A powerful and versatile new fixation protocol for immunostaining and in situ hybridization that preserves delicate tissues.

ABSTRACT:

Background

Understanding how genes function to heal wounds and restore lost tissue is essential for studying regeneration. Whole-mount in situ hybridization (WISH) is a powerful and widely used technique to visualize the expression patterns of genes in different biological systems. Yet, existing methods to permeabilize samples for WISH can damage or destroy fragile regenerating tissues, thereby preventing such experiments.

Results

Here, we describe a new protocol for in situ hybridization (ISH) and immunostaining in the highly regenerative planarian Schmidtea mediterranea. This new Nitric Acid/Formic Acid (NAFA) protocol is compatible with both the assays and prevents degradation of the epidermis and regeneration blastema. The NAFA protocol achieves this without the use of p

SUBMITTER: Guerrero-Hernandez C 

PROVIDER: S-EPMC11533299 | biostudies-literature | 2024 Nov

REPOSITORIES: biostudies-literature