Plug flow cytometry extends analytical capabilities in cell adhesion and receptor pharmacology

Area of Science:

• Biotechnology
• Analytical Chemistry
• Cell Biology

Background:

• Plug flow cytometry is a novel system for automated delivery of precise cell or particle volumes.
• Key features include defined sample volumes and no requirement for pressurized sample vessels.
• This enables direct cell concentration determination and instrument integration.

Purpose of the Study:

• To describe the capabilities of plug flow cytometry for cell analysis.
• To demonstrate its application in cell adhesion assays.
• To showcase its integration with other analytical instruments for pharmacological studies.

Main Methods:

• Utilized human polymorphonuclear neutrophils (PMNs) and Chinese hamster ovary (CHO) cells for adhesion assays.
• Employed fluorescent probes (Fura Red, GFP, indo-1) for cell tracking and calcium determination.
• Integrated plug flow cytometry with a cone-plate viscometer and the High Throughput Pharmacology System (HTPS).

Main Results:

• Directly quantified nonadherent PMNs and adherent PMNs in aggregates.
• Enabled real-time flow cytometric analysis of cell-cell adhesion under uniform shear.
• Rapidly characterized intracellular calcium dose-response profiles for pharmacological studies.

Conclusions:

• Plug flow cytometry enhances analytical capabilities by coupling flow cytometers to other fluidics-based instruments.
• Expanded applications in cell adhesion analysis.
• Facilitated pharmacological characterization of receptor-ligand interactions.