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Flow Cytometric Analysis of Bimolecular Fluorescence Complementation: A High Throughput Quantitative Method to Study Protein-protein Interaction
Published on: August 15, 2013
Two-color, double-slit in vivo flow cytometer.
1Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA. jpnovak@alum.mit.edu
Optics Letters
|October 17, 2007
Summary
A novel two-color in vivo flow cytometer was developed to overcome limitations of single-wavelength systems. This advanced instrument enables real-time, multi-parameter analysis of circulating cells and blood flow in live animals.
Area of Science:
- Biomedical Engineering
- Cell Biology
- Physiology
Background:
- In vivo flow cytometry allows real-time analysis of circulating cells in live animals without invasive procedures.
- Existing single-wavelength in vivo flow cytometers have limitations in detecting and quantifying multiple cell types or parameters simultaneously.
- Understanding cell circulation kinetics and flow dynamics is crucial for various biological and medical investigations.
Purpose of the Study:
- To design and construct a second-generation in vivo flow cytometer with enhanced capabilities.
- To overcome the performance limitations of single-excitation slit, single-wavelength in vivo flow cytometers.
- To enable simultaneous detection and quantification of multiple cell populations and parameters in vivo.
Main Methods:
- Development of a novel in vivo flow cytometer equipped with two laser wavelengths (473 nm and 633 nm) and one or two excitation slits.
- Utilizing fluorescently labeled cells and/or fluorescent protein expression for detection.
- Application of the cytometer for real-time monitoring of cell circulation kinetics and blood flow velocity in live animal models.
Main Results:
- The two-color in vivo flow cytometer successfully acquired circulation kinetics data for distinct cell populations labeled with different markers.
- The system enabled analysis of cell populations labeled with multiple markers and those expressing green-fluorescent protein.
- Accurate determination of arterial red blood cell velocities in mice was achieved using the developed cytometer.
Conclusions:
- The developed two-color in vivo flow cytometer significantly enhances the capabilities for studying cellular dynamics and fluid mechanics in vivo.
- This advanced tool provides a more comprehensive understanding of cell circulation and kinetics compared to previous single-wavelength systems.
- The instrument holds promise for advancing research in areas such as immunology, cancer metastasis, and drug delivery.
