Direct measurement of lentiviral dose-dependent transduction efficiencies using GFP fluorescence

In this experiment, Celigo was used to image and quantify lentiviral-GFP transduction efficiencies in 96-well plates at different MOIs.

1. HEK293T cells are seeded in each well of a 96-well plate and incubated overnight to reach 80% confluency
2. The cells are 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 plate imager imaged and analyzed in brightfield and green fluorescence to measure transduction efficiencies at different MOIs
4. The software automatically identifies and segments cells to provide accurate 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+ positive 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.

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