Institute of Automation and Control

Microrobotics for Cell Manipulation: from Human Cells to Wood Cells? COST E54, Opening Seminar, Riga Micro- and Nanosystems Research Group Dr. Pasi Kallio 25.4.2007

COSTE54 Opening Seminar, Riga, April 2007

Speaker Introduction • Senior Research Scientist at TUT / Institute of Automation and Control • Head of a Micro and Nanosystems Research Group • Research on microrobotic manipulation since 1995 – Microrobotic manipulation of human cells – Handling and assembly of mechanical and optical microcomponents TAMPERE UNIVERSITY OF TECHNOLOGY Institute of Automation and Control

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COSTE54 Opening Seminar, Riga, April 2007

Micro- and Nanosystems Research Group • Expertise – – – –

Microrobotics Microfluidics Active actuator materials Automation and control

• Application areas – Cell and molecular biology: automation of the research tools – Biomedical diagnostics: automation and miniaturisation

• Personnel – 15 researchers and reasearch assistants

TAMPERE UNIVERSITY OF TECHNOLOGY Institute of Automation and Control

COSTE54 Opening Seminar, Riga, April 2007

Outline • Background – What is micromanipulation – Application areas – Cell biology application

• Analysis – Functional end-user requirements – Performance requirements

• Examples of Research at TUT TAMPERE UNIVERSITY OF TECHNOLOGY Institute of Automation and Control

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COSTE54 Opening Seminar, Riga, April 2007

Micro- and Nanomanipulation Strategies Micromanipulation strategies

Contact methods

• Micromanipulation – Handling of objects whose dimensions are appr. 1 µm … 1 mm

• Nanomanipulation

Non-contact methods Magnetic Optic Electric Acoustic

– Handling of objects whose dimensions are smaller than 1 µm TAMPERE UNIVERSITY OF TECHNOLOGY Institute of Automation and Control

COSTE54 Opening Seminar, Riga, April 2007

Micro- and Nanomanipulation • Manipulation of targets ranging from sub-µm to a few mm in size • Applications – Cell biology – Microassembly – Material Science

TAMPERE UNIVERSITY OF TECHNOLOGY Institute of Automation and Control

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COSTE54 Opening Seminar, Riga, April 2007

Micromanipulation in Cell Biology Biomicromanipulation

Eukaryotic Cells

Adherent

Non-contact

Tissues

Prokaryotic Cells

DNA, Protein Crystals

Suspended

Contact

Non-contact

Contact

Microinjection

Aspiration

Measurements

TAMPERE UNIVERSITY OF TECHNOLOGY Institute of Automation and Control

COSTE54 Opening Seminar, Riga, April 2007

Adherent vs. Suspended Human Cells

• Adherent cells

• Suspended cells

– Size a few tens of µm in diameter – Liver cells – Ephithelial cells – Neuronal cells – ...

– Germ cells (egg cells 100 µm) – Blood cells (10 µm)

Neuronal Cells

TAMPERE UNIVERSITY OF TECHNOLOGY Institute of Automation and Control

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COSTE54 Opening Seminar, Riga, April 2007

Application Case: In-vitro Toxicology • To use in-vitro cell cultures to study toxicological effects of chemical compounds – such as drugs, cosmetics, heavy metals,…

• Scientifically reliable, economic, efficient and ethical replacement of – Laboratory animals – Human experiment (i.e. the real process)

• Used for academic and industrial R&D

COSTE54 Opening Seminar, Riga, April 2007

TAMPERE UNIVERSITY OF TECHNOLOGY Institute of Automation and Control

? TAMPERE UNIVERSITY OF TECHNOLOGY Institute of Automation and Control

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COSTE54 Opening Seminar, Riga, April 2007

Examples of Intracellular Microinjections

FITC

GFP

Available at http://www.ac.tut.fi/aci/mst (VIDEOS) TAMPERE UNIVERSITY OF TECHNOLOGY Institute of Automation and Control

COSTE54 Opening Seminar, Riga, April 2007

History • Inception of cell injections already as early as 1911 • The use of micropipettes as intracellular electrodes in 1946 • Injections of mammalian cells of somatic origin in 1970’s • IVF technology in the end of 1970’s • Few citations in 1970’s, thousands in 1990’s • Getting more and more popular due to developments in stem cell and other primary cell cultivations TAMPERE UNIVERSITY OF TECHNOLOGY Institute of Automation and Control

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COSTE54 Opening Seminar, Riga, April 2007

History - Devices • Firstly manual • Use of motors in 1960’s and piezoactuators 1980’s • Current state-of-the-art – Computer-controlled semi-automatic devices

• Currently under development – Increased amount of information using various sensors – Full automation

TAMPERE UNIVERSITY OF TECHNOLOGY Institute of Automation and Control

COSTE54 Opening Seminar, Riga, April 2007

Outline • Background – What is micromanipulation – Application areas – Cell biology application

• Analysis – Functional end-user requirements – Performance requirements

• Examples of Research at TUT TAMPERE UNIVERSITY OF TECHNOLOGY Institute of Automation and Control

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COSTE54 Opening Seminar, Riga, April 2007

