PMID- 34309601
OWN - NLM
STAT- MEDLINE
DCOM- 20211015
LR  - 20211015
IS  - 1940-087X (Electronic)
IS  - 1940-087X (Linking)
IP  - 173
DP  - 2021 Jul 9
TI  - Visualization of Gut Microbiota-host Interactions via Fluorescence In Situ 
      Hybridization, Lectin Staining, and Imaging.
LID - 10.3791/62646 [doi]
AB  - Measuring the localization of microbes within their in vivo context is an 
      essential step in revealing the functional relationships between the microbiota 
      and the vertebrate gut. The spatial landscape of the gut microbiota is tightly 
      controlled by physical features - intestinal mucus, crypts, and folds - and is 
      affected by host-controlled properties such as pH, oxygen availability, and 
      immune factors. These properties limit the ability of commensal microbes and 
      pathogens alike to colonize the gut stably. At the micron-scale, microbial 
      organization determines the close-range interactions between different microbes 
      as well as the interactions between microbes and their host. These interactions 
      then affect large-scale organ function and host health. This protocol enables the 
      visualization of the gut microbiota spatial organization from distances between 
      cells to organ-wide scales. The method is based on fixing gut tissues while 
      preserving intestinal structure and mucus properties. The fixed samples are then 
      embedded, sectioned, and stained to highlight specific bacterial species through 
      fluorescence in situ hybridization (FISH). Host features, such as mucus and host 
      cell components, are labeled with fluorescently labeled lectins. Finally, the 
      stained sections are imaged using a confocal microscope utilizing tile-scan 
      imaging at high magnification to bridge the micron to centimeter length scales. 
      This type of imaging can be applied to intestinal sections from animal models and 
      biopsies from human tissues to determine the biogeography of the microbiota in 
      the gut in health and disease.
FAU - Ng, Katharine M
AU  - Ng KM
AD  - School of Biomedical Engineering, University of British Columbia; Department of 
      Microbiology and Immunology, University of British Columbia.
FAU - Tropini, Carolina
AU  - Tropini C
AD  - School of Biomedical Engineering, University of British Columbia; Department of 
      Microbiology and Immunology, University of British Columbia; Humans and the 
      Microbiome Program, Canadian Institute for Advanced Research (CIFAR); 
      carolina.tropini@ubc.ca.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Video-Audio Media
DEP - 20210709
PL  - United States
TA  - J Vis Exp
JT  - Journal of visualized experiments : JoVE
JID - 101313252
RN  - 0 (Lectins)
SB  - IM
MH  - Animals
MH  - *Gastrointestinal Microbiome
MH  - Humans
MH  - In Situ Hybridization, Fluorescence
MH  - Lectins
MH  - *Microbiota
MH  - Mucus
EDAT- 2021/07/27 06:00
MHDA- 2021/10/16 06:00
CRDT- 2021/07/26 12:25
PHST- 2021/07/26 12:25 [entrez]
PHST- 2021/07/27 06:00 [pubmed]
PHST- 2021/10/16 06:00 [medline]
AID - 10.3791/62646 [doi]
PST - epublish
SO  - J Vis Exp. 2021 Jul 9;(173). doi: 10.3791/62646.