There is a critical clinical need for a noninvasive high-resolution imaging technology for early cancer detection and guidance of biopsy in internal organs. Two-photon fluorescence (TPF) and second harmonic generation (SHG) microscopy is a powerful technology to address the above clinical need by providing structural and biochemical/metabolic information about biological tissues at subcellular resolution without the need for tissue removal or external fluorescent agents. However, its in vivo clinical application remains extremely limited due to the lack of a miniature technology platform. The objective of this multidisciplinary proposal is to develop an all-fiber-optic scanning endomicroscopy technology which is able to bring TPF/SHG microscopy to clinic for internal organ imaging. It involves 5 partners with 2 from academia and 3 from industry. The proposed technology will integrate all essential functions of a scanning laser microscope into a single flexible fiber-optic probe of a small diameter (~2.4-3.4 mm), with built-in mechanisms for femtosecond pulse delivery, dispersion management, nonlinear effect suppression, beam focusing, rapid 2D raster beam scanning, TPF/SHG collection, and focus tracking (or depth scanning). The small size permits its integration with a standard red-flagging technology (such as a gastroscope). In this proposal, we plan to tackle the major challenges in developing such an endomicroscopy technology and evaluate its feasibility for subcellular resolution imaging and for cancer detection.
The Specific Aims are to: (1) Develop new double-clad fibers (DCF) of a pure silica core, large inner clad and numerical aperture to dramatically suppress the in-fiber TPF/SHG background (e.g. by 50 folds) and improve TPF/SHG collection efficiency (e.g. by ~15 folds) over commercially available DCFs; (2) Develop a super-achromatic microlens of a 2.1mm diameter and 0.6 NA to improve the TPF/SHG collection efficiency by at least 20 folds over a GRIN lens; (3) Explore a novel approach based on high-order mode DCFs to suppress nonlinear effects in optic fiber and improve TPF/SHG excitation probability; (4) Develop novel MEMS scanners of a small footprint (1.6 x 1.6 mm) and an extremely low drive voltage (10V max) to achieve rapid 2D raster beam scanning and real-time focus tracking. A fully integrated endomicroscope with customized DCFs, microlens and MEMS scanners will be developed, capable of 3D TPF/SHG imaging; (5) Conduct in vivo endoscopic TPF/SHG imaging of swine esophagus to evaluate the performance, and design, engineering and operation issues of the scanning probe; and (6) Evaluate the feasibility of the proposed technology for cancer detection and tumor margin identification using ex vivo human esophagus specimens, and correlate TPF/SHG endomicroscopy images with corresponding histology. The significance of the proposed research is to translate the powerful TPF/SHG microscopy technology to clinical practice for cancer detection and image-guided biopsy in internal organs, and enable noninvasive real-time visualization of tissue histopathology in situ to significantly improve diagnostic and biopsy yields. In addition, the proposed endomicroscopy technology will also be applicable (although outside the scope of this proposal) to many other clinical scenarios such as for guidance of surgical interventions and for in vivo assessment of metabolic function of living tissues.

Public Health Relevance

The objective of this multidisciplinary proposal is to develop and test an all-fiber-optic 3D scanning endomicroscopy technology, which can bring the powerful bench-top TPF/SHG microscopy technology to clinical practice for imaging internal organs (such as the gastrointestinal tract) that was not previously possible with TPF/SHG microscopy. The proposed technology can function in a form of 'optical biopsy' by providing structural and biochemical/metabolic information about biological tissues at subcellular resolution without the need for tissue removal or external fluorescent agents. Successful completion of the proposed research will bring us a new tool, which enables visualization of histopathology in situ and improves our capability for early cancer detection, tumor margin identification, and guidance of biopsy and interventions.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA153023-05
Application #
8776708
Study Section
Special Emphasis Panel (ZRG1-SBIB-U (55))
Program Officer
Baker, Houston
Project Start
2011-01-20
Project End
2015-11-30
Budget Start
2014-12-01
Budget End
2015-11-30
Support Year
5
Fiscal Year
2015
Total Cost
$420,787
Indirect Cost
$152,247
Name
Johns Hopkins University
Department
Biomedical Engineering
Type
Schools of Medicine
DUNS #
001910777
City
Baltimore
State
MD
Country
United States
Zip Code
21205
Li, Ang; Hall, Gunnsteinn; Chen, Defu et al. (2018) A biopsy-needle compatible varifocal multiphoton rigid probe for depth-resolved optical biopsy. J Biophotonics :e201800229
Li, Kaiyan; Liang, Wenxuan; Mavadia-Shukla, Jessica et al. (2018) Super-achromatic optical coherence tomography capsule for ultrahigh-resolution imaging of esophagus. J Biophotonics :e201800205
Yuan, Wu; Brown, Robert; Mitzner, Wayne et al. (2017) Super-achromatic monolithic microprobe for ultrahigh-resolution endoscopic optical coherence tomography at 800?nm. Nat Commun 8:1531
Hall, Gunnsteinn; Liang, Wenxuan; Li, Xingde (2017) Fitting-free algorithm for efficient quantification of collagen fiber alignment in SHG imaging applications. Biomed Opt Express 8:4609-4620
Park, Hyeon-Cheol; Mavadia-Shukla, Jessica; Yuan, Wu et al. (2017) Broadband rotary joint for high-speed ultrahigh-resolution endoscopic OCT imaging at 800??nm. Opt Lett 42:4978-4981
Liang, Wenxuan; Hall, Gunnsteinn; Messerschmidt, Bernhard et al. (2017) Nonlinear optical endomicroscopy for label-free functional histology in vivo. Light Sci Appl 6:
Yan, Lesan; Li, Xingde (2016) Biodegradable Stimuli-Responsive Polymeric Micelles for Treatment of Malignancy. Curr Pharm Biotechnol 17:227-36
Yuan, Wu; Mavadia-Shukla, Jessica; Xi, Jiefeng et al. (2016) Optimal operational conditions for supercontinuum-based ultrahigh-resolution endoscopic OCT imaging. Opt Lett 41:250-3
Zhang, Xiaoyang; Duan, Can; Liu, Lin et al. (2015) A non-resonant fiber scanner based on an electrothermally-actuated MEMS stage. Sens Actuators A Phys 233:239-245
Zhang, Anqi; Xi, Jiefeng; Liang, Wenxuan et al. (2014) Generic pixel-wise speckle detection in Fourier-domain optical coherence tomography images. Opt Lett 39:4392-5

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