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

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Citar este artículo
Immunofluorescence Assay: A Method to Identify Tumor Cells Captured on a Medical Wire. J. Vis. Exp. (Pending Publication), e20245, doi: (2023).

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