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26BIOMEDICAL APPLICATIONS OF NONLINEAR OPTICAL MICROIMAGING TECHNIQUES
SHIQI WANG1, FANGRUI LIN1, SHENG REN1, YIHUA ZHAO1AND LIWEI LIU1*
1Key Laboratory of Optoelectronic Devices and Systems of Ministry of Education & Guangdong Province, College of Physics and
Optoelectronic Engineering, Shenzhen University, China.
*E-mail: [email protected]
Abstract
Optical microscopy imaging technology is a research hotspot in the field of biomedical photonics. In the past two decades, optical microscopy imaging technology has developed very rapidly, constantly breaking through the traditional limits, and increasingly used in the research on subcellular, cellular, tissue and in vivo imaging research, providing a unique perspective for real-time dynamic observation of life systems and solving many key scientific issues in the field of "biomedical photonics".
Our research group has carried out many years of basic application research of nonlinear optical microscopy imaging technology in biomedicine, covering Two-photon Excitation Fluorescence(TPF), Second-Harmonic Generation(SHG), and Stimulated Raman Scattering(SRS) at different levels of molecules, cells, tissues, and living bodies. These optical imaging technologies provide powerful technical support for the monitoring of the microenvironment during tumor treatment. With the help of mature optical microscopic imaging technology, the clinical progress of qualitative and quantitative analysis of the microenvironment during tumor treatment can be promoted.
Tumor microenvironment refers to the internal and external environment in which tumors occur, grow and metastasize. The relationship between tumor microenvironment and tumor is called "seed and soil". In the process of tumor diagnosis and treatment, tumor microenvironment plays a vital role. Therefore, imaging monitoring of the tumor microenvironment has become an assessment method in the process of tumor treatment.
This report mainly introduces the research work of non-linear optical microscopy imaging technology in tumor microenvironment imaging carried out by our team recently, including tumor metastasis path, intracellular calcium ion regulation, etc.
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i = 0s t = 6s t=12s t = 19 s t = 25 s
-
? 40. ¡T 20 E o < 0 2 3 4 5 - 1
7 13 1 9 26 ; 12 38 45 51 58 64 70 77 83 90 E
Stimulation ■ Reference Background
w r -
*= 0s -i = 6s -1=125 .+=19 s -4 = 25 5 t=.96s
SQ + Rhod-2 ■ *
200 1001
13 19 26 32 38 45 51 50 64 70 77 83 90 96
Stimulation Reference Background
Figure 1:Specifific laser and dye were required for LIDSICA. (a) Cells in a Petri dish were stimulated by a 640 nm laser, and the Ca2+ levels were indicated by a 488 nm (for Fluo-4) or 561 nm (for Rhod-2) laser via a 60x objective. (b, c) OV3 cells stimulated 15x in 100 s showed a Ca2+ rise of ~37% (green, indicated by Fluo-4) (b) or of ~155%(yellow, indicated by Rhod-2 in another cell) (c).
TIIG Heterogeneity SHG Collagen ___.....: I(3)PA FAD
gMMb ■ %
Figure2: Comparison between stain-free MAMG microscopy and H&E histopathology in a cancer colony followed by cancer-
associated angiogenesis (star) as backup.
References
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[2] Binglin Shen, Shiqi Wang, Ganapathi Bharathi, Yangping Li, Fangrui Lin, Rui Hu, Liwei Liu, Junle Qu, Rapid and Targeted Photoactivation of Ca2+ ChannelsMediated by Squaraine To Regulate Intracellular andIntercellular Signaling Processes, Anal. Chem., 8497-8505, 2020.
[3] Binglin Shen, Liwei Liu, Junshuai Yan, Shiqi Wang, Feifan Zhou, Yihua Zhao, Rui Hu, Junle Qu, Large-field stain-free histopathology of metastatic colonization by multimodal optical methods,Theranostics 2020, Vol. 10, Issue 4.
Abstract Graphic
