Flow cytometry

Introduction
  • Flow cytometry depends on fluid dynamics, electronics and optics in its principle.
  • Its name consists of flow which refers to the motion or fluids, cyto is a synonym of a cell and metry is the process of the measurement, so flow cytometry refers to the measurement of the properties of the cells in the fluid flux.
Photos
Applications

Flow cytometry is a technique used in cell biology, cancer testing and research, stem cell research, including:

  • Cell counting
  • Cell sorting
  • Determining cell characteristics and function
  • Detecting microorganisms
  • Biomarker detection
  • Protein engineering detection
  • Diagnosis of health disorders such as blood cancers
Mechanism of action

As mentioned before there are three principles that the flow cytometry can be explained through:

Fluid Dynamics:

Firstly, a sample is taken to be injected into the sheath fluid gushing under
somewhat high pressure in a laminar flow manner in the flow cytometry causing
the present cells to align in a single row to be drifted into a laser beam.

  • Sheath fluid is the saline buffer.
  • Laminar flow describes the movement of each layer in the flow cytometry
    where all layers move adjacently with almost no mixing.

Optics:

The streaming cells pass through the laser beam individually causing the refraction
of the beam to the detectors that determine the structure of the cell accordingly
which is known as the forward scattering (FSC).

Usually, there is a combination of forward and side scattering (SSC) in order to
determine not only the volume and size of the cells but also its granularity and
nuclear structure.

Electronics:

After the refraction of the laser beam, then comes the role of the detectors to
interpret the signals into understandable data that can be used for the
identification and determination of the exact cell types passing through the sheath
fluid.

Materials and reagents
  • Small Organic Molecules
  • Phycobiliproteins
  • Quantum Dots
  • Polymer Dyes
  • Tandem Dyes
  • Metal Conjugates for Mass Cytometry
  • Fluorescent Proteins
  • Nucleic Acid Dyes
  • Proliferation Dyes
  • Viability Dyes
  • Calcium Indicator Dyes
Troubleshooting
  • Focus on the subtle changes in the intensity of the fluorescence as they can interpret different results.
  • Use a dim fluoro-chrome to reduce errors in channeling.
Results and Calculations

Data analysis in flow cytometry depends on:

  • Compensation
  • Gating
  • Computational analysis
  • Fluorescence minus one (FMO) control

Cell sorting

For specific inquiries
If you are in need of a specific question, you can discuss your inquiry with one of our experts in this field:
Dr. Albert Einstein
Theoretical physicist, widely acknowledged

Phone: 32423742

Address: 5 test st.

Summary
  • Flow cytometry is a technique use to analyze multiple cells rapidly, saving both time and efforts.
  • It can differentiate between the cells through their optical properties.
Further readings

If you want to read more about related topics, you can check:

  • Mass spectroscopy
  • Fluorescence spectrometer
  • Optics
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