End-user Requirements - Functional • Treatment of cells – Mechanical, electrical and biochemical stimulation of cells

• Characterization of cells – Measurement of mechanical, electrical and biochemical properties of individual cells

• Studies of cell interactions • Environment – Providing proper environment conditions is essential for cells to live TAMPERE UNIVERSITY OF TECHNOLOGY Institute of Automation and Control

COSTE54 Opening Seminar, Riga, April 2007

End-user Requirements - Performance • More precise / accurate movements – Intracellular targeting

• More precise / accurate injection volumes – Intracellular targeting – Smaller variation in operations

• Increased throughput – E.g. 10 cells / min

• Less routine manual manipulations • Less special-trained man-power • Studies at the individual cell level

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Automation

– Resolution of a single cell in a cell population, – More precise characterization than studying the population as bulk

• Increasing number of parameters to be measured – E.g. biochemical parameters but also physical parameters such as forces TAMPERE UNIVERSITY OF TECHNOLOGY Institute of Automation and Control

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COSTE54 Opening Seminar, Riga, April 2007

Cellular Stimuli and Responses NUTRIENTS

GROWTH PRODUCTION REPRODUCTION

OTHER CELLS TESTED VARIABLE

CELL

INPUTS ≈ STIMULI

OUTPUTS ≈ REACTION

Electrical (other cells)

Electrical (intercellular) Physical (p – swollen cell)

Physical (T, p, attach) Chemical (nutrients)

Chemical (c)

INTEREST More quantitative measurement data!!

TAMPERE UNIVERSITY OF TECHNOLOGY Institute of Automation and Control

COSTE54 Opening Seminar, Riga, April 2007

Outline • Background – What is micromanipulation – Application areas – Cell biology application

• Analysis – Functional end-user requirements – Performance requirements

• Examples of Research at TUT TAMPERE UNIVERSITY OF TECHNOLOGY Institute of Automation and Control

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Cell Manipulation System COSTE54 Opening Seminar, Riga, April 2007

Current Set-up • Two micromanipulators • Multi-micromanipulator stage for 4 micromanipulators • Injector / aspirator • Contact detection • xy stage • Microscope and CCD camera + fluorescence option • Environment control • Control software TAMPERE UNIVERSITY OF TECHNOLOGY Institute of Automation and Control

COSTE54 Opening Seminar, Riga, April 2007

Characterization of Human Cells

F?

TAMPERE UNIVERSITY OF TECHNOLOGY Institute of Automation and Control

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COSTE54 Opening Seminar, Riga, April 2007

Functionalization of Human Cells • Injection of androgen receptor siRNA molecules into single human prostate cells (LNCap) • Injection of plasmid DNA containing a gene of interest into single cells of a selected cell line • Injection of mRNA coding nef protein into single cells of a selected cell line

TAMPERE UNIVERSITY OF TECHNOLOGY Institute of Automation and Control

COSTE54 Opening Seminar, Riga, April 2007

Mechanical Treatment of Human Cell Populations

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COSTE54 Opening Seminar, Riga, April 2007

Possible Operations for Wood Cells • Fibre treatment – Grinding and fibrillation of individual fibres (mechanical treatments) – Wetting, drying and colouring of single fibres (chemical treatments)

• Fibre characterisation – Mechanical characterization (e.g. tensile strength) – Characterisation of liquid-fibre interactions on single fibre level (including water, dyes as well as other chemical additives) – Study of dislocations and other weak points in single fibres

• Bond treatments – Creation of fibre bonds in a controlled manner.

• Bond characterisation – Characterisation of mechanical properties of a single bond (e.g. bond strength of) TAMPERE UNIVERSITY OF TECHNOLOGY Institute of Automation and Control

COSTE54 Opening Seminar, Riga, April 2007

Wood Cells: Experiments on Mechanical and Chemical Treatment

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COSTE54 Opening Seminar, Riga, April 2007

Conclusions • Treatment and characterization of individual wood cells using micro- and nanorobotic technologies is feasible • Increased and more precise understanding on wood cells and on the effects of various treatments can be expected • Modifications and adjustments to the systems developed for cell biology, microassembly and material science will be needed TAMPERE UNIVERSITY OF TECHNOLOGY Institute of Automation and Control

COSTE54 Opening Seminar, Riga, April 2007

Acknowledgements • Collaborators – – – – – – – – – –

• Research Group at TUT

CEO Juha Korpinen, CMT – Johana Kuncová-Kallio, Professor Timo Ylikomi, UTA – Katrin Kovanen, Professor Hanna Tähti, UTA – Mikko Lukkari, Professor Jouko Viitanen, VTT – Pekka Ronkanen Dr. Heli Skottman, UTA Professor Markku Kulomaa, – Juha Hirvonen UTA – Et al Professor Jukka Lekkala, TUT • Funding Professor Jari Hyttinen, TUT Professor Minna Kellomäki, – Tekes TUT – The Academy of Finland Dr. Marja-Leena Linne, TUT – Several Finnish companies

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COSTE54 Opening Seminar, Riga, April 2007

THANK YOU

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