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Flow Cytometry01:23

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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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Image-based Flow Cytometry Technique to Evaluate Changes in Granulocyte Function In Vitro
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Cellometer image cytometry as a complementary tool to flow cytometry for verifying gated cell populations.

Dmitry Kuksin1, Christina Arieta Kuksin2, Jean Qiu1

  • 1Department of Technology R&D, Nexcelom Bioscience, Lawrence, MA, 01843, USA.

Analytical Biochemistry
|April 2, 2016
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Summary

Image cytometry offers a cost-effective and user-friendly alternative to traditional flow cytometry for cell analysis. This automated system provides comparable results for cell vitality and viability measurements, with added visual confirmation capabilities.

Keywords:
Calcein AMCellometer VisionFlow cytometryImage cytometryPropidium iodide

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Area of Science:

  • Cell Biology
  • Biotechnology
  • Analytical Chemistry

Background:

  • Flow cytometry is a standard technique for cell population analysis but is expensive and requires specialized expertise.
  • Emerging image cytometry systems offer automated cell analysis using bright-field and fluorescent imaging.
  • Image cytometry presents a potentially more accessible alternative for cell-based assays.

Purpose of the Study:

  • To compare the effectiveness of a novel image cytometry system against traditional flow cytometry for cell vitality and viability assessment.
  • To evaluate the capabilities of image cytometry in analyzing cell populations based on size and fluorescence intensity.
  • To determine if image cytometry can serve as a complementary tool to flow cytometry.

Main Methods:

  • Cells were labeled with calcein-AM (vitality stain) and propidium iodide (viability dye).
  • Cell populations were analyzed using both a novel image cytometry system and a conventional flow cytometer.
  • Gating operations based on cell size and fluorescence intensity were performed on both platforms.

Main Results:

  • Image cytometry demonstrated comparable effectiveness to flow cytometry in measuring cell vitality and viability.
  • The image cytometer successfully captured bright-field and fluorescent images, enabling size and fluorescence intensity measurements.
  • Visual confirmation of cell samples was possible with image cytometry, aiding in the exclusion of cellular debris.

Conclusions:

  • Image cytometry provides a direct and effective method for fluorescent cell-based assays.
  • This technology offers a user-friendly and potentially more affordable alternative to flow cytometry.
  • Image cytometry can function as a complementary tool to flow cytometry, particularly for complex sample analyses.