scholarly journals Spectrometer-free vibrational imaging by retrieving stimulated Raman signal from highly scattered photons

2015 ◽  
Vol 1 (9) ◽  
pp. e1500738 ◽  
Author(s):  
Chien-Sheng Liao ◽  
Pu Wang ◽  
Ping Wang ◽  
Junjie Li ◽  
Hyeon Jeong Lee ◽  
...  
Keyword(s):  
Raman Scattering ◽  
Stimulated Raman Scattering ◽  
Collection Efficiency ◽  
Mouse Skin ◽  
Spectroscopic Imaging ◽  
Acquisition Time ◽  
Raman Signal ◽  
Stimulated Raman ◽  
Photon Collection

In vivo vibrational spectroscopic imaging is inhibited by relatively slow spectral acquisition on the second scale and low photon collection efficiency for a highly scattering system. Recently developed multiplex coherent anti-Stokes Raman scattering and stimulated Raman scattering techniques have improved the spectral acquisition time down to microsecond scale. These methods using a spectrometer setting are not suitable for turbid systems in which nearly all photons are scattered. We demonstrate vibrational imaging by spatial frequency multiplexing of incident photons and single photodiode detection of a stimulated Raman spectrum within 60 μs. Compared to the spectrometer setting, our method improved the photon collection efficiency by two orders of magnitude for highly scattering specimens. We demonstrated in vivo imaging of vitamin E distribution on mouse skin and in situ imaging of human breast cancerous tissues. The reported work opens new opportunities for spectroscopic imaging in a surgical room and for development of deep-tissue Raman spectroscopy toward molecular level diagnosis.

scholarly journals Label-free DNA imaging in vivo with stimulated Raman scattering microscopy

2015 ◽  
Vol 112 (37) ◽  
pp. 11624-11629 ◽  
Author(s):  
Fa-Ke Lu ◽  
Srinjan Basu ◽  
Vivien Igras ◽  
Mai P. Hoang ◽  
Minbiao Ji ◽  
...  
Keyword(s):  
Raman Scattering ◽  
Cell Function ◽  
Stimulated Raman Scattering ◽  
Mouse Skin ◽  
Imaging Method ◽  
Label Free ◽  
Free Dna ◽  
Dna Imaging ◽  
Stimulated Raman

Label-free DNA imaging is highly desirable in biology and medicine to perform live imaging without affecting cell function and to obtain instant histological tissue examination during surgical procedures. Here we show a label-free DNA imaging method with stimulated Raman scattering (SRS) microscopy for visualization of the cell nuclei in live animals and intact fresh human tissues with subcellular resolution. Relying on the distinct Raman spectral features of the carbon-hydrogen bonds in DNA, the distribution of DNA is retrieved from the strong background of proteins and lipids by linear decomposition of SRS images at three optimally selected Raman shifts. Based on changes on DNA condensation in the nucleus, we were able to capture chromosome dynamics during cell division both in vitro and in vivo. We tracked mouse skin cell proliferation, induced by drug treatment, through in vivo counting of the mitotic rate. Furthermore, we demonstrated a label-free histology method for human skin cancer diagnosis that provides comparable results to other conventional tissue staining methods such as H&E. Our approach exhibits higher sensitivity than SRS imaging of DNA in the fingerprint spectral region. Compared with spontaneous Raman imaging of DNA, our approach is three orders of magnitude faster, allowing both chromatin dynamic studies and label-free optical histology in real time.

In Vivo and in Situ Spectroscopic Imaging by a Handheld Stimulated Raman Scattering Microscope

ACS Photonics ◽  
2017 ◽  
Vol 5 (3) ◽  
pp. 947-954 ◽  
Author(s):  
Chien-Sheng Liao ◽  
Pu Wang ◽  
Chih Yu Huang ◽  
Peng Lin ◽  
Gregory Eakins ◽  
...  
Keyword(s):  
Raman Scattering ◽  
Stimulated Raman Scattering ◽  
Spectroscopic Imaging ◽  
Stimulated Raman

scholarly journals Microsecond scale vibrational spectroscopic imaging by multiplex stimulated Raman scattering microscopy

2015 ◽  
Vol 4 (3) ◽  
pp. e265-e265 ◽  
Author(s):  
Chien-Sheng Liao ◽  
Mikhail N Slipchenko ◽  
Ping Wang ◽  
Junjie Li ◽  
Seung-Young Lee ◽  
...  
Keyword(s):  
Raman Scattering ◽  
Stimulated Raman Scattering ◽  
Spectroscopic Imaging ◽  
Stimulated Raman

Virtual brain tumor histopathology and immunohistochemistry with stimulated raman scattering microscopy.

