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This content will become publicly available on March 15, 2016

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

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 » chromatin dynamic studies and label-free optical histology in real time.« less
Authors:
 [1] ;  [1] ;  [2] ;  [2] ;  [1] ;  [1] ;  [1] ;  [2] ;  [1] ;  [2] ;  [1]
  1. Harvard Univ., Cambridge, MA (United States)
  2. Brigham and Women's Hospital (Harvard Medical School), Boston, MA (United States)
Publication Date:
OSTI Identifier:
1235135
Grant/Contract Number:
SC0001548; FG02-09ER16104
Type:
Published Article
Journal Name:
Proceedings of the National Academy of Sciences of the United States of America
Additional Journal Information:
Journal Volume: 112; Journal Issue: 37; Journal ID: ISSN 0027-8424
Publisher:
National Academy of Sciences, Washington, DC (United States)
Research Org:
Harvard Univ., Cambridge, MA (United States)
Sponsoring Org:
USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22)
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