eoe sub articles

EoE Sub Articles

All procedures involving sample collection have been performed in accordance with the institute&#39;s IRB guidelines.
1. Tissue Preparation for Fixation and Paraffin Embedding
NOTE:&#160;Biopsies were taken from consenting women undergoing cystoscopy with electro-fulguration of trigonitis for the advanced management of recurrent urinary tract infection (rUTI). rUTI is defined as 3 UTIs in 12 months. Biopsy collection was performed in ...

visualization of bacteria in bladder biopsy sections via fluorescence in situ hybridization

Source: Neugent, M. L. et al., <a href="https://app.jove.com/v/60458">Detection of Tissue-resident Bacteria in Bladder Biopsies by 16S rRNA Fluorescence In Situ Hybridization.</a>&#160;J. Vis. Exp. (2019)
This video demonstrates the visualization of bacteria in deparaffinized bladder tissue from a urinary tract-infected patient. Fluorescently-labeled probe hybridized with bacterial 16s ribosomal RNAs reveals the tissue-associated bacteria and their distribution, providing valuable spatial insights into their density within the sample.

Visualization of Bacteria in Bladder Biopsy Sections via Fluorescence In Situ Hybridization

All procedures involving sample collection have been performed in accordance with the institute&#39;s IRB guidelines.
1. Tissue Preparation for Fixation ...

Begin with a glass slide carrying a deparaffinized and rehydrated infected urinary bladder biopsy tissue section containing uropathogenic bacteria.
Introduce fluorescently-labeled universal probes &#8212; a complementary single-stranded oligonucleotide specific to the conserved region of bacterial 16s ribosomal RNA, or 16s-rRNA.
Incubate the slide in the dark at an elevated temperature for optimum hybridization.
During incubation, the probes exclusively hybridize to the conserved region on bacterial 16s-rRNA, imparting green fluorescence to the bacteria.&#160;
Post-hybridization, wash with a pre-warmed buffer to remove the unbound probes.
Stain the tissue section with Hoechst dye and fluorophore-labeled cellular protein markers.
Hoechst molecules bind to the DNA, staining the nuclei blue, while fluorophore-labeled markers interact with specific cell components such as mucin and actin filaments, imparting red fluorescence in the biopsy section.
Wash the slide and add mounting media to the tissue section.
Under a confocal microscope, green fluorescence within the red fluorescence-carrying bladder biopsy section confirms the tissue-associated bacteria.