Interpreting CARS images of tissue within the C–H‐stretching region

Single band coherent anti-Stokes Raman scattering (CARS) microscopy is one of the fastest implementation of nonlinear vibrational imaging allowing for video-rate image acquisition of tissue. This is due to the large Raman signal in the C-H-stretching region. However, the chemical specificity of such images is conventionally assumed to be low. Nonetheless, CARS imaging within the C-H-stretching region enables detection of single cells and nuclei, which allows for histopathologic grading of tissue. Relevant information such as nucleus to cytoplasm ratio, cell density, nucleus size and shape is extracted from CARS images by innovative image processing procedures. In this contribution CARS image contrast within the C-H-stretching region is interpreted by direct comparison with Raman imaging and correlated to the tissue composition justifying the use of CARS imaging in this wavenumber region for biomedical applications.

[1]  Cesar Jauregui,et al.  All-fiber laser source for CARS microscopy based on fiber optical parametric frequency conversion. , 2012, Optics express.

[2]  Bruce J Tromberg,et al.  The need for speed , 2012, Smart Structures.

[3]  Stephen T. C. Wong,et al.  Label-free high-resolution imaging of prostate glands and cavernous nerves using coherent anti-Stokes Raman scattering microscopy , 2011, Biomedical optics express.

[4]  T. Kelf,et al.  Scar tissue classification using nonlinear optical microscopy and discriminant analysis , 2012, Journal of biophotonics.

[5]  Jürgen Popp,et al.  Towards automated segmentation of cells and cell nuclei in nonlinear optical microscopy. , 2012, Journal of biophotonics.

[6]  B. Dietzek,et al.  Raman and CARS microspectroscopy of cells and tissues. , 2009, The Analyst.

[7]  Alejandro Garcia-Uribe,et al.  Quantitative photoacoustic microscopy of optical absorption coefficients from acoustic spectra in the optical diffusive regime. , 2012, Journal of biomedical optics.

[8]  Jürgen Popp,et al.  Unsupervised unmixing of Raman microspectroscopic images for morphochemical analysis of non-dried brain tumor specimens , 2012, Analytical and Bioanalytical Chemistry.

[9]  Raghuveer Parthasarathy,et al.  Comparing phototoxicity during the development of a zebrafish craniofacial bone using confocal and light sheet fluorescence microscopy techniques , 2013, Journal of biophotonics.

[10]  Stephen T. C. Wong,et al.  Differential diagnosis of breast cancer using quantitative, label-free and molecular vibrational imaging , 2011, Biomedical optics express.

[11]  Jürgen Popp,et al.  Multimodal nonlinear microscopic investigations on head and neck squamous cell carcinoma: Toward intraoperative imaging , 2013, Head & neck.

[12]  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 .

[13]  P. Prasad,et al.  Nonlinear optical imaging and Raman microspectrometry of the cell nucleus throughout the cell cycle. , 2010, Biophysical journal.

[14]  Jürgen Popp,et al.  Nonlinear microscopy, infrared, and Raman microspectroscopy for brain tumor analysis. , 2011, Journal of biomedical optics.

[15]  Cesar Jauregui,et al.  Widely tuneable fiber optical parametric amplifier for coherent anti-Stokes Raman scattering microscopy. , 2012, Optics express.

[16]  J. Bouquot,et al.  Oral epithelial dysplasia classification systems: predictive value, utility, weaknesses and scope for improvement. , 2008, Journal of oral pathology & medicine : official publication of the International Association of Oral Pathologists and the American Academy of Oral Pathology.

[17]  Chi-Kuang Sun,et al.  Characterization of oral squamous cell carcinoma based on higher‐harmonic generation microscopy , 2012, Journal of biophotonics.

[18]  S. Wiberley,et al.  METHYL AND METHYLENE GROUPS , 1990 .

[19]  Jürgen Popp,et al.  From molecular structure to tissue architecture: collagen organization probed by SHG microscopy , 2013, Journal of biophotonics.

[20]  Annika Enejder,et al.  Non‐linear microscopy of smooth muscle cells in artificial extracellular matrices made of cellulose , 2012, Journal of biophotonics.

[21]  Jane A Dickerson,et al.  Current Applications of Liquid Chromatography / Mass Spectrometry in Pharmaceutical Discovery After a Decade of Innovation , 2008 .

[22]  S. Wiberley,et al.  Introduction to infrared and Raman spectroscopy , 1965 .

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

[24]  Jürgen Popp,et al.  Accumulating advantages, reducing limitations: Multimodal nonlinear imaging in biomedical sciences – The synergy of multiple contrast mechanisms , 2013, Journal of biophotonics.

[25]  John Paul Pezacki,et al.  Chemical contrast for imaging living systems: molecular vibrations drive CARS microscopy , 2011, Nature chemical biology.

[26]  X. Xie,et al.  Optical heterodyne-detected Raman-induced Kerr effect (OHD-RIKE) microscopy. , 2011, The journal of physical chemistry. B.

[27]  Julian Moger,et al.  Exploring uptake mechanisms of oral nanomedicines using multimodal nonlinear optical microscopy , 2012, Journal of biophotonics.

[28]  F. Cordelières,et al.  A guided tour into subcellular colocalization analysis in light microscopy , 2006, Journal of microscopy.

[29]  Conor L Evans,et al.  Chemical imaging of tissue in vivo with video-rate coherent anti-Stokes Raman scattering microscopy. , 2005, Proceedings of the National Academy of Sciences of the United States of America.