Time‐lens based hyperspectral stimulated Raman scattering imaging and quantitative spectral analysis

We demonstrate a hyperspectral stimulated Raman scattering (SRS) microscope through spectral-transformed excitation. The 1064 nm Stokes pulse was from a synchronized time-lens source, generated through time-domain phase modulation of a continuous wave (CW) laser. The tunable pump pulse was from linear spectral filtering of a femtosecond laser output with an intra-pulse spectral scanning pulse shaper. By electronically modulating the time-lens source at 2.29 MHz, hyperspectral stimulated Raman loss (SRL) images were obtained on a laser-scanning microscope. Using this microscope, DMSO in aqueous solution with a concentration down to 28 mM could be detected at 2 μs time constant. Hyperspectral SRL images of prostate cancer cells were obtained. Multivariate curve resolution analysis was further applied to decompose the SRL images into concentration maps of CH₂ and CH₃ bonds. This method offers exciting potential in label-free imaging of live cells using fingerprint Raman bands.

[1]  X. Xie,et al.  Multicolored Stain-free Histopathology with Coherent Raman Imaging , 2012, Laboratory Investigation.

[2]  T. B. Bakker Schut,et al.  Discriminating basal cell carcinoma from perilesional skin using high wave-number Raman spectroscopy. , 2007, Journal of biomedical optics.

[3]  Kazuyoshi Itoh,et al.  Stimulated Raman hyperspectral imaging based on spectral filtering of broadband fiber laser pulses. , 2012, Optics letters.

[4]  Ji‐Xin Cheng,et al.  Heterodyne detected nonlinear optical imaging in a lock‐in free manner , 2012, Journal of biophotonics.

[5]  Kazuyoshi Itoh,et al.  Sensitivity enhancement of fiber-laser-based stimulated Raman scattering microscopy by collinear balanced detection technique. , 2012, Optics express.

[6]  Mortazavi,et al.  Supporting Online Material Materials and Methods Figs. S1 to S13 Tables S1 to S3 References Label-free Biomedical Imaging with High Sensitivity by Stimulated Raman Scattering Microscopy , 2022 .

[7]  W. R. Wiley,et al.  Three-Dimensional Vibrational Imaging by Coherent Anti-Stokes Raman Scattering , 1999 .

[8]  Ping Wang,et al.  Quantitative vibrational imaging by hyperspectral stimulated Raman scattering microscopy and multivariate curve resolution analysis. , 2013, Analytical chemistry.

[9]  X. Xie,et al.  Video-Rate Molecular Imaging in Vivo with Stimulated Raman Scattering , 2010, Science.

[10]  Alberto Diaspro,et al.  Coherent Raman Scattering Microscopy , 2014 .

[11]  Xiaohui Ni,et al.  Multicolor stimulated Raman scattering microscopy , 2012 .

[12]  Dan Fu,et al.  Quantitative chemical imaging with multiplex stimulated Raman scattering microscopy. , 2012, Journal of the American Chemical Society.

[13]  Dan Fu,et al.  Hyperspectral imaging with stimulated Raman scattering by chirped femtosecond lasers. , 2013, The journal of physical chemistry. B.

[14]  Delong Zhang,et al.  Highly Sensitive Vibrational Imaging by Femtosecond Pulse Stimulated Raman Loss. , 2011, The journal of physical chemistry letters.

[15]  Wei Min,et al.  Highly specific label-free molecular imaging with spectrally tailored excitation stimulated Raman scattering (STE-SRS) microscopy. , 2011, Nature photonics.

[16]  M. Handzic.  5 , 1824, The Banality of Heidegger.

[17]  X. Xie,et al.  Coherent Anti-Stokes Raman Scattering Microscopy: Instrumentation, Theory, and Applications , 2004 .

[18]  Christian W. Freudiger,et al.  Synchronized time-lens source for coherent Raman scattering microscopy , 2010, Optics express.

[19]  D. Begley,et al.  Imaging cortical vasculature with stimulated Raman scattering and two‐photon photothermal lensing microscopy , 2012 .

[20]  Kazuyoshi Itoh,et al.  High-speed molecular spectral imaging of tissue with stimulated Raman scattering , 2012, Nature Photonics.

[21]  Srinjan Basu,et al.  Label-free live-cell imaging of nucleic acids using stimulated Raman scattering microscopy. , 2012, Chemphyschem : a European journal of chemical physics and physical chemistry.

[22]  Lingjie Kong,et al.  Multicolor stimulated Raman scattering microscopy with a rapidly tunable optical parametric oscillator. , 2013, Optics letters.

[23]  Chris Xu,et al.  Fiber-delivered picosecond source for coherent Raman scattering imaging. , 2011, Optics letters.