1The Univ. of Tokyo (Japan) 2Research Ctr. for Spectrochemistry, The Univ. of Tokyo (Japan) 3PRESTO, Japan Science and Technology Agency (Japan) 4Algal Bio Co., Ltd. (Japan) 5Univ. of California, Los Angeles (United States) 6Institute of Technological Sciences, Wuhan Univ. (China)
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We present a high-throughput multimodal Raman-fluorescence flow cytometer that produces simultaneous Raman and fluorescence excitation spectra at a demonstrated throughput of 100 events per second. Our method combines Fourier-transform coherent anti-Stokes Raman scattering with Fourier-transform two-photon excitation fluorescence, which produces broadband Raman spectra that span the biological fingerprint region and single-detector multicolor fluorescence via spectral analysis. As a proof-of-principle demonstration, we perform multimodal label-free analysis of stress response in the microalga Haematococcus pluvialis by Raman measurement of its secondary metabolite astaxanthin and two-color fluorescence measurement of chlorophyll and fluorescent chlorophyll catabolites under different stress conditions.
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