Organelles positioning assay

4DCELL DEVICE

Micropatterns

 

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).