Team SiedyZsig

Imaging a fixed, large spheroid (&oslash; &asymp; 500 µm) on Zeiss Lightsheet Z.1
We used a cell line expressing GFP-tagged histone 2B. The spheroid was embedded together with beads in 1.5 % agarose in a glass capillary (black label).
 * Sample preparation


 * Imaging
 * 1 timepoint, 6 angles, dual-side illumination
 * excitation: 488 nm
 * detection: 20x ...

After approximately 2 h 6 min (2 h to set up the acquisition, 6 min to run it) we had a nice 36 GB data set. Post-processing in progress, images will follow...
 * Results

Outlook: Overnight time-lapse imaging of living, 4-days-old spheroids (&oslash; &asymp; 50 µm) on Zeiss Lightsheet Z.1
H2B-GFP cells were additionally labelled with the live-dye SiR-tubulin (Lukinavičius et al. 2014). They were embedded as single cells in Matrigel and injected into chambers made of either 1 % Gelrite or 1.5 % agarose. We incubated the chambers, and for the past four days the cells have been proliferating and forming small spheroids.
 * Sample preparation


 * Imaging

TBD


 * Results

TBD

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