Z-Stack Through a Spheroid Before and After Clearing 

A short video of a z-stack going through a stained L929 spheroid (red: phalloidin, cyan: DAPI) before and after clearing.
(Left) Spheroid before clearing. Note that due to light scattering and absorption only peripheral cells are visible, whereas the cells in the center cannot be seen. (Right) Spheroid after clearing. Clearing of the spheroids allows the visualization of cells even at the center of the spheroid fibroblasts while at the same time preserving the spheroid`s morphology. Counting of cell nuclei would even allow the determination of cell numbers forming this spheroid. Confocal microscopy, z-stack of 440 slices at a slice spacing of 0.36 µm.
The spheroid was created and cultured in the µ-Slide Spheroid Perfusion, the images were taken after 7 days of perfusion with the ibidi Pump System, 0.74 ml/min.
Julian Hofmann, Selina J. Keppler
Inflammation and Immunity Lab, TranslaTUM, Institute for Clinical Chemistry and Pathobiochemistry, Klinikum rechts der Isar, Technical University Munich, Germany.
More details in our User Protocol 11: Spheroid Culture, Staining, and Clearing for 3D Imaging
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