This video describes the Circulating Tumor Cells (CTC) Immunofluorescence assay, a technique to characterize the biomarkers on circulating tumor cells in non-small cell lung cancer patient samples. This assay could be used to monitor dynamic changes of the tumor and immune response, which may reflect the response to chemotherapy, radiation, and possible immunotherapy treatments.
Protocol
1. Immunofluorescence Staining Enumerate on a chambered slide with a hemocytometer-type grid the number of cells per mL. Dilute the enriched sample with 0.2% anti-binding solution (Table of Materials) to a concentration reaching 100,000 cells/100 µL per cytospin. Moisten the contour of the sample chamber using cotton (Table of Materials) with 50 µL of 0.2% anti-binding solution. Place a polylysine glass-slide in the sample chamber and close. Coat…
Declarações
The authors have nothing to disclose.
Materials
Pluronic F-68 10%
Gibco
24040-032
Anti-binding solution. Storage conditions: Room temperature
EZ Cytofunnels
ThermoFisher
A78710003
Sample chamber with cotton. Storage conditions: Room temperature
Shandon Cytopsin4 centrifuge
ThermoFisher
A78300003
Dedicated centrifuge. Storage conditions: Room temperature
Paraformaldehyde 16%
ThermoFisher
11490570
Fixation solution. Storage conditions: +4°C
Phosphate Buffered Saline Ultra Pure Grade 1X – 1L