Quantitative Chemical Imaging with Multiplex Stimulated Raman Scattering Microscopy

Fu, Dan; Lü, Fan; Zhang, Xu; Freudiger, Christian W.; Pernik, Douglas R.; Holtom, Gary R.; Xie, X. Sunney

doi:10.1021/ja210081h

Cited by 266 publications

(232 citation statements)

References 23 publications

Supporting

Mentioning

224

Contrasting

Order By: Relevance

“…Recent developments on multiplex detection allow for distinguishing various chemical species with overlapping Raman bands by either broadband excitation (16,17) or narrowband scanning (18,19). SRS at two specific Raman shifts within the broadband of the carbon-hydrogen (CH) stretching vibrational mode (2,800-3,050 cm −1 ) has been used to simultaneously map protein and lipid distribution in cells and tissues (20,21).…”

mentioning

confidence: 99%

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

Lü

Basu

Igras

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

242

213

View full text Add to dashboard Cite

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.

show abstract

mentioning

confidence: 99%

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

Lü

Basu

Igras

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

242

213

View full text Add to dashboard Cite

show abstract

“…As SRS is background-free and the signal scales linearly with number of molecules sampled, it is suitable for quantitative imaging. Fu et al showed quantitative multiplexed SRS was possible in micro-algal cell cultures where concentrations of protein, lipid and chlorophyll/carotenoid pigments were measured [254]. Lipid storage in C. elegans was also imaged with SRS and fluorescence microscopy allowing fat-regulatory genes to be screened with quantitative lipid-distribution information.…”

Section: Applicationsmentioning

confidence: 99%

Raman spectroscopy: techniques and applications in the life sciences

Shipp¹,

Sinjab²,

Notingher³

2017

Adv. Opt. Photon.

233

189

View full text Add to dashboard Cite

Raman spectroscopy is an increasingly popular technique in many areas including biology and medicine.It is based on Raman scattering, a phenomenon in which incident photons lose or gain energy via interactions with vibrating molecules in a sample. These energy shifts can be used to obtain information regarding molecular composition of the sample with very high accuracy. Applications of Raman spectroscopy in the life sciences have included quantification of biomolecules, hyperspectral molecular imaging of cells and tissue, medical diagnosis, and others. This review briefly presents the physical origin of Raman scattering explaining the key classical and quantum mechanical concepts. Variations of the Raman effect will also be considered, including resonance, coherent, and enhanced Raman scattering. We discuss the molecular origins of prominent bands often found in the Raman spectra of biological samples. Finally, we examine several variations of Raman spectroscopy techniques in practice, looking at their applications, strengths, and challenges. This review is intended to be a starting resource for scientists new to Raman spectroscopy, providing theoretical background and practical examples as the foundation for further study and exploration.

show abstract

“…imaging of C-H stretching vibration present in the lipid bilayer of the cell membranes [101][102][103]. Two other Raman imaging techniques with great potential have evolved recently: Stimulated Raman scattering spectroscopy and Ultrafast Raman loss spectroscopy [104][105][106][107].…”

Section: Resonance Raman Spectroscopy Surface Enhanced Raman Spectromentioning

confidence: 99%