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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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Flow Cytometric Analysis of Bimolecular Fluorescence Complementation: A High Throughput Quantitative Method to Study Protein-protein Interaction
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Panel Design and Optimization for Full Spectrum Flow Cytometry.

Laura Ferrer-Font1, Sam J Small2,3, Evelyn Hyde3

  • 1Hugh Green Cytometry Centre, Malaghan Institute of Medical Research, Wellington, New Zealand. Laura.Ferrer@bd.com.

Methods in Molecular Biology (Clifton, N.J.)
|March 25, 2024
PubMed
Summary
This summary is machine-generated.

Full spectrum flow cytometry enables complex immune system analysis with large panels. This study offers optimized protocols for panel design, autofluorescence management, and data analysis in flow cytometry.

Keywords:
Assay optimizationAutofluorescenceFull spectrum flow cytometryHigh-dimensional flow cytometryImmunophenotypingLongitudinal studiesPanel design

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Area of Science:

  • Immunology
  • Biotechnology
  • Analytical Chemistry

Background:

  • Advancements in fluorescence flow cytometry and immunology have driven the creation of large, multi-marker panels for single-cell analysis.
  • Full spectrum flow cytometry, which analyzes the entire emission spectrum of fluorophores, is increasingly adopted globally.
  • The complexity of immune system research necessitates optimized protocols for advanced flow cytometry techniques.

Purpose of the Study:

  • To provide optimized, step-by-step protocols for full spectrum flow cytometry.
  • To guide researchers in panel design, panel optimization, and high-dimensional data analysis.
  • To address challenges in characterizing autofluorescence for improved marker resolution in complex samples.

Main Methods:

  • Development of standardized protocols for full spectrum flow cytometry panel design.
  • Implementation of strategies for autofluorescence evaluation and mitigation.
  • Establishment of methods for optimizing panel performance and data analysis.
  • Guidance on conducting longitudinal studies using full spectrum flow cytometry.

Main Results:

  • Optimized protocols enhance panel design and optimization for complex staining panels.
  • Effective autofluorescence characterization improves marker resolution, especially in challenging samples.
  • The provided methods facilitate robust high-dimensional data analysis in flow cytometry.
  • Protocols support the successful execution of longitudinal studies with full spectrum flow cytometry.

Conclusions:

  • Optimized protocols are essential for maximizing the potential of full spectrum flow cytometry.
  • Addressing autofluorescence is critical for accurate single-cell analysis in complex biological systems.
  • This work provides a comprehensive guide for researchers utilizing advanced flow cytometry techniques.
  • The developed protocols support reproducible and high-quality data generation in immunological studies.