Organelles positioning assay
Precise statistics on organelle positioning in the cell
4DCELL DEVICE
READ-OUTS
Quantification of organelles positioning, cell shape standardization
STANDARD CULTURE LIMITATION
In standard cultures, cells are distributed in 2D monolayers causing them to acquire random shapes. This affects the distribution of their internal organelles, precluding to determine their exact positioning in a reproducible way.
NEURONAL NETWORK IN A WELL
Using 4Dcell micropatterns, cells acquire a standard and reproducible shape. This enables the acquisition of high-resolution images, from which precise statistical models on the exact positioning of the organelles can be derived.
EXAMPLE
Cytoskeleton organization of a cell cultivated on triangle micropatterns [1].
Distribution of MTs, F-actin bundles and focal adhesions in triangular cells cultured on triangle micropatterns.
(A, left) Reconstruction of representative MT growth trajectories by superposition of multiple (∼50) consecutive frames (total time, 150 sec). (A, right) FAs marked by vinculin are located exclusively at the vertices of the triangular cells plated on fibronectin.
(B, left) F-actin and (B, middle) focal adhesions were visualized by staining with fluorescent phalloidin or immunostaining with vinculin antibody, respectively.
(B, right) The entire organization is visualized more precisely by supperposition of 25 images (pseudocolored heatmap).