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Flow Cytometry Assignment

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3 views

Flow Cytometry Assignment

Uploaded by

Aleeha
Copyright
© © All Rights Reserved
Available Formats
Download as DOCX, PDF, TXT or read online on Scribd
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Flow Cytometry Assignment

1. Principles of Flow Cytometer


Flow cytometry is a laser-based technology used to analyze the physical and chemical
properties of particles or cells in suspension. The fluidic system directs single cells through
the laser beam. The optical system, consisting of lasers and filters, detects scattered light
and fluorescence signals. Signal and pulse processing translates these into digital data.
Electrostatic cell sorting isolates specific populations based on fluorescence characteristics.
(Refer to schematic setup for visual understanding).

2. Principles of Fluorescence
Fluorescence occurs when a fluorophore absorbs light at one wavelength and emits light at
a longer wavelength. Fluorescent dyes, such as FITC and PE, are commonly used as markers
to label specific biomolecules, aiding in the detection of cellular components or functions in
flow cytometry.

3. Data Analysis
In flow cytometry, gates and regions are placed to analyze lysed whole blood cells. For
example, lymphocyte, monocyte, and granulocyte populations can be distinguished using
forward and side scatter plots (Refer Fig. 13). Backgating helps identify subsets, such as
leukocytes (Refer Fig. 18). Contour plots (Refer Fig. 14) allow for multi-parametric data
analysis, while univariate histograms (Refer Fig. 15) visualize single-parameter data.

4. Common Applications and New Technology


Flow cytometry is widely used for immunophenotyping, such as identifying major cell
subsets in peripheral blood (Refer Fig. 45, nine-color staining panel). It is also essential for
analyzing cellular functions like cell sorting, regulated cell death, and proliferation. Recent
advancements include multiplexing and high-throughput capabilities.

5. Recent Progress in Flow Cytometry Innovations


Recent innovations in flow cytometry have focused on developing efficient fluorochromes
like quantum dots (QDs), which offer brighter signals and improved photostability. These
advancements enhance multiplexing capabilities and expand applications in cellular and
molecular research.

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