Fluorescent staining and microscopy

JH Jörn Hülsmann
TF Theresa Fraune
BD Baratha Dodawatta
FR Fabian Reuter
MB Martin Beutner
VB Viktoria Beck
MH Matthias Hackert-Oschätzchen
CO Claus Dieter Ohl
KB Katja Bettenbrock
GJ Gabor Janiga
JW Jens Wippermann
MW Max Wacker
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After stopping cultivation, tubes were washed in cold phosphate buffered saline and fixed in 4% buffered paraformaldehyde and 10% sucrose for 10 minutes. Fluorescent staining of nuclei and cell bodies was realized using combined DNA and RNA staining by acridine orange (AO) according to the manufacture’s recommendations (cat.nr.:10127-02-3, Thermofischer). Fluorescent staining of the cytoskeleton was performed using phalloidin rhodamine (abcam), specifically interacting with F-Actin according to the manufacturer’s recommendations. Microscopic evaluation was done using an Evos FL Cell Imaging System (Thermofischer).

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