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2007•Journal of Biomedical OpticsOpen access

Multiphoton fluorescence and second harmonic generation microscopy for imaging infectious keratitis

Hsin‐Yuan Tan, Yen Sun, Wen Lo, Shu‐Wen Teng, Ruei-Jr Wu, Shiou‐Hwa Jee, Wei‐Chou Lin, Ching‐Hsi Hsiao, Hsin‐Chiung Lin, Yeong-Fong Chen, David Hui‐Kang, Samuel Chao‐Ming Huang, Sung‐Jan Lin, Chen‐Yuan Dong

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Abstract

The purpose of this study is to demonstrate the application of multiphoton fluorescence and second harmonic generation (SHG) microscopy for the ex-vivo visualization of human corneal morphological alterations due to infectious processes. The structural alterations of both cellular and collagenous components can be respectively demonstrated using fluorescence and SHG imaging. In addition, pathogens with fluorescence may be identified within turbid specimens. Our results show that multiphoton microscopy is effective for identifying structural alterations due to corneal infections without the need of histological processing. With additional developments, multiphoton microscopy has the potential to be developed into an imaging technique effective in the clinical diagnosis and monitoring of corneal infections.

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What this paper is about

The purpose of this study is to demonstrate the application of multiphoton fluorescence and second harmonic generation (SHG) microscopy for the ex-vivo visualization of human corneal morphological alterations due to infectious processes. The structural alterations of both cellular and collagenous components can be respectively demonstrated using fluorescence and SHG imaging. In addition, pathogens with fluorescence may be identified within turbid specimens. Our results show that multiphoton microscopy is effective for identifying structural alterations due to corneal infections without the need of histological processing. With additional developments, multiphoton microscopy has the potential to be developed into an imaging technique effective in the clinical diagnosis and monitoring of corneal infections.

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Available abstract

The purpose of this study is to demonstrate the application of multiphoton fluorescence and second harmonic generation (SHG) microscopy for the ex-vivo visualization of human corneal morphological alterations due to infectious processes. The structural alterations of both cellular and collagenous components can be respectively demonstrated using fluorescence and SHG imaging. In addition, pathogens with fluorescence may be identified within turbid specimens. Our results show that multiphoton microscopy is effective for identifying structural alterations due to corneal infections without the need of histological processing. With additional developments, multiphoton microscopy has the potential to be developed into an imaging technique effective in the clinical diagnosis and monitoring of corneal infections.

Key concepts: Microscopy, Multiphoton fluorescence microscope, Fluorescence microscope, Fluorescence-lifetime imaging microscopy, Second-harmonic generation, Fluorescence, Two-photon excitation microscopy, Materials science

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