The Celigo™ image cytometer can measure the percentage of GFP+ cells and the mean fluorescent intensity distributions across different dilutions. The ability to rapidly assess serial dilution gives confidence to assays that are inherently variable.
In this experiment, the Celigo was used to image and quantify GFP expression after the target cells were treated with different concentrations of sera.
- The target cells were seeded in a 96-well plate and allowed to adhere overnight
- Cells were transduced with AAV-A vector to mid-Multiplicity of Infection (MOI)
- Cells were pre-incubated overnight with different dilutions of serum (1X, 10X, and 100X)
- The Celigo was used to image and analyze the cells in brightfield and GFP fluorescence to determine transduction efficiencies
Serum samples exerted dose-dependent neutralization effects on transduction efficiencies
The Celigo was able to directly measure the GFP+ percentages in the presence or absence of different concentrations of sera. The GFP fluorescent images and analysis results are shown in Figure 1 and Figure 2 respectively.
- The GFP fluorescent images clearly showed that 1X dilution serum samples dramatically reduced GFP+ percentages (<1%)
- The graph confirms that the percentage of GFP-expressing cells was slightly higher for the 10X dilution (~5% of cells)
- GFP+ percentages in wells treated with 100X serum dilution were similar to that of the positive control (16-18% of cells)
Figure 1. GFP fluorescent images for negative and positive controls, sera and 1X, 10X, and 100X dilutions.
Figure 2. GFP+ percentage of target cells demonstrating close-dependent transduction efficiencies.
For research use only. Not for use in diagnostic procedures.