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Overview Of Cell Separation And Isolation

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Cell separation was first achieved in 1964 by S. H. Seal, who separated large tumor cells from the smaller blood cells using filtration. Two years later, Pohl and Hawk performed experiments on how cells respond differently to a nonuniform electric field based on the cell type. Such observations were the inception of cell separation methods, which allow isolating a single cell type from a heterogeneous sample.

Flow Cytometry

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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: Jul 2, 2025

A Combinatorial Single-cell Approach to Characterize the Molecular and Immunophenotypic Heterogeneity of Human Stem and Progenitor Populations

A Combinatorial Single-cell Approach to Characterize the Molecular and Immunophenotypic Heterogeneity of Human Stem and Progenitor Populations

Published on: October 25, 2018

Multiomics Characterization of a Less Invasive Microfluidic-Based Cell Sorting Technique.

Feroza K Choudhury¹, Viji Premkumar¹, Jana Zecha¹

¹Dynamic Omics, Centre for Genomics Research (CGR), Discovery Sciences, BioPharmaceuticals R&D, AstraZeneca, Gaithersburg, Maryland 20878, United States.

Journal of Proteome Research

|February 28, 2024

Summary

Microfluidic chip-based cell sorting minimally impacts cell metabolism and phosphoproteomics compared to traditional droplet-based methods. This advanced technique preserves cellular health and function, offering a less disruptive alternative for cell isolation.

Keywords:

LC-MS Sorting fluorescence-activated cell sorting (FACS)metabolomics multiomics phosphoproteomics sorted cells sorter-induced cellular stress (SICS)stable isotope tracing

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Area of Science:

Cell Biology
Biotechnology
Metabolomics

Background:

Fluorescence-activated cell sorting (FACS) is crucial for isolating cell subpopulations.
Cell sorting procedures can negatively affect cell viability and metabolic state.
Traditional droplet-based cell sorters may induce significant cellular stress.

Purpose of the Study:

To compare the impact of droplet-based vs. microfluidic chip-based cell sorting on cellular metabolism and phosphoproteomics.
To evaluate the recovery of metabolic functions post-cell sorting.
To identify the less disruptive cell sorting technology.

Main Methods:

Comparative analysis of cell sorting techniques: droplet-based vs. microfluidic chip-based.
Assessment of metabolic profiles (redox, energy status) and phosphoproteomics.
¹³C-isotope tracing to analyze metabolic pathway recovery (TCA cycle, biosynthesis).

Main Results:

Microfluidic chip-based sorting preserved metabolic and phosphoproteomic profiles closer to unsorted controls.
Droplet-based sorting induced significant alterations in redox, energy status, and proteostasis.
¹³C-isotope tracing showed faster recovery of TCA cycle and biosynthesis pathways in microfluidic chip-based sorted cells.

Conclusions:

Microfluidic chip-based cell sorting demonstrates minimal impact on cellular metabolism and phosphoproteomics.
Droplet-based cell sorting causes greater metabolic disruption and slower recovery.
Microfluidic chip-based sorting is a less disruptive and preferable method for maintaining cellular integrity post-isolation.