by a which appear

Macaluso

13 shows

within

the

advocated Bfirki

(24:3

identifying

polars.

Figure

can

of

crystals

condi-

this procedure useful oxalate in autopsies

crystals

of cystine

and

Voigt

oxalate crystals under polars in the kidney, and

such

scription

various for

cartilage in human autopsy Cystinosis: In this disease tals

glycol

(14).

crossed

nephrotic

shows

in

or with

of endoge-

the sections are treated naphthalhydroxamate yellow crystals which

(113) found calcium

14

and

uremia

E,

of both types hydrophilic

in cases

procedure

and Berg identifying

ance of calcium pletely crossed

and

by ethylene

(1, 259),

(100, a

calcium solution produces

molecules lipids)

poisoned

poisons

introduced

bright

be shown.

ef’fects

in humans

nous

the

droplet.

right, the the nonpolar

mostly of

lipid

layer; and

other

aqueous

isotropic

layer

some

because

crystals

formalin

the thin surface of the polar lipids

(46),

an

lipids such as phospholipids arranged in haphazard fashion

tissues.

are

treated

probably urate

fixed

of rods

articulations,

in other

soluble

highly fixed

proteins.

tissues

weak effect layers (the polar lipids).

in some

Occasionally,

binding

the

occasionally

material

reagents.

can

appear

kidneys in

in

except for hydrophobic bulbs of the

and

#{149}, polar

molecules of nonpolar lipids such as cholesterol ester or triglyceride; fatty acids. Left, a lipid droplet, all the molecules except the surface consequent isotropy arranged in alternating layers (between the

39

PATHOLOGY

such

critical cystino-

40

WOLMAN

FIG.

Crystals

13.

incompletely

of calcium

crossed

polars.

in the kidney of Dr. E. Liban

oxalate

(Courtesy

of a patient suffering from endogenous oxalosis of the Beilinson Hospital, Petah Tiqvah.)

_y;:

-

FIG.

Crystals

14.

(Courtesy

sis

of Dr.

from

cystinuria

of’ polarized

light

recognizing

the

again but

of calcium E.

described in the

case

the detection polarized light

oxalate

Liban

of the

and

stressed

the

crystals

of ocular

within

Beilinson

the

epiphyseal

Hospital,

the

microscopy crystals.

Petah

importance

cartilage

amino

acid

and

and

and

(59)

birefringent

Frazier in the

Wong

ocular

cystinosis

no

of’ these crystals seems to have been

in

tissues study

vivo

perfurmed

Hooft

with to

disease:

In

this

disease

of’

metabolism, et

al.

could

or in sections terial.

The

amino

acid,

in aqueous syrup

found

same

(79)

patient

Diezel reported

crystals

crystals of

of the

as in Figure

11.

Tiqvah.)

for visualizing

date. Maple

under

in

in

and on

Martin

the

various

organs.

be found

in fresh-frozen

prepared

from

sections

material

seemed

peptide

or protein

which

glial

cells

Birefringent and

interstitially

of The

ethanol-fixed

crystalline fixatives.

(44)

presence

mato he an is soluble

crystals

were in

the

POLARIZATION

brain,

in liver

Bowman’s

cells,

in tubular

bone within

by

these

marrow epithelia

and and

in many in stromal

other and

needles

any

when

tissues

to aqueous

liquid

be

of

are examined

cut

in

crossed

of

cryostat in the

pyrophosphate

(pseudogout):

often

(or

by

by

eventually

polarized

hibit

aceous sign

tanning

light

weakly

urates

(155,

synovial 218).

positive

and

appearance

The

synovia

of

membrane)

echinococcus

and

ex-

cyst

Crystalline shown by

Figure

16 shows

in such

a case.

the

OF

BIREFRINGENT

Formalin

FIG.

crossed

BODIES

pigment:

15. Cystine polars.

EXOGENOUS

crystals

(Courtesy

NONLIPIDIC IN

THE

can

and

even

is

This

in the of Dr.

