Label-free DNA imaging in vivo with stimulated Raman scattering microscopy
Abstract
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. Moreover, 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. In conclusion, 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 bothmore »
- Authors:
-
- Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA 02138,
- Department of Molecular and Cellular Biology, Harvard University, Cambridge, MA 02138,
- Cutaneous Biology Research Center, Department of Dermatology, Massachusetts General Hospital, Harvard Medical School, Charlestown, MA 02129,
- Department of Pathology, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02114,
- Department of Dermatology, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02114
- Publication Date:
- Research Org.:
- Harvard Univ., Cambridge, MA (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC), Basic Energy Sciences (BES)
- OSTI Identifier:
- 1235135
- Alternate Identifier(s):
- OSTI ID: 1349042
- Grant/Contract Number:
- FG02-09ER16104; SC0001548
- Resource Type:
- Published Article
- Journal Name:
- Proceedings of the National Academy of Sciences of the United States of America
- Additional Journal Information:
- Journal Name: Proceedings of the National Academy of Sciences of the United States of America Journal Volume: 112 Journal Issue: 37; Journal ID: ISSN 0027-8424
- Publisher:
- Proceedings of the National Academy of Sciences
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 60 APPLIED LIFE SCIENCES; cell division; label-free histology; mitotic rate; skin cancer; stimulated Raman scattering microscopy
Citation Formats
Lu, Fa-Ke, Basu, Srinjan, Igras, Vivien, Hoang, Mai P., Ji, Minbiao, Fu, Dan, Holtom, Gary R., Neel, Victor A., Freudiger, Christian W., Fisher, David E., and Xie, X. Sunney. Label-free DNA imaging in vivo with stimulated Raman scattering microscopy. United States: N. p., 2015.
Web. doi:10.1073/pnas.1515121112.
Lu, Fa-Ke, Basu, Srinjan, Igras, Vivien, Hoang, Mai P., Ji, Minbiao, Fu, Dan, Holtom, Gary R., Neel, Victor A., Freudiger, Christian W., Fisher, David E., & Xie, X. Sunney. Label-free DNA imaging in vivo with stimulated Raman scattering microscopy. United States. https://doi.org/10.1073/pnas.1515121112
Lu, Fa-Ke, Basu, Srinjan, Igras, Vivien, Hoang, Mai P., Ji, Minbiao, Fu, Dan, Holtom, Gary R., Neel, Victor A., Freudiger, Christian W., Fisher, David E., and Xie, X. Sunney. Mon .
"Label-free DNA imaging in vivo with stimulated Raman scattering microscopy". United States. https://doi.org/10.1073/pnas.1515121112.
@article{osti_1235135,
title = {Label-free DNA imaging in vivo with stimulated Raman scattering microscopy},
author = {Lu, Fa-Ke and Basu, Srinjan and Igras, Vivien and Hoang, Mai P. and Ji, Minbiao and Fu, Dan and Holtom, Gary R. and Neel, Victor A. and Freudiger, Christian W. and Fisher, David E. and Xie, X. Sunney},
abstractNote = {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. Moreover, 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. In conclusion, 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.},
doi = {10.1073/pnas.1515121112},
journal = {Proceedings of the National Academy of Sciences of the United States of America},
number = 37,
volume = 112,
place = {United States},
year = {Mon Aug 31 00:00:00 EDT 2015},
month = {Mon Aug 31 00:00:00 EDT 2015}
}
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https://doi.org/10.1073/pnas.1515121112
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