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Related Concept Videos

Flow Cytometry01:23

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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Imaging Flow Cytometry of Multi-Nuclearity.

Ivan A Vorobjev1,2,3,4, Sultan Bekbayev5, Adil Temirgaliyev5

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Multi-nuclearity in cancer cells is linked to poor prognosis. Imaging flow cytometry (IFC) offers a more efficient and less biased method for quantifying multi-nucleated cells compared to traditional microscopy.

Keywords:
Cytochalasin DMicrotubule inhibitorsMitosisMulti-nuclearity

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Area of Science:

  • Cell Biology
  • Cancer Research
  • Toxicology

Background:

  • Multi-nuclearity is a common cellular characteristic in various cancers and is frequently observed during drug toxicity assessments.
  • The formation of multi-nucleated cells arises from disruptions in cell division or cytokinesis, and their prevalence often indicates a poorer prognosis in cancer patients.
  • Traditional microscopy methods for analyzing multi-nuclearity are labor-intensive and prone to user bias, while automated microscopy has limitations in visualizing nuclei in attached cells.

Purpose of the Study:

  • To present a detailed experimental protocol for preparing multi-nucleated cell samples from attached cultures.
  • To introduce an algorithm for the quantitative analysis of multi-nucleated cells using imaging flow cytometry (IFC).
  • To compare the efficacy and limitations of IFC with traditional microscopy techniques for multi-nuclear cell analysis.

Main Methods:

  • Development of a sample preparation protocol for multi-nucleated cells derived from attached cultures.
  • Utilization of imaging flow cytometry (IFC) for high-resolution image acquisition of multi-nucleated cells.
  • Implementation and testing of two distinct algorithms for discriminating between single-nucleus and multi-nucleated cells within the IFC data.

Main Results:

  • IFC enables high-resolution imaging of multi-nucleated cells induced by mitotic arrest (e.g., taxol) or cytokinesis blockade (e.g., cytochalasin D).
  • Two algorithms were developed and applied for the accurate discrimination of single-nucleus and multi-nucleated cells.
  • The study discusses the comparative advantages and disadvantages of IFC analysis versus conventional microscopy for multi-nuclear cell quantification.

Conclusions:

  • Imaging flow cytometry (IFC) provides a powerful and efficient alternative to microscopy for the quantitative analysis of multi-nucleated cells.
  • The described IFC protocol and algorithms facilitate accurate assessment of multi-nuclearity, relevant for cancer research and drug toxicity studies.
  • IFC overcomes limitations of microscopy, offering reduced bias and improved data collection for multi-nucleated cell analysis.