# Protocol for whole-mount immunostaining of brains of the eastern subterranean termite, Reticulitermes flavipes

## TL;DR

This paper provides a detailed protocol for immunostaining termite brains, enabling visualization of protein patterns using confocal microscopy.

## Contribution

The study introduces a new protocol for whole-mount immunostaining specifically tailored for Reticulitermes flavipes termite brains.

## Key findings

- The protocol includes steps for dissection, fixation, staining, and mounting of termite brains.
- The method produces immunostained brains suitable for confocal microscopy.
- The approach can potentially be adapted for other insect species.

## Abstract

Immunostaining is a powerful technique for visualizing tissue morphology and protein expression patterns, but in non-model organisms, it may be impeded by a lack of established protocols. Here, we present a protocol for whole-mount immunostaining of termite brains that we applied to the termite Reticulitermes flavipes. We describe steps for brain dissection, fixation, staining, and mounting. This approach generates stained termite brains that can be visualized through confocal microscopy and can potentially be adjusted to suit other insect species.

For complete details on the use and execution of this protocol, please refer to Merchant and Zhou.1

•Whole-brain immunostaining protocol for use in Reticulitermes termites•Steps for dissecting, fixing, staining, and mounting termite brains•Produces immunostained brains that can be visualized via confocal microscopy

Whole-brain immunostaining protocol for use in Reticulitermes termites

Steps for dissecting, fixing, staining, and mounting termite brains

Produces immunostained brains that can be visualized via confocal microscopy

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

Immunostaining is a powerful technique for visualizing tissue morphology and protein expression patterns, but in non-model organisms, it may be impeded by a lack of established protocols. Here, we present a protocol for whole-mount immunostaining of termite brains that we applied to the termite Reticulitermes flavipes. We describe steps for brain dissection, fixation, staining, and mounting. This approach generates stained termite brains that can be visualized through confocal microscopy and can potentially be adjusted to suit other insect species.