Cellaca® PLX image cytometer as an alternative for immunophenotyping, GFP/RFP transfection efficiencies, and...

Area of Science:

• Biotechnology
• Cell Biology
• Immunology

Background:

• Cell and gene therapy requires precise instrumentation for cancer therapeutics.
• Flow cytometry is standard for Chemistry Manufacturing and Controls (CMC) but lacks sensitivity for some assays.
• Image cytometry offers cell characterization but has historically lacked sensitivity for surface marker detection.

Purpose of the Study:

• To develop and validate the Cellaca® PLX image cytometry system for routine cell characterization.
• To assess the system's performance for immunophenotyping, transfection/transduction efficiency, and cell health analysis.
• To compare the Cellaca® PLX system directly against a standard flow cytometer (CytoFLEX).

Main Methods:

• Developed methodologies for the Cellaca® PLX system.
• Performed immunophenotyping on peripheral blood mononuclear cells (PBMCs) using T-cell surface markers.
• Analyzed GFP/RFP expressing cell lines for transfection/transduction efficiency and viability.
• Assessed apoptotic markers (annexin V, caspase-3) in treated Jurkat cells.

Main Results:

• Immunophenotyping results for CD3, CD4, and CD8 populations were highly comparable (within 5%) to flow cytometry.
• Transfection/transduction efficiencies and cell viabilities were equivalent between the image cytometry system and flow cytometry.
• Apoptotic marker analysis showed comparable results (within 5%) for annexin V and caspase-3 positive cells.

Conclusions:

• The Cellaca® PLX image cytometry system demonstrates comparable performance to flow cytometry for key cell characterization assays.
• This system may serve as a valuable complementary tool for cell and gene therapy applications.
• The developed system offers potential for improved efficiency and sensitivity in routine cell-based experiments.