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).

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