Recent Progress on Strategies and Applications of Imaging for Intestinal Microflora

doi:10.6023/cjoc202112022

Abstract

The microflora in the mammalian gut plays an essential role in maintaining the physiological states and pathological changes of the host, and it is of significance for host health to detect the microflora in the gut. However, the traditional staining methods for bacteria have some drawbacks, such as poor specificity, incompatibility with living cells, and susceptibility, thereby limiting further study on the morphology and function of intestinal microflora. The fluorescence imaging technique could accurately reveal the location of symbiotic, pathogenic, and opportunistic bacteria in the intestine. The fluorescence imaging technique could reflect some information, including bacterial activity, metabolic exchange, and immune interactions between the host and the microbe due to its advantages of non-invasive, less tissue damage, higher specificity, and sensitivity, thereby being widely utilized in bacterial detection. More importantly, the fluorescent-labeled probes are the critical factor in the fluorescence imaging of intestinal microflora, promoting the development of fluorescence imaging technology continuously. This review summarizes the labeling strategies designed for different intestinal microflora, including fluorescent probes and isotope probes, and disscussed the metabolic labeling, the methods of non-metabolic labeling and the specific labeling of metabolites, finally providing a prospect for the development of the fluorescent-labeled probes for intestinal microflora.

Key words: intestinal microflora imaging, fluorescent probes, bioorthogonal reaction, peptidoglycan labeling, D-amino acid metabolic labeling

Cite this article

Na Li, Xiaofeng Tan, Qinglai Yang. Recent Progress on Strategies and Applications of Imaging for Intestinal Microflora[J]. Chinese Journal of Organic Chemistry, 2022, 42(5): 1375-1386.

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Fig. & Tab. 8

Figure 1. Fluorescent-labeled probes for imaging of intestinal microflora based on the metabolic labeling, non-metabolic labeling and specific metabolite labeling methods

Figure 2. (a) Schematic of 8AzKdo characterized by the bioorthogonal reaction to metabolically label G-bacteria (left) and Vanco-Cy3 specifically labelled G+bacteria (right); (b) 8AzKdo and Vanco-Cy3 for dual-color fluorescence imaging of intestinal microflora; (c) Schematic diagram of MOE-BBC method and (d) imaging of ileal symbionts; (e) Three fluorescent probes respectively labeled peptidoglycan, lipopolysaccharide and capsular polysaccharide (a) and (b) Adapted with permission[20], Copyright 2017, American Chemical Society; (c) and (d) Adapted with permission[27], Copyright 2015, Springer Nature; (e) Adapted with permission[28], Copyright 2017, Nature Research

Figure 4. (a) Using non-standard amino acids to track the engineered bacteria in the intestine of mice; (b) bioorthogonal test labelled B. fragilis and (c) PET imaging of it in mice; (d) schematic diagram of ABP-FACS identifying β-glucuronidase-active microbiota in the gut; (e) AzG metabolic labelled intestinal microflora (left) and confocal fluorescence imaging (right) (a) Adapted with permission [38]; Copyright 2018, American Chemical Society; (b) and (c) Adapted with permission[39]; Copyright 2020, Springer; (d) Adapted with permission[43]; Copyright 2018, American Chemical Society; (e) Adapted with permission[44]; Copyright 2020, Oxford University Press

Figure 5. (a) Schematic of a fluorescence STAMP used to track and evaluate the viability of transplanted gut microbiotas; (b) schematic diagram of MeDabLISH analysis strategy; (c) 3D-reconstructed confocal imaging of the bacterial distribution in gut (a) Adapted with permission[36]; Copyright 2019, Nature Research; (b) Adapted with permission[39]; Copyright 2020, Wiley-VCH; (c) Adapted with permission[56]; Copyright 2021, Wiley-VCH

Figure 6. (a) 18F-FDG PET uptake in different colonic regions before and after antibiotic therapy; (b) 13C and 15N labeled threonine metabolically labelled intestinal microflora and FISH images; (c) uptake of FLA-YM and d FLA-RGII by B. theta (a) Adapted with permission[60]; Copyright 2018, Public Library Science; (b) Adapted with permission[64]; Copyright 2013, National Academy of Sciences; (c) Adapted with permission[41]; Copyright 2019, Springer Nature

Figure 7. (a) Schematic diagram of PxB-Cy3 labelled Gram-negative bacteria; (b) application of TriA-TAMRA and Vanco-BODIPY in two-color fluorescence imaging of intestinal microflora in mice; (c) FDGlcU used to evaluate the inhibitory effect of eβG inhibitors on βG activity; (d) PA images of Lactobacillus stained with ICG and E. coli stained with PB (a) Adapted with permission[22]; Copyright 2018, Science Press; (b) Adapted with permission[65]; Copyright 2019, Springer; (c) Adapted with permission[42]; Copyright 2017, Nature Springer; (d) Adapted with permission[66]; Copyright 2017, Optical Society of America

Figure 8. (a) Schematic representation of the BAL assay, and (b) design diagram of carbapenemase-specific fluorescent probes Adapted with permission[67]; Copyright 2021, American Association for the Advancement of Science

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