Direct Measurement Of Lentiviral Dose-Dependent Transduction Efficiencies Using GFP Fluorescence

In this experiment, the Celigo™ Image Cytometer Celigo Image Cytometer  was used to image and quantify lentiviral-GFP transduction efficiencies in 96-well plates at different Multiplicity of Infections (MOIs).   

1. HEK293T cells were seeded in a 96-well plate and incubated overnight to reach 80% confluency
2. The cells were then transduced with five different viral amounts (1, 0.1, 0.01, 0.001, 0.0001 mL) and incubated to express GFP
3. The Celigo was used to image and analyze the cells in brightfield and green fluorescence to measure transduction efficiencies at different MOIs.
4. The software automatically identified and segmented cells to provide counts of both total cells and GFP+ cells
   Dose-dependent measurement GFP+ percentages indicating lentiviral transduction efficiencies
    

The results showed a dose-dependent response in respect to GFP+ percentages shown in the brightfield and fluorescent images in Figure 1.

• GFP+ cells increased as the volume of viral particles increased (Figure 1)
• AAV-B showed higher gene expression over time in comparison to AAV-D (Figure 2)