Assessment of Chemical-Induced Colitis via Immunohistochemical Analysis of Colon Tissue Sections

Abstract

Source: Du, Y. W. et al., Effects of Taste Signaling Protein Abolishment on Gut Inflammation in an Inflammatory Bowel Disease Mouse Model. J. Vis. Exp. (2018)

This video demonstrates the assessment of chemical-induced colitis via immunohistochemical staining of mouse colon tissue. In a stained tissue section, the presence of a large number of immune-responsive cells in the damaged tissue represents the infiltration of immune cells during colitis.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.

1. Preparation of Mice and DSS

1. Keep the knockout (α-gustducin^-/-) mice and age-, gender-, and body weight-matched wild-type control (α-gustducin^+/+) C57BL/6 mice individually in clean cages.
   NOTE: The knockout mice have been backcrossed with C57BL/6 mice for over 20 generations and have a nearly 100% C57BL/6 genetic background.
2. Dissolve 30 g of dextran sulfate sodium (DSS) powder in 1 L of autoclaved wate....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Antibody
CD45	BD Biosciences	550539
CD3	BD Biosciences	555273
B220	BD Biosciences	550286
CD11b	BD Biosciences	550282
Ly6G	BD Biosciences	551459
Reagent
Dextran Sulfate Sodium Salt (DSS)	MP Biomedicals	2160110
Streptavidin-HRP complex	BD Pharmingen	551011
Phosphate-Buffered Saline (PBS)	Sangon Biotech	B548117
BD 10 ml Syringe	BD Biosciences	309604
Instruments and equipment
Balance
Scissors
Forceps
Staining jars
Software
Imag-Pro Plus	Media Cybernetics, Inc.