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Activity Report 2010 - CNRS

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7 – NANO APPROACHES<br />

TO LIFE SCIENCES<br />

Life sciences are one of the main<br />

application fields of nanosciences, as<br />

evidenced by the increasing number of<br />

proposals received along the successive<br />

call for proposals. Biological cells and<br />

molecules can be manipulated and<br />

analyzed with the greatest specificity in<br />

small volumes. Innovative devices will<br />

help building tomorrow’s medicine.<br />

MICRO- AND NANO<br />

FABRICATION FOR THE<br />

LIFE SCIENCES<br />

Contribution of 3D microenvironment<br />

to cell<br />

adhesion<br />

“New comers” Project 2008: Martial<br />

BALLAND (LIPhy)<br />

PhD student: Kalpana MANDAL<br />

Biological tissues are complex composite<br />

materials made of cells and intercellular<br />

matrix molecules secreted by the cells.<br />

Their formation and maintenance depend<br />

on both chemical and mechanical cues<br />

present in the microenvironment of each<br />

cell. Tissue biology involves therefore<br />

complex feedback loops. The aim of this<br />

project is to reproduce the organized<br />

geometry of biological tissues and to<br />

analyse quantitatively the mechanical<br />

forces developed at the cell/substrate<br />

and cell/cell interfaces.<br />

In order to measure mechanical forces,<br />

thin hydrogels are prepared that contain<br />

a homogeneous distribution of<br />

fluorescent 200nm beads (Fig. 1). The<br />

hydrogel surface is coated with<br />

fluorescent adhesion proteins, using an<br />

innovative deep-UV irradiation method<br />

developed in collaboration with Manuel<br />

Thery (IRTSV).<br />

This setup paves the way to a functional<br />

analysis of tumor cell invasiveness, which<br />

will be very useful to analyze biopsies<br />

and thus help cancer therapy.<br />

Fig. 1: Top: pictures of the adhesive protein<br />

micropattern (left) and the nanobeads<br />

distribution (right) used to calculate the cellsubstrate<br />

force distribution.<br />

Bottom: distribution of actin microfilaments<br />

(left) and traction forces (right) in a single<br />

micropatterned cell. Barscale: 20 µm<br />

Nanodroplet chip for<br />

controlled assembly of lipid<br />

bilayers and electrical<br />

detection of single-protein<br />

activity<br />

RTRA Project 2008: “Nanobiodrop"<br />

Benjamin CROSS (LEGI)<br />

Post-doctoral fellow: Anne MARTEL (IBS<br />

& LEGI)<br />

Transmembrane channels form a large<br />

class of molecules that play essential<br />

roles in cell physiology by allowing polar<br />

molecules, for instance ions, to cross<br />

biological lipid bilayer membranes. They<br />

are the target of numerous drugs and<br />

toxins. Nevertheless, conventional<br />

electrophysiological methods used to<br />

study ion channel activity are laborious<br />

and slow.<br />

FURTHER READING:<br />

SCIENTIFIC REPORT<br />

Front Biosci (Elite Ed). Jan 1;3:476-88 (2011)<br />

Multi-confocal fluorescence correlation<br />

spectroscopy<br />

The traction force distribution is<br />

computed from the bead displacement<br />

map by a Fast Fourier Traction Cytometry<br />

software. The forces developed by the<br />

micropatterned cells can thus be<br />

calculated and averaged, taking<br />

advantage of the similar geometry of the<br />

patterned cells. Significant differences<br />

between tumor cells have been<br />

evidenced, that are modulated by<br />

different tumorigenic signals.<br />

This innovative project called<br />

‘Nanobiodrop’ uses digital microfluidics to<br />

reconstitute lipid bilayers from two<br />

nanodroplets covered with phospholipids.<br />

Electrowetting is used to manipulate<br />

these droplets and put them into contact.<br />

Some of the nanodroplets are filled with<br />

membrane proteins, which are able to<br />

spontaneously insert in the membrane,<br />

once it is formed. The electrodes are then<br />

used to monitor the ionic current that<br />

flows from one droplet into the apposed<br />

one through the incorporated<br />

transmembrane channels.<br />

CONTACTS<br />

Franz BRUCKERT<br />

franz.bruckert@inpg.fr<br />

Tel: +33 4 56 52 93 21<br />

Julian GARCIA<br />

julian.garcia@ujf-grenoble.fr<br />

Tel: +33 4 56 52 08 31<br />

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