Immunofluorescence Assay: A Method to Identify Tumor Cells Captured on a Medical Wire
Published: April 30, 2023
Abstract
Source: Gallerani, G. et.al., Characterization of Tumor Cells Using a Medical Wire for Capturing Circulating Tumor Cells: A 3D Approach Based on Immunofluorescence and DNA FISH. J. Vis. Exp. (2017).
This video describes the technique of immunofluorescence staining of circulating tumor cells captured on a functionalized medical wire. The functionalized wire allows isolation of CTCs in vivo, directly from patient’s blood stream.
Protocol
1. Immuno-DNA FISH on Wire Cell line spike-in on wire (3D support) NOTE: Preliminary set-up involves an accurate selection of adherent cells lines based on EpCAM-positivity. The wire is functionalized with anti-EpCAM antibodies so that only EpCAM-positive cells are stained. NOTE: Do not touch the functionalized part of the wire to avoid any damage. NOTE: All steps should be carried out under a laminar flow hood in sterile…
Divulgaciones
The authors have nothing to disclose.
Materials
| Ethanol | Carlo Erba | # 414605 | |
| Tween 20 | BIO RAD | # 1706531 | |
| PBS (Phosphate Buffered Saline) | Medicago | # 09-9400-100 | |
| Acetone | Sigma Aldrich | # 534064-500ML | |
| RUBBERCEMENT | Royal Talens | # 95306500 | |
| EpCAM-FITC (clone: HEA-125) | Mitenyi | #130-080-301 | |
| RPMI 1640 | Gibco | # 31870-025 | |
| XT ALK BA | MetaSystems Probes | # D-6001-100-OG | |
| Detektor CANCER01 EpCAM | Gilupi Nanomedizin | Functionalized wire | |
| ZEISS Fluorescent Microscope Axioskop | ZEISS | ||
| Nikon NIS-Elements BR 4.11.00 64 bit software | Nikon | ||
| Nikon DS-QiMc 12 bit digital camera | Nikon | DS-QiMc |
Citar este artículo
Immunofluorescence Assay: A Method to Identify Tumor Cells Captured on a Medical Wire. J. Vis. Exp. (Pending Publication), e20245, doi: (2023).