Multiphoton microscopy for label-free identification of intramural metastasis in human esophageal squamous cell carcinoma

Xu, Jian; Kang, Deyong; Zeng, Yaping; Zhuo, Shuangmu; Zhu, Xiaodong; Jiang, Liwei; Chen, Jianxin; Lin, Jiangbo

doi:10.1364/boe.8.003360

Cited by 23 publications

(10 citation statements)

References 35 publications

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“…The wavelength of SHG from collagen fibers was 405 nm, whereas the TPEF from collagen fibers ranged from 450 to 570 nm. 9,15,22 All of the images were obtained using a laser scanning unit (Fluoview 300, Olympus), a pair of objective lenses for laser focusing and the collection of photons (UPlanSApo 20×/0.75, Olympus), and two photomultiplier tubes for the detection of forward/backward SHG and backward TPEF (R3896, Hamamatsu). In backward imaging mode, SHG and TPEF were split using a dichroic beamsplitter (FF435-Di01, Semrock), and filtered from the intense excitation laser background using a bandpass filter (FF01-405/10, Semrock) and a colored glass (BG39, Schott), respectively.…”

Section: Methodsmentioning

confidence: 99%

“…These limitations have led to the development of advanced optical imaging tools to augment histopathological techniques in cancer research, including confocal microscopy and two-photon excited fluorescence (TPEF)/second harmonic generation (SHG) microscopy. 8,9 Methods which employ these tools show considerable promise in elucidating the microstructure of bio-tissues and in monitoring the structural changes associated with tumor progression. Nevertheless, lots of imaging schemes based on confocal microscopy usually require fluorescent labeling to enhance image contrast, which may undermine the precision in diagnostics, and disturb the way a biological event functions.…”

Section: Introductionmentioning

confidence: 99%

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Label-free characterization of collagen fibers in cancerous esophagus tissues using ratiometric nonlinear optical microscopy

Chen

Lin

et al. 2020

Exp Biol Med (Maywood)

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A quantitative analytical method to discriminate among the various types of cancerous esophagus tissue is essential for accurate cancer staging. This paper reports on the use of ratiometric nonlinear optical microscopy to reveal the ratio of two-photon excited fluorescence (TPEF) to second harmonic generation (SHG) and forward to backward (F/B) SHG from single collagen fibers only in submucosa of esophageal squamous cell carcinoma. This makes it possible to accurately differentiate among the four stages of esophageal cancer, providing results that are in good agreement with histopathology. Furthermore, it is confirmed by polarization-dependent SHG that the varied SHG response in esophageal cancer tissues is mainly from the shrinkage in diameter of collagen fibers instead of the collagen triple helixes altered by cancer cells. Based on the results of TPEF/SHG and F/B SHG ratio, they can cooperatively improve the precision of diagnostics on esophageal cancer and could be transferred to other types of cancer diseases with changed collagen fibers. Impact statement The issue of classifying esophageal cancer at various developmental stages is crucial for determining the optimized treatment protocol for the patients, as well as the prognosis. Precision improvement in staging esophageal cancer keeps seeking quantitative and analytical imaging methods that could augment histopathological techniques. In this work, we used nonlinear optical microscopy for ratiometric analysis on the intrinsic signal of two-photon excited fluorescence (TPEF) and second harmonic generation (SHG) from single collagen fibers only in submucosa of esophageal squamous cell carcinoma (ESCC). The blind tests of TPEF/SHG and forward (F)/backward (B) SHG were demonstrated to compare with the histology conclusion. The discussion of sensitivity and specificity was provided via statistical comparison between the four stages of esophageal cancer. To the best of our knowledge, this is the first study of using these two ratios in combination for staging ESCC.

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Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Label-free characterization of collagen fibers in cancerous esophagus tissues using ratiometric nonlinear optical microscopy

Chen

Lin

et al. 2020

Exp Biol Med (Maywood)

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“…For example, 2PEF microscopy can alone quantify tumor fluorescence of cancer tissues [17]. 2PEF combined with SHG imaging allows the investigation of various biomedical issues, e.g., local invasion at tumor-stromal interface [18,19], basal cell carcinoma [20,21], orientation and polarization dependence of collagen [22][23][24][25], malignant melanoma [26], and squamous cell carcinoma [27]. Besides imaging tumors [28,29], THG is able to measure the nuclear-cytoplasmic (NC) ratio-an important index for scoring skin aging [30].…”

Section: Introductionmentioning

confidence: 99%

Multimodal imaging platform for optical virtual skin biopsy enabled by a fiber-based two-color ultrafast laser source

Chung

Greinert²,

Kärtner

et al. 2019

Biomed. Opt. Express

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We demonstrate multimodal label-free nonlinear optical microscopy in human skin enabled by a fiber-based two-color ultrafast source. Energetic femtosecond pulses at 775 nm and 1250 nm are simultaneously generated by an Er-fiber laser source employing frequency doubling and self-phase modulation enabled spectral selection. The integrated nonlinear optical microscope driven by such a two-color femtosecond source enables the excitation of endogenous two-photon excitation fluorescence, second-harmonic generation, and thirdharmonic generation in human skin. Such a 3-channel imaging platform constitutes a powerful tool for clinical application and optical virtual skin biopsy.

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“…[2][3][4][5] Nonlinear absorption is the origin of several applications in science and technology. Multiphoton absorption processes are often involved in laser-induced damage, 6,7°u orescence microscopy, 8 optical power limiting, 9,10°u orescence up-conversion, 11,12 up-converting lasing 13 and luminescence imaging. 14 One of the experimental methods for characterizing nonlinear absorption is the Z-scan technique.…”

Section: Introductionmentioning

confidence: 99%

Analytical model for a single multiphoton absorption process

Kessi

Hamadouche

2019

J. Nonlinear Optic. Phys. Mat.

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We present a theoretical analysis of a single [Formula: see text]-photon absorption process. We found an approximate analytical but closed-form expression of the normalized transmittance for a thin nonlinear material under an elliptic Gaussian incident beam. The obtained expression can be used to fit the experimental [Formula: see text]-scan traces, allowing the determination of material’s multiphoton absorption coefficient and Rayleigh lengths of the input elliptic Gaussian beam.

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Multiphoton microscopy for label-free identification of intramural metastasis in human esophageal squamous cell carcinoma

Cited by 23 publications

References 35 publications

Label-free characterization of collagen fibers in cancerous esophagus tissues using ratiometric nonlinear optical microscopy

Label-free characterization of collagen fibers in cancerous esophagus tissues using ratiometric nonlinear optical microscopy

Multimodal imaging platform for optical virtual skin biopsy enabled by a fiber-based two-color ultrafast laser source

Analytical model for a single multiphoton absorption process

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