|MD1||Utilizing nonlinear optical microscopy to investigate the development of early cancer in nude mice in vivo|
Chun-Chin Wang1, Feng-Chieh Li1, Sung-Jan Lin2, Wen Lo1, Chen-Yuan Dong1
1Department of Physics, National Taiwan University.
In this investigation, we used nonlinear optical microscopy to image and analyze normal and carcinogen DMBA treated skin
tissues of nude mice in vivo. Using the results obtained from two-photon autofluroescence and second harmonic generation
(SHG) images and the application of ASI (Autofluorescence versus SHG Index), we can visualize the interaction between
mouse skin cells and connective tissue.
|MD2||Multiphoton Fluorescence Diagnosis of Hepatocellular Carcinoma|
Tzu-Lin Sun1,Yuan Liu1, Ming-Chin Sung1, Chun-Hui Yang1,
1Department of Physics National Taiwan University,
Hepatic disease has always been a major health problem in Taiwan, and hepatocelular carcinoma (HCC) is among the leading
causes of death. Although doctors diagnose the degree of liver carcinoma according to their clinical experience, there is
a need for an objective diagnosis of the extent of this cancer. In this work, we attempt to develop an objective
criterion to help improving the accuracy of HCC classification which will be a useful aid for clinical investigations.
|MD3||The effects of depilatory agents as a penetration enhancer on human stratum corneum structures|
Jin-Ning Lee 1, Shiou-Hwa Jee 2, Chih-Chieh Chan 2, Wen Lo 1, Sung-Jan Lin 1,3 , Chen-Yuan Dong 1
1 Department of Physics, National Taiwan University, Taipei, Taiwan;
The aim of this research is to characterize the penetration enhancing effect of depilatory, and to investigate the change of stratum corneum(SC). Human foreskin was treated by a depilatory for 10 minutes and then using fluorescent probe, hydrophilic sulforhodamine (SRB), to model drugs penetration that was quantified by twophoton microscopy. The structural alternations of SC are assessed by routine histology, transmission electron microscopy and Nile red staining. The results showed that the penetration of fluorescent probe can be enhanced. Nile red staining revealed, instead of a regular lipid layers around the brick of corneocytes, a disorganized and homogenized pattern of lipid distribution. Ultrastructural analysis showed that the protein envelope of coenocytes disintegrated, especially intercellular lipid of SC. We concluded that depilatory enhance drug penetration by disrupting both the cellular integrity of corneocytes and the regular packing of intercellular lipid of SC.
|MD4||Intravital multiphoton microscopy for imaging hepatobiliary function|
Feng-Chien Li1 , Tzu-Lin Sun 1, Shu-Mei Yang 2, Hsuan-Shu Lee 2, Chen-Yuan Dong 1
1 Department of Physics, National Taiwan University, Taipei 106, Taiwan;
Liver is the chemical factory in body responsible for important functions such as metabolism and detoxification. When liver can not be regenerated in time to amend damages that has occurred, failure of hepatic functions such as liver failure and metabolic disease can result. Traditionally, the study of liver pathology has depended on histological techniques, but such methods are limited to ex-vivo observation. In order to study hepatic metabolism in vivo, we have designed a hepatic imaging chamber made of biocompatible titanium alloy (6V4Al-Ti, ELI grade). In combination with multiphoton and second harmonic generation microscopy, our approach allows the intravital observation of hepatic intravital activities to be achieved. Processes such as hepatic metabolism and disease progression can be studied using this methodology.
|MD5||The Observation of Interaction between Pseudomonas aeruginosa and Cornea by the Use of Two-Photon Fluorescence and Second Harmonic Generation Microscopy|
Yu Lin Chang1 Wen Lo1 Hsin Yuan Tan2 Chen Yuan Dong1
1Department Of Physics, National Taiwan University , Taipei 106, Taiwan,
As the contact lens becomes popular, the issue of associated corneal infection becomes increasingly important in ophthalmology. Pseudomonas aeruginosa is a bacteria species which occurs frequently in infected corneas. Using minimally invasive, two-photon fluorescence and second harmonic generation microscopy, the structural alteration to collagen fibrils of corneal stroma can be investigated. We acquired multi-photon images on infected corneal button specimens at different times following the introduction of pathogens. Our results show that collagen destruction becomes increasingly more significant with time. We hope that this approach can eventually be used for the clinical diagnosis of the infectious corneas without destructive biopsy procedures.
|MD6||Observation of the influence of force on collagen production by chondrocytes using two-photon and second harmonic generation.|
Chia-Cheng Chang1,Hsuan-Shu Lee2.3,Ling-Ling Chiou3,Chi-Hsiu Huang1,Wen Lo1,Chen-Yan Dong1.
