Nonlinear optical microscopy of the bronchus

Zhuo, Shuangmu; Chen, Jianxin; Yu, Bin; Jiang, Xingshan; Luo, Teng; Liu, Quangang; Chen, Rong; Xie, Shusen

doi:10.1117/1.2982534

Cited by 26 publications

(21 citation statements)

References 45 publications

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“…The approach for determining the spacing between elastic fibers has been described in our previous studies. 4 In this work, the spacing between elastic fibers in normal stroma is 14.13Ϯ 4.12 m ͑n =27 slices of 9 biopsies͒ and in neoplastic stroma is 1.72Ϯ 0.57 m ͑n =27 slices of 9 biopsies͒, indicating that the elastic fibers have a getting together tendency. Second, similar to the measurement of collagen, we also obtained the elastin area.…”

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confidence: 87%

“…3 It is well suited for the observation of unstained samples based on intrinsic sources of nonlinear signals, 4 such as two-photon excited fluorescence ͑TPEF͒ and second-harmonic generation ͑SHG͒. It also has been used for in vivo cancer imaging.…”

mentioning

confidence: 99%

“…4 Briefly, it is an Axiovert 200 microscope ͑Zeiss LSM 510 META͒ equipped with a mode-locked femtosecond Ti:sapphire laser ͑Coherent Mira 900-F͒. The polarization of laser light is the linear polarization.…”

mentioning

confidence: 99%

“…To suppress TPEF signals from collagen, the excitation wavelength at 850 nm was used in this work. 4 An oil immersion objective ͑ϫ63 and NA= 1.4͒ was employed. The META detector was used to detect all signals in backscattered geometry.…”

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confidence: 99%

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Extracting diagnostic stromal organization features based on intrinsic two-photon excited fluorescence and second-harmonic generation signals

et al. 2009

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Abstract. Intrinsic two-photon excited fluorescence ͑TPEF͒ and second-harmonic generation ͑SHG͒ signals are shown to differentiate between normal and neoplastic human esophageal stroma. It was found that TPEF and SHG signals from normal and neoplastic stroma exhibit different organization features, providing quantitative information about the biomorphology and biochemistry of tissue. By comparing normal with neoplastic stroma, there were significant differences in collagen-related changes, elastinrelated changes, and alteration in proportions of matrix molecules, giving insight into the stromal changes associated with cancer progression and providing substantial potential to be applied in vivo to the clinical diagnosis of epithelial precancers and cancers. A number of optical spectroscopic and imaging techniques have widely been used for the in vivo, real-time detection of epithelial precancers and cancers.1 However, most studies focus only on the epithelium to analyze morphological, structural, and architectural changes that accompany development of epithelial precancers.2 Recently, it has been recognized that stromal biology is also altered significantly with various pathological processes.2 However, there is a lack of utilizing the alterations in the stroma as an intrinsic indicator of disease states, which can extract quantitative information about the biomorphology and biochemistry and can provide a new means to improve early detection of neoplastic changes.Multiphoton microscopy has several advantages over traditional confocal microscopy, providing high-resolution images at increased imaging depths, minimal out-of-plane absorption, and inherent optical sectioning.3 It is well suited for the observation of unstained samples based on intrinsic sources of nonlinear signals, 4 such as two-photon excited fluorescence ͑TPEF͒ and second-harmonic generation ͑SHG͒. It also has been used for in vivo cancer imaging. 3Our primary goals in this study are to investigate the intrinsic TPEF and SHG signals in epithelial stroma and to extract biomorphologic and biochemical features for potential tissue diagnosis.The multiphoton system used in this study has been described previously. 4 Briefly, it is an Axiovert 200 microscope ͑Zeiss LSM 510 META͒ equipped with a mode-locked femtosecond Ti:sapphire laser ͑Coherent Mira 900-F͒. The polarization of laser light is the linear polarization. To suppress TPEF signals from collagen, the excitation wavelength at 850 nm was used in this work. 4 An oil immersion objective ͑ϫ63 and NA= 1.4͒ was employed. The META detector was used to detect all signals in backscattered geometry. The average laser power at the sample was 5 mW. In this study, all images were 512ϫ 512 pixels. The images were obtained at 2.56 s per pixel. In this work, considering that the surface of the sections is not absolutely flat and that the signals in the surface of sections is not absolutely maximal, we defined a depth of 0 m, referring to the position where the multiphoton signals of reflection from the interfac...

