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 /…
Offenlegungen
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
Vibratome Sectioning of Whole Murine Kidney: A Technique to Generate Thin Sections of Murine Kidney. J. Vis. Exp. (Pending Publication), e20550, doi: (2023).