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Label-free whole-colony imaging and metabolic analysis of metastatic pancreatic cancer by an autoregulating flexible optical system.

ABSTRACT: Cancer metastasis is a Gordian knot for tumor diagnosis and therapy. Many studies have demonstrated that metastatic processes are inevitably affected by the tumor microenvironment. Histopathology is used universally as the gold standard for cancer diagnosis despite the lengthy preparation process and invasiveness. Methods: Here, we introduced a supercontinuum and super-wide-tuning integrated multimodal platform, which combines the confocal, nonlinear and fluorescence lifetime microscopy with autoregulations, for label-free evaluation of fresh tissue and pathological sections. Based on various automated tunable lasers, synchronized and self-adjusting components and eight fast switching detection channels, the system features fast, large-field and subcellular-scale imaging of exogenous and endogenous fluorophores, nonlinear coherent scattering and lifetime contrast. Results: With such an integrated multi-dimensional system, we searched the metastatic region by two-photon and three-photon excited autofluorescence, analyzed the cancer invasion by second harmonic generation and revealed the affected cellular metabolism by phasor-lifetime. We demonstrated the flexible measurement of multiple nonlinear modalities at NIR I and II excitation with a pre-compensation for group delay dispersion of ~7,000 fs2 and low power of <40 mW, and of dual autofluorescence lifetime decays for phasor approach to decompose cancer-associated and disassociated components. This significantly revealed the metastatic and metabolic optical signatures of the whole colony of pancreatic cancers. Conclusion: The synergistic effect of the system demonstrates the great potential to translate this technique into routine clinical applications, particularly for large-scale and quantitative studies of metastatic colonization.

SUBMITTER: Shen B

PROVIDER: S-EPMC6993220 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature

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Label-free whole-colony imaging and metabolic analysis of metastatic pancreatic cancer by an autoregulating flexible optical system.

Shen Binglin B Yan Junshuai J Wang Shiqi S Zhou Feifan F Zhao Yihua Y Hu Rui R Qu Junle J Liu Liwei L

Theranostics 20200101 4

Cancer metastasis is a Gordian knot for tumor diagnosis and therapy. Many studies have demonstrated that metastatic processes are inevitably affected by the tumor microenvironment. Histopathology is used universally as the gold standard for cancer diagnosis despite the lengthy preparation process and invasiveness. <b>Methods</b>: Here, we introduced a supercontinuum and super-wide-tuning integrated multimodal platform, which combines the confocal, nonlinear and fluorescence lifetime microscopy w ...[more]

PMID: 32042340

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Project description:UNLABELLED:We investigated the application capabilities of a laser optical sensor, BARDOT (bacterial rapid detection using optical scatter technology) to generate differentiating scatter patterns for the 20 most frequently reported serovars of Salmonella enterica. Initially, the study tested the classification ability of BARDOT by using six Salmonella serovars grown on brain heart infusion, brilliant green, xylose lysine deoxycholate, and xylose lysine tergitol 4 (XLT4) agar plates. Highly accurate discrimination (95.9%) was obtained by using scatter signatures collected from colonies grown on XLT4. Further verification used a total of 36 serovars (the top 20 plus 16) comprising 123 strains with classification precision levels of 88 to 100%. The similarities between the optical phenotypes of strains analyzed by BARDOT were in general agreement with the genotypes analyzed by pulsed-field gel electrophoresis (PFGE). BARDOT was evaluated for the real-time detection and identification of Salmonella colonies grown from inoculated (1.2 × 10(2) CFU/30 g) peanut butter, chicken breast, and spinach or from naturally contaminated meat. After a sequential enrichment in buffered peptone water and modified Rappaport Vassiliadis broth for 4 h each, followed by growth on XLT4 (~16 h), BARDOT detected S. Typhimurium with 84% accuracy in 24 h, returning results comparable to those of the USDA Food Safety and Inspection Service method, which requires ~72 h. BARDOT also detected Salmonella (90 to 100% accuracy) in the presence of background microbiota from naturally contaminated meat, verified by 16S rRNA sequencing and PFGE. Prolonged residence (28 days) of Salmonella in peanut butter did not affect the bacterial ability to form colonies with consistent optical phenotypes. This study shows BARDOT's potential for nondestructive and high-throughput detection of Salmonella in food samples. IMPORTANCE:High-throughput screening of food products for pathogens would have a significant impact on the reduction of food-borne hazards. A laser optical sensor was developed to screen pathogen colonies on an agar plate instantly without damaging the colonies; this method aids in early pathogen detection by the classical microbiological culture-based method. Here we demonstrate that this sensor was able to detect the 36 Salmonella serovars tested, including the top 20 serovars, and to identify isolates of the top 8 Salmonella serovars. Furthermore, it can detect Salmonella in food samples in the presence of background microbiota in 24 h, whereas the standard USDA Food Safety and Inspection Service method requires about 72 h.

Dataset Information

Label-free whole-colony imaging and metabolic analysis of metastatic pancreatic cancer by an autoregulating flexible optical system.

Publications

Label-free whole-colony imaging and metabolic analysis of metastatic pancreatic cancer by an autoregulating flexible optical system.

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