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Near infrared in vivo flow cytometry for tracking fluorescent circulating cells
Yuanzhen Suo1, Tao Liu2, Chengying Xie1
1Med-X Research Institute and School of Biomedical Engineering, Shanghai Jiao Tong University, Shanghai, 200030, China.
Insights
Near-infrared (NIR) in vivo flow cytometry (IVFC) extends a powerful cell tracking technique. This study demonstrates NIR dye IR-780 for detecting circulating cells and blood flow, enabling multicolor analysis in biomedical research.
Area of Science:
- Biomedical Engineering
- Optical Imaging
- Cell Biology
Background:
- In vivo flow cytometry (IVFC) is crucial for tracking cells in the circulatory system.
- Current fluorescence-based IVFC is limited to the visible spectrum.
- Near-infrared (NIR) dyes offer advantages like deeper tissue penetration and reduced scattering for fluorescence detection.
Purpose of the Study:
- To demonstrate the feasibility of using NIR dyes with IVFC.
- To quantify circulating cells labeled with NIR dye IR-780.
- To extend IVFC capabilities into the NIR spectrum for advanced biomedical research.
Main Methods:
- Utilized an NIR in vivo flow cytometer with 785 nm laser excitation.
- Measured IR-780 labeled circulating cells in vivo.
- Obtained blood flow information, including velocity and volume per unit time.
- Performed simultaneous detection of IR-780 and enhanced green fluorescent protein (EGFP) signals for multicolor analysis.
Main Results:
- Successfully detected and quantified representative peaks of NIR fluorescent circulating cells.
- Acquired blood flow velocity and flow volume data.
- Demonstrated IR-780 as a suitable dye for multicolor IVFC, including NIR detection.
- Confirmed the extension of IVFC into the NIR range.
Conclusions:
- NIR IVFC using IR-780 is a viable technique for in vivo cell tracking and blood flow analysis.
- This advancement expands the capabilities of IVFC for multicolor detection in the NIR spectrum.
- NIR IVFC shows significant potential for applications in biomedical research and diagnostics.
Abstract:
The in vivo flow cytometry (IVFC) is now a powerful technique in biomedical research, especially for tracking specific cells in circulatory system. The current fluorescence-based IVFC is limited to visible spectrum, while near infrared (NIR) dyes have their advantages, such as deeper penetration, less absorption and less scattering for NIR fluorescence. Here, using an NIR in vivo flow cytometer with a 785 nm laser excitation, the measurement of fluorescent dye IR-780 labeled circulating cells is demonstrated. Representative peaks corresponding to NIR fluorescent circulating cells are detected and quantified. In addition, blood flow information, including the blood flow velocity and flow volume per unit time, is obtained. By simultaneous detection of IR-780 and enhanced green fluorescent protein (EGFP) signals from dual labeled cells, the IR-780 is shown to be a suitable fluorescent dye for multicolor detection by IVFC, including NIR. Thus, the IVFC is extended to the NIR range and shows potential application in biomedical research.
