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Flow Cytometry

Flow Cytometry

The development of flow cytometry techniques began in 1934 with initial attempts by Andrew Moldavan, a bacteriologist who counted the cells in a flowing capillary system. Moldavan pumped cells through a capillary tube focused under a microscope for visualization. The invention of photometry allowed the measurement of differentially-stained cells, and Louis Kamentsky developed the first multiparameter flow cytometer in 1965 to identify and count the cancer cells in cervical tissue specimens.
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Related Experiment Video

Updated: May 25, 2025

Molecular Probe Optimization to Determine Cell Mortality in a Photosynthetic Organism Microcystis aeruginosa Using Flow Cytometry

Molecular Probe Optimization to Determine Cell Mortality in a Photosynthetic Organism Microcystis aeruginosa Using Flow Cytometry

Published on: January 29, 2016

Enhancing microalgae cell inactivation through hydrodynamic cavitation: Insights from flow cytometry analysis.

Diyuan Wang¹, Francis L de Los Reyes¹, Joel J Ducoste¹

¹Department of Civil, Construction, and Environmental Engineering, North Carolina State University, 915 Partners Way, Campus Box 7908, Raleigh, NC, 27606, USA.

|February 27, 2025

Summary

Hydrodynamic cavitation effectively disrupts microalgae cells. Maintaining cell debris below 20% is crucial for efficient inactivation, with cell membrane integrity as a key optimization target.

Keywords:

Cavitation Cell viability Chlorophyll fluorescence Flow cytometry Microalgae Reactive oxygen species (ROS)

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Published on: August 14, 2020

Area of Science:

Biotechnology
Bioprocess Engineering
Microalgae Cultivation

Background:

Hydrodynamic cavitation (HC) offers a low-energy alternative for microalgae cell disruption.
HC efficiency varies with microalgae species and operating conditions.
Quantifying cellular damage and optimizing HC parameters are critical for bioproduct harvesting.

Purpose of the Study:

To investigate the time-dependent effects of HC on Dunaliella viridis.
To quantify cell disruption and inactivation kinetics under HC.
To identify key physiological endpoints for optimizing HC treatment.

Main Methods:

A bench-top cavitation system was used for multiple HC passes.
Cell concentration, size distribution, and counts were analyzed via cell counter and flow cytometry.
Cell viability, metabolic activity, and reactive oxygen species (ROS) were assessed using fluorescent probes (FDA, EB, DCFDA).

Main Results:

HC effectively disrupted and inactivated Dunaliella viridis cells, following pseudo-first-order kinetics.
Inactivation rate and energy efficiency decreased with increasing cell debris, necessitating debris levels below 10-20%.
Cell membrane rupture was the primary mechanism of inactivation, releasing intracellular contents.

Conclusions:

Cell membrane integrity is a critical endpoint for optimizing HC protocols in microalgae bioproduct harvesting.
A P-factor model improved prediction of cell inactivation kinetics compared to cavitation number alone.
HC inactivates cells by rupturing membranes, with reduced ROS in viable cells.