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Related Concept Videos

Flow Cytometry01:23

Flow Cytometry

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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Protein Dynamics in Living Cells01:19

Protein Dynamics in Living Cells

Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...

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Fluorescence Lifetime Imaging of Molecular Rotors in Living Cells
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Published on: February 9, 2012

Time-resolved fluorescence-decay measurement and analysis on single cells by flow cytometry.

C Deka, J A Steinkamp

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    A new method analyzes fluorescence decays in single cells and particles using flow cytometry and time-domain fluorescence-lifetime spectroscopy. This technique offers adjustable excitation parameters for diverse fluorescence lifetime measurements.

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    Area of Science:

    • Biophysics
    • Analytical Chemistry
    • Cell Biology

    Background:

    • Fluorescence spectroscopy is crucial for analyzing cellular and particle properties.
    • Existing methods may lack the speed for high-throughput analysis or the precision for detailed decay measurements.
    • Time-domain fluorescence-lifetime spectroscopy (TD-FLS) provides detailed decay information but can be slow.

    Purpose of the Study:

    • To develop a novel method for measuring and analyzing fluorescence decays of individual cells and particles in flow.
    • To integrate the speed of flow cytometry with the analytical power of TD-FLS.
    • To enable adaptable measurements for a wide range of fluorescence lifetimes.

    Main Methods:

    • A continuous wave (cw) laser is utilized for excitation.
    • An electro-optic modulator is employed to pulse modulate the laser.
    • The system combines flow cytometry instrumentation with TD-FLS principles.
    • Excitation pulse characteristics and repetition rates are adjustable.

    Main Results:

    • The described method successfully measures fluorescence decays from individual cells and particles in a flow stream.
    • The system's design allows for flexibility in accommodating various fluorescence lifetimes.
    • The integration of flow cytometry ensures rapid sample processing.

    Conclusions:

    • This novel approach provides a powerful tool for high-throughput analysis of fluorescence lifetimes in biological and material samples.
    • The method enhances the capability to study cellular and particle dynamics through fluorescence decay analysis.
    • The adjustable parameters make the system versatile for diverse research applications in life sciences and material science.