Hi everyone! I want to perform an immunofluorescent staining of spheroids from human glioblastoma cells fixed in 4% PFA. I found many protocols and papers suggesting different solutions and times for permeabilization, blocking, and incubation with the primary and secondary antibodies.
Can anyone suggest a suitable protocol to perform a good immunofluorescent staining of whole spheroids?
Hi Allesandra,
I can highly suggest this protocol. It was elaborated for whole organoids (ca. 500µm): Fluorescence-based Single-cell Analysis of Whole-mount-stained and Cleared Microtissues and Organoids for High Throughput Screening (bio-protocol.org)
It worked really well for our organoids (diameter 500-1000µm). How big are your GBM-spheroids?
Best,
Nicole
Nicole Christin Riedel my spheroids have a diameter of less than 500μm. Anyway, thanks for your suggestion, I'll see if this protocol may be appropriate.
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