Multimodal Laboratory of Cell Adhesion and Motility
Nencki Institute of Experimental Biology
Polish Academy of Sciences
Pasteura 3 St., 02-093 Warsaw
tel. +48 22 589 2471; e-mail: email@example.com
Head: Paweł Pomorski
The Multimodal Laboratory for Cell Adhesion and Motility Analysis was opened in Nencki Institute of Experimental Biology PAS as a part of NanoFun project. It is a specialized workstation enabling analysis of nonsynchronous processes on single cell level. It is equipped with epifluorescence microscope Leica AF 7000 designed for long-term in vivo experiments. The system utilizes noninvasive light microscopy to trace cells adhesion and trajectory of movement. This includes long term time-laps microscopy using transmitted light techniques like DIC as well as fluorescence markers.
For data analysis specialized software is available. The automation of the microscope stage together with controlling software allow to design multimodal, long term experiments in multiwell plates or multichannel systems for chemotaxis analysis. Cell adhesion can be analyzed in vivo with IRM (Interference Reflection Microscopy), a method which enables to visualize regions of cell adhesion to the surface. Microscope offers the possibility to conduct various experiments utilizing wide-field fluorescence microscopy, moreover fast optics (fast filter wheels) of the microscope makes it particularly suitable for experiments using fluorescent molecular probes like calcium probe Fura 2.
Current experiments involve time-lapse registration of cell trajectories in twelve and twenty-four well plates with registration of at least three fields of view per well. It allow the researcher to test multiple experimental setups on exactly same cell batch. Our laboratory developed own software to correct imperfections in the microscope stage movements. Software for image tiling was written for imaging of whole area of chemotactic experimental chambers, what not only extends imaged area, but also allows researcher to follow cells crossing boundaries of individual camera fields of view.
All software is available both as a Matlab script and an ImageJ plugin. Laboratory owns also software and hardware (low diameter filter wheels) needed for fast imaging with use of molecular probes. It allows to obtain up to 20 pairs of ratiometric Fura 2 measurements per second. Workstation used to control an image acquisition is powerful enough to perform ratiometric calculations on the flight and display live measurement result.