# Protocol for the visualization of bacteria in the tick gut using whole-mount fluorescence in situ hybridization

## TL;DR

This paper provides a detailed protocol for visualizing bacteria in tick guts using fluorescence in situ hybridization.

## Contribution

A new protocol for whole-mount fluorescence in situ hybridization in tick gut tissues is introduced.

## Key findings

- The protocol includes dissection, fixation, hybridization, and imaging steps for tick gut visualization.
- The method can be adapted for use with other tissue types beyond the gut.
- Double-labeled probes (DOPE-FISH) are used for tissue hybridization.

## Abstract

The tick gut harbors a relatively diverse microbial community that includes commensal, beneficial, and pathogenic bacterial species. Here, we present a protocol for the visualization of bacteria in the tick gut using whole-mount fluorescence in situ hybridization. We describe steps for tick dissection, tissue fixation, hybridization, washing, and mounting. We then detail procedures for microscopy imaging of the whole-mounted samples. Although the procedure is designed for gut tissue samples, it can be readily modified for use with other tissue types.

For complete details on the use and execution of this protocol, please refer to Hodžić et al.1

•Steps for dissecting ticks and preserving tick tissues•Guidelines for tissue hybridization using double-labeled probes (DOPE-FISH)•Instructions for washing and mounting tissue samples•Procedures for microscopy imaging of whole-mounted samples

Steps for dissecting ticks and preserving tick tissues

Guidelines for tissue hybridization using double-labeled probes (DOPE-FISH)

Instructions for washing and mounting tissue samples

Procedures for microscopy imaging of whole-mounted samples

Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

The tick gut harbors a relatively diverse microbial community that includes commensal, beneficial, and pathogenic bacterial species. Here, we present a protocol for the visualization of bacteria in the tick gut using whole-mount fluorescence in situ hybridization. We describe steps for tick dissection, tissue fixation, hybridization, washing, and mounting. We then detail procedures for microscopy imaging of the whole-mounted samples. Although the procedure is designed for gut tissue samples, it can be readily modified for use with other tissue types.