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Microscopy & Imaging JoVE (Open Access) Citable · DOI

Cell Electrofusion Visualized with Fluorescence Microscopy

DOI: 10.3791/1991-v
What you'll learn
  • ✓ Load cells with fluorescent trackers CMFDA and CMRA for visualization
  • ✓ Perform electrofusion of adherent cells in vitro using electroporation
  • ✓ Quantify fusion efficiency by fluorescence microscopy image analysis
Protocol

In this video we demonstrate efficient electrofusion of cells in vitro by means of modified adherence method using electroporation and the subsequent detection of fused cells visualization with fluorescence microscopy.

Difficulty
intermediate
Total time
~1-2 hours per batch of cells

Steps

1
Load cells with fluorescent cell trackers

Incubate cells with CMFDA and CMRA fluorescent dyes to label cell populations with distinct colors. This allows visualization of individual cells and identification of fused cells post-electrofusion.

▶ 00:20
2
Perform electrofusion of labeled cells

Apply electrical pulses to adherent cells using a modified adherence electrofusion method to induce membrane fusion between adjacent cells. Monitor fusion process visually during electroporation.

▶ 02:31
3
Acquire fluorescence images and measure fusion yield

Capture fluorescence microscopy images of fused cells and analyze colocalization of both fluorescent trackers within single cells to quantify fusion efficiency and determine overall fusion yield.

▶ 03:39
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