Eosin Staining of Soft-Tissue Samples: An Eosin-Based Procedure for Whole Mouse Kidney Staining to Visualize Specific Tissue Microstructures

Abstract

Source: Busse, M. et al., 3D Imaging of Soft-Tissue Samples using an X-ray Specific Staining Method and Nanoscopic Computed Tomography. J. Vis. Exp.  (2019)

This video demonstrates the procedure for the acidification of the whole mouse kidney with fixation followed by Eosin Y staining. This cytoplasm-specific Eosin staining method helps visualize and identify anatomical structures in the tissue.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. Eosin staining protocol To fixate soft-tissue samples, fill a 50-mL centrifuge tube with a fixative solution containing 9.5 mL of 4% (v/v) formaldehyde solution (FA) and 0.5 mL of glacial acetic acid (AA). NOTE: Prepare the FA solution freshly from a 37% acid free FA solution stabilized with appr…

Materials

50-ml centrifuge tube by Falcon	VWR	734-0453
Formaldehyde solution, 37%	Carl Roth	CP10.2	Acid-free, stabilized with ~10% MeOH
Glacial acetic acid	Alfa Aesar	36289.AP
Eosin Y disodium salt	Sigma-Aldrich	E4382	Certified by Biological Stain Commission
Phosphate Buffered Saline (PBS)	Merck	L1825	Dulbecco's formualtion, w/o calcium and magnesium
Sample Tubes by Nalgene	Carl Roth	ATK5.1
Rocking Shaker ST5	CAT	60281-0000
Cellulose tissue paper	VWR	115-0600
Microcentrifuge tubes by Eppendorf	VWR	211-2120	Safe-lock, 2.0 ml
Ethanol absolute by Baker Analyzed	VWR	80252500
Petri dish by Sterilin
Forceps, by USBECK Laborgeräte	VWR	232-0096
Plastic pasteur pipette	Carl Roth	EA68.1	Graduated, 1 ml