PLoS ONE (Jan 2023)

Immunostaining protocol for infiltrating brain cancer spheroids for light-sheet imaging.

  • Benedicte Bjørknes,
  • Oliver Emil Neye,
  • Petra Hamerlik,
  • Liselotte Jauffred

DOI
https://doi.org/10.1371/journal.pone.0281161
Journal volume & issue
Vol. 18, no. 2
p. e0281161

Abstract

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Glioblastoma tumors form in brains' white matter and are fast-growing and aggressive. Poor prognosis is the result of therapeutic resistance and infiltrating growth into the surrounding brain. Here we present a protocol for the detection of the cytoskeleton intermediate filament, vimentin, in cells at the proliferating spheroid surface. By combining a classical invasion assay with immunofluorescence and light-sheet imaging, we find that it is exactly these cytoskeleton-reinforcing cells on the spheroid's surface that will start the infiltration. We anticipate our results to be the starting point of more sophisticated investigation of anti-cancer drug effects on cytoskeleton reorganisation.