In this experiment, the Celigo™ Image Cytometer was used to image and quantify lentiviral-GFP transduction efficiencies in 96-well plates at different Multiplicity of Infections (MOIs).
- HEK293T cells were seeded in a 96-well plate and incubated overnight to reach 80% confluency
- 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
- The Celigo was used to image and analyze the cells in brightfield and green fluorescence to measure transduction efficiencies at different MOIs.
- 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)
Figure 1. Brightfield and fluorescent images of HEK293T cells transduced by the lentivirus showing greater GFP fluorescence as virus volume increased.
Figure 2. The dose-dependent curve of GFP+ percentages with respect to virus volume indicating the transduction efficiencies of the lentivirus.
For research use only. Not for use in diagnostic procedures.