Bio-Rad Gene Pulser MXcell™ Electroporation System - Manual (Page 16)

There are many techniques available to examine cells for transfection efficiency and cell
viability. We have briefly summarized three commonly used methods. It is critical to the
success of your experiment to evaluate all cells that were electroporated.
Fluorescence Microscopy for Adherent Cells
Fluorescence microscopy can be used to visualize a fluorescent signal within the cell.
This method is commonly used when expressing a GFP-tagged protein.
Materials
n
PBS
n
Fixation buffer: 2–4% formaldehyde in PBS
n
70% glycerol
Method
1. Remove the medium from the wells of the electroporation plate and wash the cells
once with 00–700 μl of PBS.
2. Add 00 μl of fixation buffer to each well and incubate at room temperature for 10 min.
. Remove the fixative, and perform 2 washes with PBS.
4. Add 70% glycerol, and store the cells at 4°C until they are ready to be analyzed by
fluorescence microscopy using the appropriate filters.
Flow Cytometry
Flow cytometry can be used to measure the number of cells containing a fluorescent
tag, such as a fluorescent siRNA or a GFP-tagged protein.
Materials
n
Fixation buffer: 2–4% formaldehyde in PBS
n
PBS
n
70% glycerol
Method
1. For adherent cells, add 100 μl trypsin per well to detach cells and add medium
(200–00 μl) to inactivate the trypsin. For cells in suspension, skip this step.
2. Transfer the cell suspension to a 1. ml centrifuge tube and pellet the cells (00 RCF).
. Remove the medium and resuspend the cells in 00 μl PBS.
4. Transfer the resuspended cells to a flow cytometer tube for analysis.
Gene Pulser MXcell Electroporation Guide
Electroporation Evaluation Methods
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