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

Published: April 30, 2023

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…

Disclosures

The authors have nothing to disclose.

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

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Cite This Article
Eosin Staining of Soft-Tissue Samples: An Eosin-Based Procedure for Whole Mouse Kidney Staining to Visualize Specific Tissue Microstructures. J. Vis. Exp. (Pending Publication), e21035, doi: (2023).

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