scholarly journals Multiphoton Microscopy for Ophthalmic Imaging

2011 ◽  
Vol 2011 ◽  
pp. 1-11 ◽  
Author(s):  
Emily A. Gibson ◽  
Omid Masihzadeh ◽  
Tim C. Lei ◽  
David A. Ammar ◽  
Malik Y. Kahook
Keyword(s):  
Harmonic Generation ◽  
Second Harmonic ◽  
Imaging Techniques ◽  
Multiphoton Microscopy ◽  
Tissue Imaging ◽  
Third Harmonic ◽  
Two Photon ◽  
Disease Mechanisms ◽  
Ophthalmic Imaging ◽  
Anti Stokes

We review multiphoton microscopy (MPM) including two-photon autofluorescence (2PAF), second harmonic generation (SHG), third harmonic generation (THG), fluorescence lifetime (FLIM), and coherent anti-Stokes Raman Scattering (CARS) with relevance to clinical applications in ophthalmology. The different imaging modalities are discussed highlighting the particular strength that each has for functional tissue imaging. MPM is compared with current clinical ophthalmological imaging techniques such as reflectance confocal microscopy, optical coherence tomography, and fluorescence imaging. In addition, we discuss the future prospects for MPM in disease detection and clinical monitoring of disease progression, understanding fundamental disease mechanisms, and real-time monitoring of drug delivery.

scholarly journals Depth resolved label-free multimodal optical imaging platform to study morpho-molecular composition of tissue

2019 ◽  
Vol 18 (5) ◽  
pp. 997-1008 ◽  
Author(s):  
Marco Andreana ◽  
Ryan Sentosa ◽  
Mikael T. Erkkilä ◽  
Wolfgang Drexler ◽  
Angelika Unterhuber
Keyword(s):  
Optical Coherence Tomography ◽  
Raman Scattering ◽  
Second Harmonic Generation ◽  
Harmonic Generation ◽  
Structural Information ◽  
Second Harmonic ◽  
Label Free ◽  
Two Photon ◽  
Non Invasive ◽  
Anti Stokes

The presented multi-modal platform combines optical coherence tomography, two-photon excited fluorescence, second harmonic generation and anti-Stokes Raman scattering to provide molecular and structural information of tissue in a fast and non-invasive manner.

scholarly journals Label-Free Multiphoton Microscopy: Much More Than Fancy Images

2021 ◽  
Vol 22 (5) ◽  
pp. 2657
Author(s):  
Giulia Borile ◽  
Deborah Sandrin ◽  
Andrea Filippi ◽  
Kurt I. Anderson ◽  
Filippo Romanato
Keyword(s):  
Harmonic Generation ◽  
Biomedical Research ◽  
Quantitative Methods ◽  
Second Harmonic ◽  
Multiphoton Microscopy ◽  
Basic Research ◽  
Label Free ◽  
Multiphoton Imaging ◽  
Third Harmonic ◽  
Research Fields

Multiphoton microscopy has recently passed the milestone of its first 30 years of activity in biomedical research. The growing interest around this approach has led to a variety of applications from basic research to clinical practice. Moreover, this technique offers the advantage of label-free multiphoton imaging to analyze samples without staining processes and the need for a dedicated system. Here, we review the state of the art of label-free techniques; then, we focus on two-photon autofluorescence as well as second and third harmonic generation, describing physical and technical characteristics. We summarize some successful applications to a plethora of biomedical research fields and samples, underlying the versatility of this technique. A paragraph is dedicated to an overview of sample preparation, which is a crucial step in every microscopy experiment. Afterwards, we provide a detailed review analysis of the main quantitative methods to extract important information and parameters from acquired images using second harmonic generation. Lastly, we discuss advantages, limitations, and future perspectives in label-free multiphoton microscopy.

scholarly journals MULTIPHOTON MICROSCOPIC IMAGING OF MOUSE INTESTINAL MUCOSA BASED ON TWO-PHOTON EXCITED FLUORESCENCE AND SECOND HARMONIC GENERATION

2013 ◽  
Vol 06 (01) ◽  
pp. 1350004 ◽  
Author(s):  
REN'AN XU ◽  
XIAOQIN ZHU ◽  
NING HE ◽  
SHUANGMU ZHUO ◽  
JIAN XU ◽  
...  
Keyword(s):  
Second Harmonic Generation ◽  
Harmonic Generation ◽  
Second Harmonic ◽  
Multiphoton Microscopy ◽  
Tissue Architecture ◽  
Intestinal Tissue ◽  
Two Photon ◽  
Main Components ◽  
Columnar Cells

Multiphoton microscopy (MPM), based on two-photon excited fluorescence and second harmonic generation, enables direct noninvasive visualization of tissue architecture and cell morphology in live tissues without the administration of exogenous contrast agents. In this paper, we used MPM to image the microstructures of the mucosa in fresh, unfixed, and unstained intestinal tissue of mouse. The morphology and distribution of the main components in mucosa layer such as columnar cells, goblet cells, intestinal glands, and a little collagen fibers were clearly observed in MPM images, and then compared with standard H&E images from paired specimens. Our results indicate that MPM combined with endoscopy and miniaturization probes has the potential application in the clinical diagnosis and in vivo monitoring of early intestinal cancer.

scholarly journals Multiphoton microscopy for label-free multicolor imaging of peripheral nerve

2021 ◽  
Author(s):  
Lars Rishøj ◽  
Iván Coto Hernández ◽  
Siddharth Ramachandran ◽  
Nate Jowett
Keyword(s):  
Peripheral Nerve ◽  
Harmonic Generation ◽  
Second Harmonic ◽  
Yellow Fluorescent Protein ◽  
Multiphoton Microscopy ◽  
Third Harmonic Generation ◽  
Second Harmonic Generation Signal ◽  
Label Free ◽  
Third Harmonic ◽  
Third Harmonic Generation Microscopy

AbstractConventional histomorphometry of peripheral nerve entails lengthy chemical processing, ultrathin sectioning in resin, and imaging by light or electron microscopy. Multiphoton microscopy techniques exist enabling label-free and in vivo imaging of histological samples. Third-harmonic-generation microscopy has recently been demonstrated effective for imaging the myelin sheath of peripheral nerve axons in animal models. Herein, we characterize use of second and third harmonic generation microscopy for label-free imaging of murine and human peripheral nerve via a novel multicolor multiphoton microscope based on a single excitation wavelength at 1300 nm. Second harmonic generation signal from collagen centered about 650 nm delineates neural connective tissue, while third harmonic general signal centered about 433 nm delineates myelin and other lipids. In transgenic mice expressing yellow fluorescent protein linked to the thy1 promoter, three-photon-excitation with emission peak at 527 nm delineates axoplasm. We compare label-free multiphoton imaging of murine and human peripheral nerve against conventional chemical stains and discuss clinical implications of this approach in guiding intraoperative decision making in nerve transfer procedures.

Jadassohn-Pellizzari anetoderma: Study of multiphoton microscopy based on two-photon excited fluorescence and second harmonic generation

2009 ◽  
Vol 19 (6) ◽  
pp. 570-575 ◽  
Author(s):  
Jingjun Zhao ◽  
Jianxin Chen ◽  
Yinghong Yang ◽  
Shuangmu Zhuo ◽  
Xingshan Jiang ◽  
...  
Keyword(s):  
Second Harmonic Generation ◽  
Harmonic Generation ◽  
Second Harmonic ◽  
Multiphoton Microscopy ◽  
Two Photon
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