Speaker
Description
Immunofluorescence is a powerful technique for the detection of multiple specific markers in cells; however, the fixation process prevents the study of the cells’ motility. We therefore propose a tool that helps find back tracked cells after immunofluorescence (IF). A slice of tissue is imaged during 48h at different positions with a 20x objective, creating movies on which cells are manually tracked. For each position, an additional image is captured, using transmitted light microscopy and a 4x objective, for a bigger field of view. Based on the stage displacements, a reconstruction is made with these images and the outlines of the whole biological structures are drawn by the user. Just after the movie, slices are immunostained to determine the cells’ fate and imaged under the microscope. An automatic segmentation of the structures is performed on these images. The tool developed in Fiji/MATLAB is aligning the contours obtained on two paired images and automatically determines the (similarity) transformation between the images. The 20x acquisition areas are displayed on the IF image, aiding in identifying the fate of each tracked cell.
| Authors | Anne-Sophie MACE*, Clarisse Brunet Avalos, Laure Coquand, Alexandre Baffet |
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| Keywords | correlative microscopy, contour-alignment, fluorescence and transmitted light microscopy, ImageJ, multi-stage acquisition |
