Immunostaining protocol for infiltrating brain cancer spheroids for light-sheet imaging.
Benedicte BjørknesOliver Emil NeyePetra HamerlikLiselotte JauffredPublished in: PloS one (2023)
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.
Keyphrases
- white matter
- poor prognosis
- induced apoptosis
- cell cycle arrest
- long non coding rna
- high resolution
- randomized controlled trial
- multiple sclerosis
- resting state
- endoplasmic reticulum stress
- emergency department
- high throughput
- papillary thyroid
- oxidative stress
- brain injury
- photodynamic therapy
- squamous cell
- drug induced
- label free
- cell proliferation
- lymph node metastasis
- sensitive detection
- childhood cancer