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Microfluidic CARS cytometry.

Han-Wei Wang¹, Ning Bao, Thuc L Le

¹Weldon School of Biomedical Engineering, Purdue University, West Lafayette, IN 47907, USA.

|June 11, 2008

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Coherent anti-stokes Raman scattering (CARS) flow cytometry uses a microfluidic device for quantitative analysis. This vibrational spectroscopy method accurately measures adipocyte size distribution in biological samples.

Area of Science:

Biomedical Engineering
Analytical Chemistry
Spectroscopy

Background:

Flow cytometry is a crucial technique for cell analysis.
Coherent anti-stokes Raman scattering (CARS) microscopy offers vibrational contrast for label-free imaging.
Microfluidic devices enable precise control over cellular samples.

Purpose of the Study:

To develop and demonstrate a novel Coherent anti-stokes Raman scattering (CARS) flow cytometry system.
To integrate CARS microscopy with microfluidic technology for enhanced cellular analysis.
To validate the system's capability for quantitative measurements of biological particles.

Main Methods:

Combined a laser-scanning CARS microscope with a polydimethylsiloxane (PDMS) microfluidic device.

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Employed hydrodynamic focusing for sample stream alignment.

Utilized line-scanning across the focused stream for detection of flowing objects.

Optimized parameters using polystyrene beads as standard flowing particles.

Main Results:

Successfully demonstrated Coherent anti-stokes Raman scattering (CARS) flow cytometry.
Validated the system's ability to detect and quantify flowing microparticles.
Performed population measurements of adipocytes isolated from mouse fat tissue.
Showcased accurate quantitation of adipocyte size distribution.

Conclusions:

Microfluidic CARS flow cytometry is a viable new modality for quantitative analysis.
The technique offers vibrational selectivity for label-free cellular characterization.
This method provides a powerful tool for analyzing biological samples like adipocytes.