Multicolored stain-free histopathology with coherent Raman imaging.
Conventional histopathology with hematoxylin & eosin (H&E) has been the gold standard for histopathological diagnosis of a wide range of diseases. However, it is not performed in vivo and requires thin tissue sections obtained after tissue biopsy, which carries risk, particularly in the central nervous system. Here we describe the development of an alternative, multicolored way to visualize tissue in real-time through the use of coherent Raman imaging (CRI), without the use of dyes. CRI relies on intrinsic chemical contrast based on vibrational properties of molecules and intrinsic optical sectioning by nonlinear excitation. We demonstrate that multicolor images originating from CH(2) and CH(3) vibrations of lipids and protein, as well as two-photon absorption of hemoglobin, can be obtained with subcellular resolution from fresh tissue. These stain-free histopathological images show resolutions similar to those obtained by conventional techniques, but do not require tissue fixation, sectioning or staining of the tissue analyzed.
Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA, USA.
Christian W
CW
Freudiger
Rolf
R
Pfannl
Daniel A
DA
Orringer
Brian G
BG
Saar
Minbiao
M
Ji
Qing
Q
Zeng
Linda
L
Ottoboni
Ying
Y
Wei
Wei
W
Ying
Christian
C
Waeber
John R
JR
Sims
Philip L
PL
De Jager
Oren
O
Sagher
Martin A
MA
Philbert
Xiaoyin
X
Xu
Santosh
S
Kesari
X Sunney
XS
Xie
Geoffrey S
GS
Young
D000818:Animals; D001932:Brain Neoplasms / Q000473:pathology; D045744:Cell Line, Tumor; D058948:Cell Tracking / Q000379:methods*; D003711:Demyelinating Diseases / Q000473:pathology; D004195:Disease Models, Animal; D006454:Hemoglobins / Q000737:chemistry; D016591:Histocytological Preparation Techniques; D006801:Humans; D008055:Lipids / Q000737:chemistry; D051379:Mice; D008810:Mice, Inbred C57BL; D008819:Mice, Nude; D011506:Proteins / Q000737:chemistry; D013059:Spectrum Analysis, Raman / Q000379:methods*; D013194:Staining and Labeling; D020521:Stroke / Q000473:pathology; D041623:Tomography, Optical Coherence / Q000295:instrumentation / Q000379:methods*
D006454:Hemoglobins; D008055:Lipids; D011506:Proteins