Ex vivo imaging and quantification of liver fibrosis using second-harmonic generation microscopy.
Conventionally, liver fibrosis is diagnosed using histopathological techniques. The traditional method is time-consuming in that the specimen preparation procedure requires sample fixation, slicing, and labeling. Our goal is to apply multiphoton microscopy to efficiently image and quantitatively analyze liver fibrosis specimens bypassing steps required in histological preparation. In this work, the combined imaging modality of multiphoton autofluorescence (MAF) and second-harmonic generation (SHG) was used for the qualitative imaging of liver fibrosis of different METAVIR grades under label-free, ex vivo conditions. We found that while MAF is effective in identifying cellular architecture in the liver specimens, it is the spectrally distinct SHG signal that allows the characterization of the extent of fibrosis. We found that qualitative SHG imaging can be used for the effective identification of the associated features of liver fibrosis specimens graded METAVIR 0 to 4. In addition, we attempted to associate quantitative SHG signal to the different METAVIR grades and found that an objective determination of the extent of disease progression can be made. Our approach demonstrates the potential of using multiphoton imaging in rapid classification of ex vivo liver fibrosis in the clinical setting and investigation of liver fibrosis-associated physiopathology in animal models in vivo.
Sun
Tzu-Lin
TL
National Taiwan University, Department of Physics, Taipei 10617, Taiwan.
Liu
Yuan
Y
Sung
Ming-Chin
MC
Chen
Hsiao-Ching
HC
Yang
Chun-Hui
CH
Hovhannisyan
Vladimir
V
Lin
Wei-Chou
WC
Jeng
Yung-Ming
YM
Chen
Wei-Liang
WL
Chiou
Ling-Ling
LL
Huang
Guan-Tarn
GT
Kim
Ki-Hean
KH
So
Peter T C
PT
Chen
Yang-Fang
YF
Lee
Hsuan-Shu
HS
Dong
Chen-Yuan
CY
D000704:Analysis of Variance; D018450:Disease Progression; D006801:Humans; D007150:Immunohistochemistry; D008103:Liver Cirrhosis / Q000175:diagnosis / Q000473:pathology*; D033401:Microscopy, Acoustic / Q000379:methods*; D036641:Microscopy, Fluorescence, Multiphoton / Q000379:methods*; D012815:Signal Processing, Computer-Assisted