Immunostaining protocol for infiltrating brain cancer spheroids for light-sheet imaging
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Immunostaining protocol for infiltrating brain cancer spheroids for light-sheet imaging. / Bjorknes, Benedicte; Neye, Oliver Emil; Hamerlik, Petra; Jauffred, Liselotte.
In: PLoS ONE, Vol. 18, No. 2, 0281161, 09.02.2023.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - Immunostaining protocol for infiltrating brain cancer spheroids for light-sheet imaging
AU - Bjorknes, Benedicte
AU - Neye, Oliver Emil
AU - Hamerlik, Petra
AU - Jauffred, Liselotte
PY - 2023/2/9
Y1 - 2023/2/9
N2 - 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.
AB - 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.
KW - TO-MESENCHYMAL TRANSITION
KW - INTERMEDIATE-FILAMENTS
KW - VIMENTIN
KW - INVASION
U2 - 10.1371/journal.pone.0281161
DO - 10.1371/journal.pone.0281161
M3 - Journal article
C2 - 36757917
VL - 18
JO - PLoS ONE
JF - PLoS ONE
SN - 1932-6203
IS - 2
M1 - 0281161
ER -
ID: 338299327