1National Taiwan University Department of Physics Taipei 106 Taiwan
We cultivate mesenchymal stem cells (MSCs) in a medium with TGF-£]3 added. TGF-£]3 induce the MSCs to differentiate into chomdrocytes and the chondrocytes subsequently produces collagen. During culture, we impart force on the cells by using a centrifuge. We choose three forces-0G , 10000G , 20000G and exerted the force on the cells for 15 minutes a day for 4 weeks. We observed that the cells exerted with greater force produce collagen that tend to accumulate in lumps at early stage, and generate stronger second harmonic signal.
|MD7||Imaging Human Bone Marrow Mesenchymal Stem Cell Morphogenesis in Chitosan Scaffold by Multiphoton Microscopy|
Chi-Hsiu Huang 1, Hsuan-Shu Lee 2,3, Ling-Ling Chiou 3, Chia-Cheng Chang 1, Wen Lo 1, Chen-Yuan Dong 1
1 National Taiwan University Department of Physics Taipei 106 Taiwan;
One important goal in tissue engineering is cultiviating cartilage tissue for transplantation. The most common method to cultivate cartilage is to seed human bone marrow mesenchymal stem cells on three dimensional biocompatiable and biodegradable scaffold materials,then add the signal factor TGF£]-3 to induce the cells to produce collagen that would grow into cartilage tissue.In this study,we seeded mesenchymal stem cells on a chitosan scaffold and added TGF£]-3 to induce cartilage tissue producing and image the process using multiphoton and second harmonic generation microscopy.
|MD8||Structural Alteration of Corneal Scar Tissue Revealed by Multiphoton Microscopy|
Wen Lo 1,Chiu-Mei Hsueh 1, Shu-Wen Teng 1,Yen Sun 1, Sung-Jan Lin 2,3, Ching-Hsi Hsiao 4,5, Wei-Chou Lin 6, Hsin-Yuan Tan 2,4,5,
1 Department of Physics, National Taiwan University, Taipei 106, Taiwan,
The special arrangement of collagen fibers in corneal stroma provides extraordinary high transparency of cornea. However, wound healing process after clinical treatments, such as refractive surgeries, can disrupt collagen alignment and hinder corneal transparency. While works have been focused on the mechanism, the availability of a non-invasive in vivo imaging technique may be of valuable importance for investigating this complicated physiological response. Here we demonstrate the multiphoton fluorescence and second harmonic generation (SHG) ex vivo imaging of a full-thickness corneal scar tissue ten years following wounding. We intend to demonstrate the structural alterations to the cornea following wound healing process and the potential of application of this novel optical technique as a clinical diagnostic and monitoring tool for corneal pathologies.
|MD9||High resolution multiphoton imaging of acute liver inflammation using hepatic imaging chamber|
Tzu-Lin Sun1,Feng-Chieh Li1,Shu-Mei Yang3,Wen-Pu Hong1,
1Department of Physics National Taiwan University,
Conventional hepatic research relies heavily on histological images for morphological information of liver tissue. Static
histological images, however, can not provide real-time dynamic information of in vivo physiological processes such as
cell motion. In this work, we developed an intravital imaging system to study the effects of acute inflammation of liver
on immune system reaction. We hope that this methodology can be used to reveal in vivo details of acute inflammation of
liver and allow us to better understand hepatic processes.
|MD10||Ex vivo optical biopsy of eyes by using mulitphoton microscopy|
Wen Lo1, Shu-Wen Teng1, Hsin-Yuan Tan2,3, Hsiao-Ching Chen9, Hsuan-Shu Lee9, Yang-Fan Chen1, Ju-Li Peng4, Huei-Hsing Lin4, Ki Hean Kim5, Yen Sun1, Wei-Chou Lin6, Sung-Jan Lin3,7, Shiou-Hwa Jee7,8, Peter T. C. So5, Chen-Yuan Dong1
1Microscopic Biophysics Laboratory, Department of Physics, National Taiwan University, Taipei 106, Taiwan
The aim of this work is to demonstrate that multiphoton microscopy is a preferred technique to obtain spectrally resolved
morphologic features of the cornea, limbus, conjunctiva, and sclera in ex vivo eyes without slicing and staining. At the
micron resolution, multiphoton imaging can provide both large histological features and detailed structure of epithelium,
corneal collagen fibril bundles and keratocytes.