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Related Concept Videos

Immunofluorescence Microscopy01:12

Immunofluorescence Microscopy

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A fluorescence microscope uses fluorescent chromophores called fluorochromes, which can absorb energy from a light source and then emit this energy as visible light. Fluorochromes include naturally fluorescent substances (such as chlorophylls) and fluorescent stains that are added to the specimen to create contrast. Dyes such as Texas red and FITC are examples of fluorochromes. Other examples include the nucleic acid dyes 4’,6’-diamidino-2-phenylindole (DAPI), and acridine orange.
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Phagocytosis00:41

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Cells pull particles inward and engulf them in spherical vesicles in an energy-requiring process called endocytosis. Phagocytosis (“cellular eating”) is one of three major types of endocytosis. Cells use phagocytosis to take in large objects—such as other cells (or their debris), bacteria, and even viruses.
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High Throughput Fluorometric Technique for Assessment of Macrophage Phagocytosis and Actin Polymerization
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Quantifying Phagocytosis by Immunofluorescence and Microscopy.

Christopher H Choy1,2, Roberto J Botelho3,4

  • 1Molecular Science Graduate Program, Ryerson University, Toronto, ON, Canada, M5B2K3.

Methods in Molecular Biology (Clifton, N.J.)
|November 6, 2016
PubMed
Summary
This summary is machine-generated.

This study introduces a new fluorescence microscopy method to quantify phagocytosis, the process by which immune cells engulf particles. The technique measures how effectively macrophage-like cells ingest antibody-coated beads, aiding immune cell research.

Keywords:
ImmunofluorescenceMicroscopyPhagocytic indexPhagocytosisPhagosomeQuantitation

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Area of Science:

  • Immunology
  • Cell Biology
  • Microscopy

Background:

  • Phagocytosis is a critical cellular process for immune defense, involving actin-driven particle internalization by immune cells.
  • Receptor activation initiates signaling cascades that remodel actin and plasma membranes, influencing phagocytosis efficiency.
  • Understanding these molecular mechanisms is key to modulating phagocytic capacity.

Purpose of the Study:

  • To develop and present a fluorescence microscopy-based technique for quantifying phagocytosis.
  • To establish a reliable method for measuring the rates and capacity of phagocytic processes.
  • To provide a tool applicable to various phagocytes and particles.

Main Methods:

  • Utilized a macrophage-like cell line for phagocytosis experiments.
  • Employed fluorescence microscopy to visualize and quantify particle uptake.
  • Quantified phagocytosis of antibody-opsonized polystyrene beads as a model system.

Main Results:

  • Successfully demonstrated a fluorescence microscopy technique to quantify phagocytosis.
  • The method effectively measured the uptake of antibody-opsonized beads by macrophages.
  • The technique's versatility allows for application to diverse phagocytic scenarios.

Conclusions:

  • The presented fluorescence microscopy method offers a robust way to quantify phagocytosis.
  • This technique can be adapted for studying various phagocytic cells and particles.
  • It serves as a valuable tool for advancing research in cellular immunity and particle uptake mechanisms.