Vibratome Sectioning of Whole Murine Kidney: A Technique to Generate Thin Sections of Murine Kidney

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

1. 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.
   NOTE: KRB can be stored at 4 °C for up to one week and should be pre-bubbled with a mix of 97% O₂ and 3% CO

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
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