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confidence: 87%

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confidence: 99%

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confidence: 99%

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confidence: 99%

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Extracting diagnostic stromal organization features based on intrinsic two-photon excited fluorescence and second-harmonic generation signals

et al. 2009

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“…In addition, it has been reported that the ratio of NADH over FAD°uorescence calculated by the equation, FAD 535 /(FAD 535 þNADH 475 Þ, is a parameter related to cellular metabolism. 15,16 In this study, the ratio of NADH over FAD°uorescence in ROI 1 region was 1:81 AE 0:03, which re°ected the metabolic state of chondrocytes and may be found application in characterization of pathological changes for distinguishing abnormal elastic cartilage from an intact one.…”

Section: Depth-resolved Spectral Imaging and Emission Spectra Of Elasmentioning

confidence: 91%

Nonlinear Spectral Imaging of Elastic Cartilage in Rabbit Ears

Chen

Guo

et al. 2013

J. Innov. Opt. Health Sci.

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Elastic cartilage in the rabbit external ear is an important animal model with attractive potential value for researching the physiological and pathological states of cartilages especially during wound healing. In this work, nonlinear optical microscopy based on two-photon excited°uor-escence and second harmonic generation were employed for imaging and quantifying the intact elastic cartilage. The morphology and distribution of main components in elastic cartilage including cartilage cells, collagen and elastic¯bers were clearly observed from the high-resolution two-dimensional nonlinear optical images. The areas of cell nuclei, a parameter related to the pathological changes of normal or abnormal elastic cartilage, can be easily quanti¯ed. Moreover, the three-dimensional structure of chondrocytes and matrix were displayed by constructing three-dimensional image of cartilage tissue. At last, the emission spectra from cartilage were obtained and analyzed. We found that the di®erent ratio of collagen over elastic¯bers can be used to locate the observed position in the elastic cartilage. The redox ratio based on the ratio of nicotinamide adenine dinucleotide (NADH) over°avin adenine dinucleotide (FAD)°uorescence can also be calculated to analyze the metabolic state of chondrocytes in di®erent regions. Our results demonstrated that this technique has the potential to provide more accurate and comprehensive information for the physiological states of elastic cartilage.

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Review of optical methods for noninvasive imaging of skin fibroblasts—From in vitro to ex vivo and in vivo visualization

Nikolaev,

Kistenev,

Kröger

et al. 2024

Journal of Biophotonics

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Fibroblasts are among the most common cell types in the stroma responsible for creating and maintaining the structural organization of the extracellular matrix (ECM) in the dermis, skin regeneration, and a range of immune responses. Until now, the processes of fibroblast adaptation and functioning in a varying environment have not been fully understood. Modern laser microscopes are capable of studying fibroblasts in vitro and ex vivo. One‐photon‐ and two‐photon‐excited fluorescence microscopy, Raman spectroscopy/micro‐spectroscopy are well‐suited non‐invasive optical methods for fibroblast imaging in vitro and ex vivo. In vivo staining‐free fibroblast imaging is not still implemented. The exception is fibroblast imaging in tattooed skin. Although in vivo non‐invasive staining‐free imaging of fibroblasts in the skin has not yet been implemented, it is expected in the future. This review summarizes the state of the art in fibroblast visualization using optical methods and discusses the advantages, limitations, and prospects for future non‐invasive imaging.This article is protected by copyright. All rights reserved.

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Nonlinear optical microscopy of the bronchus

Cited by 26 publications

References 45 publications

Extracting diagnostic stromal organization features based on intrinsic two-photon excited fluorescence and second-harmonic generation signals

Extracting diagnostic stromal organization features based on intrinsic two-photon excited fluorescence and second-harmonic generation signals

Nonlinear Spectral Imaging of Elastic Cartilage in Rabbit Ears

Review of optical methods for noninvasive imaging of skin fibroblasts—From in vitro to ex vivo and in vivo visualization

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