Label‐Free Live‐Cell Imaging of Nucleic Acids Using Stimulated Raman Scattering Microscopy
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Imaging of nucleic acids is important for studying cellular processes such as cell division and apoptosis. A noninvasive label‐free technique is attractive. Raman spectroscopy provides rich chemical information based on specific vibrational peaks. However, the signal from spontaneous Raman scattering is weak and long integration times are required, which drastically limits the imaging speed when used for microscopy. Coherent Raman scattering techniques, comprising coherent anti‐Stokes Raman scattering (CARS) and stimulated Raman scattering (SRS) microscopy, overcome this problem by enhancing the signal level by up to five orders of magnitude. CARS microscopy suffers from a nonresonant background signal, which distorts Raman spectra and limits sensitivity. This makes CARS imaging of weak transitions in spectrally congested regions challenging. This is especially the case in the fingerprint region, where nucleic acids show characteristic peaks. The recently developed SRS microscopy is free from these limitations; excitation spectra are identical to those of spontaneous Raman and sensitivity is close to shot‐noise limited. Herein we demonstrate the use of SRS imaging in the fingerprint region to map the distribution of nucleic acids in addition to proteins and lipids in single salivary gland cells of Drosophila larvae, and in single mammalian cells. This allows the imaging of DNA condensation associated with cell division and opens up possibilities of imaging such processes in vivo.
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Cell organelles were determined based on specific Raman peaks: PBS buffer at 1,637 cm−1; cytoplasm at 1,005, 1,250, 1,659, and 1,445 cm−1; TAGs/PLPs at 1,092, 1,308, 1,445, and 1,661 cm−1; nucleus/DNA at 790 and 1,103 cm−1; cholesterol at 1,069, 1,134, 1,300, and 1,443 cm−1.
“Cell organelles were determined based on the following peaks: PBS buffer - 1,637cm−1 (water peak); cytoplasm - 1,005cm−1 (phenylalanine), 1,250cm−1 (Amide III), 1,659cm−1 (Amide I) and 1,445cm−1 (CH deformations of proteins and lipids);”
From Hyperspectral unmixing for Raman spectroscopy via physics-constrained autoencoders (2024) · cites this paper as [109]
Section 2.2 (page unavailable) assigns peaks near the claimed phenylalanine, Amide I, and DNA values; Section 2.4 uses the phenylalanine peak to image protein distribution. The cited paper does not establish the claimed PBS, TAG/PLP, or cholesterol peak lists, or the 1,250 and 1,445 cm−1 cytoplasm assignments. Its reported peak positions also differ slightly from those quoted. (LLM, conf=0.88)
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- Authors
- Zhang, X., Roeffaers, M. B. J., Basu, S., Daniele, J. R., Fu, D., Freudiger, C. W., Holtom, G. R., & Xie, X. S.
- Affiliations
- Harvard University (US)
- Corresponding author
- Xie, X. Sunney
- Venue
- ChemPhysChem
- Publisher
- Wiley
- Published
- 2012-02-24
- Volume
- 13
- Issue
- 4
- Pages
- 1054-1059
- ISSN
- 1439-4235 (ISSN-L)
- 1439-7641
- PMC
- PMC3516876
- OpenAlex
- W2009345619
- Cited by
- 171
- References
- 30
- Citations by year
- 2026: 10 · 2025: 9 · 2024: 8 · 2023: 4 · 2022: 11 · 2021: 10 · 2020: 9 · 2019: 16 · 2018: 12 · 2017: 11 · 2016: 12 · 2015: 18 · 2014: 23 · 2013: 15 · 2012: 3
- FWCI
- 8.62
- Type
- journal article
- Language
- en
- Open access
- green
- europepmc.org
- Locations
- ChemPhysChem (publishedVersion)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- Digital Access to Scholarship at Harvard (DASH) (Harvard University) (submittedVersion, cc-by)
- Funders
- Bill and Melinda Gates Foundation
- Belgian American Educational Foundation
- US-UK Fulbright Commission
- Fonds Wetenschappelijk Onderzoek
- National Institutes of Health
- National Institute of Biomedical Imaging and Bioengineering (R01EB010244)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Spectroscopy Techniques in Biomedical and Chemical Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Advanced Fluorescence Microscopy Techniques
- Physical Sciences › Physics and Astronomy › Atomic and Molecular Physics, and Optics › Spectroscopy and Quantum Chemical Studies
- Keywords
- stimulated Raman scattering microscopy, label-free live-cell imaging, nucleic acids, DNA condensation, cell division, Drosophila salivary gland, mammalian cells, non-resonant background, single-cell imaging
- MeSH terms
- Animals, DNA, Drosophila melanogaster, Humans, Salivary Glands, Spectrum Analysis, Raman, Cell Line, Tumor, HEK293 Cells
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