E.

in

reality

spleen

of a cystinosis

Liban

of the

Beilinson

of sign

effect

of’ the

Ebner’s

reac-

birefringence

from

the

sections

of

the

lumen

of’ a

scolices

sediment

of’

of the

cation

patient. Hospital.

of

can example,

This

be by

methodology

identifying

them

(246).

This

nodes

(58).

fur positively condition

examination

lymph an

of von

the

Pindborg

be used

tissue

the

TISSUES

change the

drugs: Sulfonamides polarized light, as, for

Silicosis: PRESENCE

to that

results into

fluid.

Fransden in

changes maturation.

The and

p

protein-

of the proteins which changes

tanning

ex-

(107,

the

of an oxyuris lying in the appendix. Figure 18 shows

the

polars of a wet them from crystals

that

during

obtained

and

arthropods

maturation.

other For

cellulose

out

of some

on collagen. Figure 17 shows

of’ synovial crystals

pointed

is similar

and

helminths

and

birefringence

with

cuticle anisotropy.

199)

to quinone

cuticle human

hirefringence

cholesterol. of the

deposi-

examination of the

exhibit

cuticle

in

p :391). fungi.

The lamellar orientation in positive birefringence

some

is

various

fixed

as birefringent

The

(196.

outer

in

which

(107,

503) are birefringent. Schmidt (197)

is due

is a condition

examination under crossed smear allows to differentiate of

chitin

negativity

and

dihydrate

This

diagnosed

fluid

ample,

organs

appears

organisms: of

41

blood-rich

or rhomboids

ectoparasites

tion

Calcium

pigment

some

state

instances. tion

in

This

layers

dry

importance

formed

outer

the

withStudy

the

polars

diagnostic

of

or preferably

whatsoever.

sections under

in some

reagents,

reagents

fresh-frozen examined might

microscopy

PATHOLOGY

Parasitic

biopsy-based studies were that crystals might be

polarization

exposure

to

artifact formalin.

of amino acid metabolism: processes only a limited num-

processes

out

in

adre-

autopsyor It is possible

detected

and

in the

cells.

diseases of these

ber of made.

cells

DIAGNOSTIC

kidneys,

reticuloendothehial Other In some

lining

IN

in the

capsule

nal medulla, organs both

MICROSCOPY

of sections under

the

Unstained Petah

crossed

birefringent

cryostat Tiqvah).

can

be diagnosed

of af’fected polars

(64).

material

section

examined

by

lungs

or

Identifias

silica

under

42

WOLMA,N

FIG.

cryostat

16. Calcium pyrophosphate section under crossed

dihydrate

crystals

within

the

synovia

of a pseudogout

patient.

Unstained

polars.

with

tissue

tropic

constituents

(18).

use

phase

their

of’ten

authorities

microscopy

fibers (8). importance. for

and

Some

f’or

iso-

in fact,

identifying

to

asbestos

Study under crossed polars is of’ however, in the screening of’ dusts

asbestos

content

Extraneous

(140).

artificially

materials:

introduced

Experimental

human

become

prefer.

beings

animals

and

are of’ten injected

also

with solutions

containing formed impurities. These f’ormed contaminants may cause clinical or subclinical embohization and in many instances they can be detected by polarized light microscopy. Innes, Donati

and

mice

hirefringent

their

detection.

17.

Oxyuris The

depends

on

procedure

about crystals

to

hr

be presumed

fibers, after

The be made to polarized

as some

such, time

exhibit in the

for

Birefringent

incineration

may

of’ asbestosis

tissue

light

sections

microscopy. weak

body

birefringence they

become

can without

Asbestos and coated

and this Some

with

of’ injection,

has

injecting

themselves

phenidate

tablets

of other

drugs

retinopathy

methylphenidate one patient was

characteristic

inject

tablets

intended in

and found

such

as pieces

themfor

water.

described

The

80)

10:3). in

in patients

dissolved and

methyl-

possibly

Atlee

(9)

patients

at autopsy to contain

In firearm wounds and in stances of trauma, hirefringent terial,

in obvi-

helped

addicts

with

(47,

was

starch embohi in the lung organs, as well as at the been

(71,

embohi

hair

suspended

occurrence of talc and and in parenchymatous site

hair

The

drug

administration

peculiar

of SiO2.

diagnosis on

used

sections

(67).

the

to consist

of an

commonly

paraffin

550-600#{176}C

lumen

is birefringent.

