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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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Flow Cytometric Analysis of Biomarkers for Detecting Human Sperm Functional Defects

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Using flow cytometry to estimate pollen DNA content: improved methodology and applications.

Paul Kron¹, Brian C Husband

¹Department of Integrative Biology, University of Guelph, Guelph, Ontario, Canada. pkron@uoguelph.ca

Annals of Botany

|August 10, 2012

Summary

A new filter bursting method efficiently extracts pollen nuclei for flow cytometry, improving genome size and unreduced gamete estimations across many plant species.

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Area of Science:

Plant biology
Cytometry
Genomics

Background:

Flow cytometry is used to analyze pollen nuclear DNA content for plant development studies.
Previous methods for pollen nuclei extraction face challenges, impacting data quality.

Purpose of the Study:

To introduce a simple, novel method for pollen nuclei extraction using filter bursting.
To compare this method with existing techniques and assess its utility for genome size and cell cycle analysis.

Main Methods:

The filter bursting method was tested on 80 species.
Pollen nuclei extraction was compared to chopping and sonication methods.
Genome size estimates from pollen and leaf nuclei were compared.

Main Results:

The filter bursting method achieved high data quality for genome size estimation (81%) and cell cycle analysis (51%).
It yielded higher quality histograms and nuclei yields compared to chopping and sonication.
Pollen-based genome size estimates closely matched leaf tissue estimates.

Conclusions:

The filter bursting method is easy to use, broadly applicable, and successful.
This technique enhances the use of pollen for accurate genome size estimation.
It significantly improves the estimation of unreduced pollen production using flow cytometry.