Assessment of GFP expression and viability using the tali image-based cytometer

Area of Science:

• Cell Biology
• Biotechnology
• Analytical Chemistry

Background:

• Traditional flow cytometry and fluorescence microscopy offer distinct but incomplete cellular analysis.
• Flow cytometry provides quantitative data but lacks visualization.
• Fluorescence microscopy offers visual data but limited quantitative analysis.

Purpose of the Study:

• To present a method for cell viability and green fluorescent protein (GFP) expression assays using the Tali Image-Based Cytometer.
• To highlight the advantages of the Tali instrument over conventional methods.
• To demonstrate the comparable performance of image-based cytometry to flow cytometry.

Main Methods:

• Utilized the Tali Image-Based Cytometer for simultaneous bright field, green fluorescence, and red fluorescence imaging.
• Performed cell viability assays using the Tali Viability Kit - Dead Cell Red with propidium iodide (PI).
• Analyzed GFP expression in conjunction with cell viability in GFP-transduced cells.

Main Results:

• The Tali Image-Based Cytometer successfully performed cell viability and GFP expression assays.
• Generated histograms for cell size, PI fluorescence intensity, and GFP fluorescence intensity.
• Demonstrated comparable data to traditional flow cytometry for cell viability and protein expression.

Conclusions:

• Image-based cytometry, exemplified by the Tali instrument, bridges the gap between flow cytometry and fluorescence microscopy.
• The Tali cytometer offers a cost-effective, user-friendly alternative with added visual data.
• This method enables simultaneous quantitative analysis and visualization of heterogeneous cell populations.