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…
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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 |
