Molecular third-harmonic-generation microscopy through resonance enhancement with absorbing dye.
This paper reports a facile and effortless method to realize three-dimensional (3D) molecular third-harmonic-generation (THG) microscopy through the technique of resonance enhancement with absorbing dye. Hematoxylin, a popular absorbing stain, is applied as an example to verify the multiphoton resonant enhancement based on the 1230 nm excitation light and can selectively enhance THG yield at cell nuclear sites in the studied specimens, serving as a cell nucleus contrast agent. It is concluded that combining THG microscopy with the mature staining technique can readily achieve 3D molecular imaging without using fluorescence.
Yu
Che-Hang
CH
Department of Electrical Engineering and Graduate Institute of Photonics and Optoelectronics, National Taiwan University, Taipei 10617, Taiwan.
Tai
Shih-Peng
SP
Kung
Chun-Ta
CT
Lee
Wen-Jeng
WJ
Chan
Yi-Fan
YF
Liu
Hsiang-Lin
HL
Lyu
Jiun-Yi
JY
Sun
Chi-Kuang
CK
D002478:Cells, Cultured; D004396:Coloring Agents; D006416:Hematoxylin; D006801:Humans; D007089:Image Enhancement / Q000379:methods*; D007090:Image Interpretation, Computer-Assisted / Q000379:methods*; D021621:Imaging, Three-Dimensional / Q000379:methods*; D008168:Lung / Q000473:pathology; D008175:Lung Neoplasms / Q000473:pathology*; D008853:Microscopy / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D020349:Surface Plasmon Resonance / Q000379:methods*
D004396:Coloring Agents; D006416:Hematoxylin