Mar 15, 2024

Public workspacePreparation for OCT embedding Mouse Kidney Tissue (TDA-University of Washington)

DOI
dx.doi.org/10.17504/protocols.io.6qpvr31b2vmk/v1
  • 1University of Washington
Runze Dong
Open access
DOI: dx.doi.org/10.17504/protocols.io.6qpvr31b2vmk/v1
Protocol Citation: kangsk, Runze Dong, Liangcai Gu 2024. Preparation for OCT embedding Mouse Kidney Tissue (TDA-University of Washington). protocols.io https://dx.doi.org/10.17504/protocols.io.6qpvr31b2vmk/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: March 14, 2024
Last Modified: March 15, 2024
Protocol Integer ID: 96734
Abstract
Preparation for OCT embedding Mouse Kidney Tissue (TDA-University of Washington)
Kidney tissue collection
Kidney tissue collection
C57BL/6J male mice from the National Institute of Aging Rodent Resource were handled according to the guidelines of the Institutional Animal Care Committee at the University of Washington.  Mice aged 5 and 26 months were selected for the study. Kidney tissue extraction was conducted immediately following euthanasia.
Make an incision then cut the skin from the bottom to the top of the mice followed the abdominal line with the round side of the scissors.
Proceed to cut the lateral skin to get access to the kidney. Carefully extract the kidney, rinse it with HBSS (Hank's Balanced Salt Solution, Thermo, 14025092), remove the capsule, and section the kidney into pieces measuring 3 to 5 mm in thickness.
OCT Embedding
OCT Embedding
 Apply a thin layer of OCT compound (Sakura Tissue-Tek O.C.T. Compound, Sigma, 4583) into a disposable mold (Epredia, 18985-1).
Position the sample tissue at the center of the mold, ensuring that the side intended for cutting faces inward.
Cover the tissue completely with additional OCT compound.
Lower the mold into liquid nitrogen, being careful to not fully submerge the mold. Hold in liquid nitrogen for a few seconds until fully frozen.
Transfer the frozen mold to dry ice for storage at -80ºC.