2017 ◽  
Vol 35 (15_suppl) ◽  
pp. e13511-e13511
Author(s):  
Spencer B Lewis ◽  
Balaji Pandian ◽  
Todd Hollon ◽  
Yashar Niknafs ◽  
Mia Garrard ◽  
...  
Keyword(s):  
Brain Tumor ◽  
Raman Scattering ◽  
Stimulated Raman Scattering ◽  
Frozen Section ◽  
Ease Of Use ◽  
Laser Source ◽  
Raman Signal ◽  
Wild Type ◽  
Spectral Difference ◽  
Stimulated Raman

e13511 Background: Stimulated Raman scattering (SRS) microscopy is a label-free optical technique which uses the chemical composition of fresh tissue to generate image contrast. Prior work has demonstrated exceptional correlation between SRS images and H&E microscopy for detecting brain tumor infiltration ex vivo. However, SRS has not previously been suitable for a clinical environment. Further, SRS has utilized structural, rather than chemical, diagnostic features. We present the first clinical validation of SRS, as well as our work towards mutation-targeted SRS imaging. Methods: Implementation of SRS with a fiber laser source allowed sufficient stability for clinical deployment. Unprocessed specimens from 200 neurosurgical cases were imaged at the CH2 and CH3 vibrational modes, then processed to mimic the appearance of H&E staining. 30 cases were trialed in a simulated intraoperative consultation to assess concordance between SRS and cryosection microscopy. A multi-layer perceptron (MLP) model was then used to predict brain tumor diagnosis based on quantified image attributes. To explore the potential for chemically specific SRS, isogenic lines of normal human astrocyte were prepared, differing by the presence or absence of the R132H mutation in the isocitrate dehydrogenase 1 (IDH1) gene. IDH1 R132H cells accumulate high concentrations of the oncometabolite 2-hydroxyglutarate and display predictably altered metabolism. 200 Raman spectra were collected from 50 wild type (WT) and 50 R132H cells with the aim of detecting these changes. Each cell line was then submitted for targeted metabolomic analysis. Results: Excellent concordance was observed between SRS and frozen section histology ( κ >0.89). The MLP diagnosed brain tumor subtype with 90% accuracy. A robust spectral difference was observed between wild type and R132H cells, which showed concordance with metabolomic data. Conclusions: The diagnostic capability, ease of use and speed of SRS make it well-suited for integration into the operative workflow. Further, because SRS amplifies the Raman signal with no resonant background, the spectral differences observed may allow rapid differentiation of IDH1 WT and R132H cells with SRS.

scholarly journals Video-Rate Molecular Imaging in Vivo with Stimulated Raman Scattering

Science ◽  
2010 ◽  
Vol 330 (6009) ◽  
pp. 1368-1370 ◽  
Author(s):  
B. G. Saar ◽  
C. W. Freudiger ◽  
J. Reichman ◽  
C. M. Stanley ◽  
G. R. Holtom ◽  
...  
Keyword(s):  
Molecular Imaging ◽  
Raman Scattering ◽  
Stimulated Raman Scattering ◽  
Stimulated Raman

scholarly journals Narrow-band random Raman lasing from Rhodamine 6G assisted by cascaded stimulated Raman scattering effect

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Mandana Sadat Hosseini ◽  
Elnaz Yazdani ◽  
Batool Sajad
Keyword(s):  
Raman Scattering ◽  
Rhodamine 6G ◽  
Stimulated Raman Scattering ◽  
Gain Medium ◽  
Single Frequency ◽  
Raman Signal ◽  
Raman Gain ◽  
Disordered Medium ◽  
Srs Generation ◽  
Stimulated Raman

AbstractThis study reports the first experimental observation of cascaded stimulated Raman scattering (SRS) generation in a colloidal disordered medium. Generation of the cascaded effect requires both a high Raman gain and pump power in the disordered medium. Here, to extend effective path lengths of photons into the Raman gain medium for producing additional SRS processes, ZnO microspheres with abundant nano-protrusions as suitable scattering centers are proposed. It is explained that nano-protrusions on the surface of the spheres can act as nano reflectors and significantly provide potent feedback in the disordered system. This provided feedback via nano-protrusions boosts cascaded SRS generation to allow the appearance of higher Raman signals of Rhodamine 6G dye solution at a low scatterer concentration of 5 mg/ml. The threshold for the formation of the first Raman signal is measured at about 60 mJ/pulse. Also, the evolution of Raman signals under several fixed pump pulses is examined to investigate the stability from pulse to pulse. Our findings provide promising perspectives for achieving the single-frequency laser sources and generate desirable wavelengths for specific applications.

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