Instruct: Identify the main category of Biorxiv papers based on the titles and abstracts
Query: A powerful and versatile new fixation protocol for immunohistology and in situ hybridization that preserves delicate tissues in planaria Whole-mount in situ hybridization (WISH) is a powerful and widely used technique to visualize the expression pattern of genes in different biological systems. Here we describe a new protocol for ISH and immunostaining in the planarian Schmidtea mediterranea. The new Nitric Acid/Formic Acid (NAFA) protocol is compatible with both assays and prevents degradation of the epidermis or blastema. Instead of proteinase K digestion, formic acid treatment is used to permeabilize tissues and preserve antigen epitopes. We show that the NAFA protocol successfully permits development of chromogenic and fluorescent signals in situ, while preserving the anatomy of the animal. Further, the immunostaining of different proteins was compatible with the NAFA protocol following fluorescent in situ hybridization. Finally, we demonstrate with high resolution confocal imaging that the regeneration blastema is preserved when using the new method. This new NAFA protocol will be a valuable technique to study the process of wounding response and regeneration.
5
The categories of this paper are: cell
17
The categories of this paper are: molecular
12
The categories of this paper are: behavior
10
The categories of this paper are: and
27
The categories of this paper are: pathology
23
The categories of this paper are: immunology
26
The categories of this paper are: biochemistry
9
The categories of this paper are: synthetic
13
The categories of this paper are: animal
32
The categories of this paper are: zoology
4
The categories of this paper are: neuroscience
1
The categories of this paper are: bioinformatics
6
The categories of this paper are: plant
21
The categories of this paper are: toxicology
18
The categories of this paper are: systems
11
The categories of this paper are: cognition