The

withstand

Asbestosis:

the

worm

incinerate at

which

usually recourse

within

of the

pretreatment. is

2-3

worm

cuticle

fuund

tests.

intravenously

oral FIG.

(85)

in toxicity

ously selves

appendix.

Yevich

used

tablets

described

the starch

rock,

to

retina emboli.

various other crystalline

of’ calcareous

a

addicted

of’ inma-

pieces

POLARIZATION

FIG.

Scolices

18.

inner

structures

of’clothing

and

intensely

vegetal

the tissue. Polarized in detecting For

these,

example,

on a which

of’

these

Both

fur

talcum

exposed

stasis

and

other

sis

of’ regional

may

on

microscopic

In conclusion, polarized

chronic

4.

of’ grave

import

hut

been

help

a

the

use

few

of’

pathologists

substances which might have in ordinary light microscopic

escaped exami-

nation

and

physical

establish

the

nature

of’ some

anisGtropic

posits.

The

of’

unfixed

use

cryostat

chemical structures

polarization

sections

and

and

help

Ambronn

Uber

H:

on

detect

in some

is further might

cuticle

and

meta-

suggested be

useful

in

measuring hirefringence

constituents. das

6.

Surgi1951,

film

phase retardain fibrillar

Exp

Cell

Zusammenwirken

und

or

Res von

Eigendoppelbre-

chung I. Kolloid Z 18:90, 1916 Angmar B: Studies on the ultrastructure of dental enamel. VII. A microradiographic and polarization microscopic study on developing

enamel.

Odontol

Revv

16:167,

1965

Angmar B, Carlstr#{246}m D, Glas JE: Studies on the ultrastructure of dental enamel. 3 Ultrastruct Res 8:12, 1963 Angmar-Mansson B: Studies on the distribution and ultrastructure of the main components in

human holm,

dental

enamel.

Doctoral

thesis,

Stock-

1970

Kaninchenaorta terinsteatose.

de-

microscopy

might

cell

7. Anitschkow

detect detection

to

method weak

membranous 37:230, 1965

human 5.

polarization

that

can

made

of’

outer

examinations.

Stabchendoppelhrechung

a piece of’ diagno-

have

It

optics

densitometric tions due

lymph

presence operation.

microscopy

Tears

tissues.

examined a tentative

crystals

processes.

f’undoscopic

The

LITERATURE CITED AC (Ed): The Kidney: Medical and cal Diseases. Grune & Stratton, New \ork, p 228 2. Allen RD, Nakajima H: Two-exposure,

adhesions,

it is suggested

light

polarization

3.

would the a past

that

fureign dealing

examination,

microscopy revealed talcum grains f’rom

disease

spillage

produce

fluid.

of water-soluble

bolic

of’ten with operations

complications

I have recently a patient where

from the sediment of cyst under crossed polars.

deposits

43

PATHOLOGY

1. Allen

commonly

patients’

surfaces,

enteritis

in

gloves.

DIAGNOSTIC

ocular

bodies

cause

the

starch

serosal

may ensue. bowel f’rom

are

lubricating

reactions, cells. In surgical

with

reported

conjunctivitis foreign

starch

into

and

granulomatous body giant

(257)

occasionally

substances

in

useful

microscopy.

and

by surgeons other accidents

is

materials.

Tibble

standing threadlike

powder

can be found

unstained,

and

by polarization

detected

used and

often

Wolter

Talcum

matter,

IN

obtained Wet smear

light microscopy

case of’ long birefringent

were

echinococcus birefringent.

of Taenia

are

MICROSCOPY

N:

(,Jber die Ver#{228}nderungen der bei experi menteller C holesBeitr Pathol 56:379, 1913

8. Ashcrof’t T, Heppleston AG: The optical and electron microscopic determination of pulmonary asbestos fibre concentration and its relation to the human pathological reaction. J Clin Pathol 26:224, 197:3

44

WOLMAN

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28.

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MICROSCOPY

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