Vibratome Sectioning of Whole Murine Kidney: A Technique to Generate Thin Sections of Murine Kidney
Published: April 30, 2023
Abstract
Source: Grainger, N. et al. Isolating and Imaging Live, Intact Pacemaker Regions of Mouse Renal Pelvis by Vibratome Sectioning. J. Vis. Exp. (2021)
In this video, we demonstrate the vibratome sectioning of a whole mouse kidney to obtain thin slices. These sliced tissue sections can be stored at low temperatures until further downstream analysis.
Protocol
All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. Prepare Solutions Prepare 1 L of Krebs-Ringer Bicarbonate (KRB) solution containing 120.35 mM NaCl, 5.9 mM KCl, 15.5 mM NaHCO3, 1.2 mM Na2HPO4,1.2 mM MgCl2, 11.5 mM glucose, and 2.5 mM CaCl2. On the day of use, maintain the KRB solution on ice.<br /…
Divulgaciones
The authors have nothing to disclose.
Materials
| 24-well culture plate | ThermoFisher Scientific | 142485 | To store kidney slices in |
| 48-well culture plate | ThermoFisher Scientific | 152640 | To store kidney slices in |
| Absorbent paper | Fisher Scientific | 06-666A | To dry the kidney before applying glue |
| Cyanoacrylate glue | Amazon | B001PILFVY | For adhering the kidney to the specimen plate |
| B6;129S-Gt(ROSA)26Sor/J | The Jackson Laboratory | 13148 | GCaMP3 Mice |
| B6;129S-Gt(ROSA)26Sor/J | The Jackson Laboratory | 24105 | GCaMP6f Mice |
| B6.FVB-Tg(Myh11-cre/ERT2)1Soff/J | The Jackson Laboratory | 19079 | smMHC-CRE Mice |
| C57BL/6-Tg(Pdgfra-cre)1Clc/J | The Jackson Laboratory | 13148 | PDGFRa-CRE Mice |
| Fine-tip forceps | Fine Science Tools | 11254-20 | Used for fine dissection of kidney |
| Gillette Silver Blue double-edge blades | Amazon | B009XHQGYO | For insertion into blade holder of vibratome |
| Student Adson Forceps | Fine Science Tools | 91106-12 | For gently holding and moving the kidney |
| Student Dumont Forceps | Fine Science Tools | 91150-20 | Used for internal dissecting forceps |
| Vibrocheck | Leica | 14048142075 | Optional component for calibrating blade movement during cutting |
| VT1200 S Vibrating Blade Microtome | Leica | 14912000001 | Configuration 1 is used in our protocol |
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Vibratome Sectioning of Whole Murine Kidney: A Technique to Generate Thin Sections of Murine Kidney. J. Vis. Exp. (Pending Publication), e20550, doi: (2023).