Flow cytometry is a widely used tool for high-throughput quantitative analysis of cell populations and intracellular content. Signals in flow cytometry arise from electrical impedance, forward or side light scattering, and fluorescence. Scattering and electrical impedance provide granularity and size/volume information, but with no chemical specificity. Fluorescent labeling acts as the primary approach for cellular analysis in flow cytometry. Nevertheless, fluorescent tags are not applicable to all cases, especially small molecules (e.g. drugs) for which labeling may significantly perturb their properties. The current application aims to fill this gap through the development of a multichannel spectral flow cytometer using stimulate Raman scattering (SRS) signal from inherent molecular vibration. The stimulated Raman scattering overcomes the low signal level in spontaneous Raman scattering. It measures the light-matter energy transfer and is therefore free of the nonresonant background encountered in coherent anti-Stokes Raman scattering. Moreover, as a nonlinear optical process it is inherently phase matched, permitting a weakly focused collinear beam geometry that is compatible with high-speed detection of flowing objects. The planned instrumentation contains two specific aims. The first is to build a single-frequency SRS flow cytometer using a femtosecond laser source. The second is to build a multichannel SRS flow cytometer by multiplex detection of spectrally dispersed SRS signals. Based on the large signal level, we expect to reach the speed of 10,000 cells per second. Performance of the label-free spectral cytometer will be tested through quantitation of fat storage in adipocytes and of drug uptake by cancer cells.
(provided by applicant): We propose to develop a label-free multichannel spectral flow cytometer using the stimulated Raman scattering signal from inherent molecular vibration. The label-free spectral cytometer will be applied to quantify the fat storage in adipocytes and drug uptake by cancer cells.
|Liao, Chien-Sheng; Slipchenko, Mikhail N; Wang, Ping et al. (2015) Microsecond Scale Vibrational Spectroscopic Imaging by Multiplex Stimulated Raman Scattering Microscopy. Light Sci Appl 4:|
|Liao, Chien-Sheng; Wang, Pu; Wang, Ping et al. (2015) Spectrometer-free vibrational imaging by retrieving stimulated Raman signal from highly scattered photons. Sci Adv 1:e1500738|
|Zhang, Chi; Zhang, Delong; Cheng, Ji-Xin (2015) Coherent Raman Scattering Microscopy in Biology and Medicine. Annu Rev Biomed Eng 17:415-45|
|Hu, Chun-Rui; Zhang, Delong; Slipchenko, Mikhail N et al. (2014) Label-free real-time imaging of myelination in the Xenopus laevis tadpole by in vivo stimulated Raman scattering microscopy. J Biomed Opt 19:086005|
|Zhang, Delong; Wang, Ping; Slipchenko, Mikhail N et al. (2014) Fast vibrational imaging of single cells and tissues by stimulated Raman scattering microscopy. Acc Chem Res 47:2282-90|
|Wang, Ping; Liu, Bin; Zhang, Delong et al. (2014) Imaging lipid metabolism in live Caenorhabditis elegans using fingerprint vibrations. Angew Chem Int Ed Engl 53:11787-92|
|Zhang, Delong; Wang, Ping; Slipchenko, Mikhail N et al. (2013) Quantitative vibrational imaging by hyperspectral stimulated Raman scattering microscopy and multivariate curve resolution analysis. Anal Chem 85:98-106|
|Wang, Ke; Zhang, Delong; Charan, Kriti et al. (2013) Time-lens based hyperspectral stimulated Raman scattering imaging and quantitative spectral analysis. J Biophotonics 6:815-20|
|Wang, Pu; Slipchenko, Mikhail N; Mitchell, James et al. (2013) Far-field Imaging of Non-fluorescent Species with Sub-diffraction Resolution. Nat Photonics 7:449-453|
|Zhang, Delong; Slipchenko, Mikhail N; Leaird, Daniel E et al. (2013) Spectrally modulated stimulated Raman scattering imaging with an angle-to-wavelength pulse shaper. Opt Express 21:13864-74|
Showing the most recent 10 out of 12 publications