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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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Improvement of Bacillus subtilis Spore Enumeration and Label Analysis in Flow Cytometry

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An efficient method for enumerating oral spirochetes using flow cytometry.

Rebecca Orth¹, Neil O'Brien-Simpson, Stuart Dashper

¹Cooperative Research Centre for Oral Health Science, Melbourne Dental School, Australia.

Journal of Microbiological Methods

|November 26, 2009

Summary

This summary is machine-generated.

This study introduces a new oral bacterial growth medium for cultivating Treponema denticola. Flow cytometry is identified as the most accurate and rapid method for enumerating these difficult-to-culture spirochetes.

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Area of Science:

Microbiology
Bacteriology

Background:

Spirochetes, including Treponema denticola, are challenging to enumerate due to their thin cell walls and cultivation difficulties.
Accurate quantification of T. denticola is crucial for understanding oral microbial ecology and disease pathogenesis.

Purpose of the Study:

To develop an improved oral bacterial growth medium (OBGM) for T. denticola cultivation.
To compare three enumeration methods: colony-forming unit (CFU) counts, DNA analysis, and flow cytometry.
To identify the most reliable and efficient method for quantifying T. denticola.

Main Methods:

Cultivation of T. denticola using a modified OBGM.
Enumeration via semi-solid agar CFU counts.
Quantification using DNA analysis.
Assessment of cell density using flow cytometry.

Main Results:

The modified OBGM significantly improved T. denticola cultivation compared to previous media.
Semi-solid agar CFU counts underestimated cell density by 50-fold.
DNA analysis and flow cytometry showed strong linear correlations with culture absorbance (R(2) > 0.93).
Flow cytometry provided accurate cell density estimations (approx. 6.7 x 10(8) cells/mL at A650=0.2) and distinguished live/dead bacteria.

Conclusions:

Flow cytometry is a superior method for rapid and reliable enumeration of viable spirochetes.
The developed OBGM enhances T. denticola cultivation, facilitating further research.
Accurate enumeration techniques are vital for studying spirochete populations in oral environments.