Apr 20, 2024

Public workspaceProtocol for Mouse IP Anesthesia (Ketamine-Xylazine) V.2

DOI
dx.doi.org/10.17504/protocols.io.14egnzr9qg5d/v2
  • Haowei Li1,
  • Markus Holzl1,
  • Mohsen Khosravi-Maharlooei1,
  • Nichole Danzl1,
  • Austin Chen1,
  • Megan Sykes1
  • 1Columbia Center for Translational Immunology, Columbia University, New York
Sandy Beshir
Open access
DOI: dx.doi.org/10.17504/protocols.io.14egnzr9qg5d/v2
Protocol Citation: Haowei Li, Markus Holzl, Mohsen Khosravi-Maharlooei, Nichole Danzl, Austin Chen, Megan Sykes 2024. Protocol for Mouse IP Anesthesia (Ketamine-Xylazine). protocols.io https://dx.doi.org/10.17504/protocols.io.14egnzr9qg5d/v2Version created by Sandy Beshir
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: April 20, 2024
Last Modified: April 20, 2024
Protocol Integer ID: 98525
Funders Acknowledgement:
NIH
Grant ID: U01 DK123559
Abstract
This protocol details the constitution of our anesthesia cocktail that we use for sedating our mice when performing surgeries. The surgeries we typically perform are thymectomies and implantation of material under the kidney capsule (through open abdomen or minimally invasive approaches).

Note
The changes of this protocol was made by Austin Chen on Markus Holzl original protocol.


Note
Corresponding Authors

Mohsen Khosravi-Maharlooei
Email: mkm2182@cumc.columbia.edu

Austin Chen
Email: ac4274@cumc.columbia.edu
Tel: 425-283-6900

Materials
Materials:

Anesthesia is a mixture of ketamine and xylazine in 1x PBS.
  • ·Ketamine: Ketathesia (100 mg/mL; cat: 056344; Henry Schein)
  • ·Xylazine: Xylazine sterile solution (20 mg/mL; Columbia ICM)
  • ·1x PBS: Phosphate Buffered Solution (1X) (cat: BW17516F24; Lonza)
  • ·0.22 um filter: Syringe Filter Unit, 0.22 µm, polyethersulfone (Millex-GP; cat: SLGPR33RS)


Preparation:
Ketamine stock: 100 mg/mL
Xylazine stock: 20 mg/mL

Desired ketamine concentration: 0.08 mg/gram of mouse
Desired xylazine concentration: 0.012 mg/gram of mouse
  • We will make anesthesia solution to dose Amount10 µL anesthesia/gram of mouse for IP injections

For 10 mL of anesthesia:
  1. Amount0.8 mL Ketamine (100 mg/mL) + Amount0.6 mL Xylazine (20 mg/mL) + Amount8.6 mL 1x PBS
  2. Filter solution with 0.22 um Millex-GP filter

Calculations:
  • Ketamine: 0.8 mL * 100 mg/mL in 10 mL → 8 mg/mL Ketamine → 0.08 mg/10 uL
  • Xylazine 0.6 mL * 20 mg/mL in 10 mL → 1.2 mg/mL Xylazine → 0.012 mg/10 uL




For Intraperitoneal (IP) Approach:
For Intraperitoneal (IP) Approach:
Use 10 uL/gram of anesthesia

Comments:
1. It takes time until the mice fall asleep

2. Very often, mice do not fall asleep but move around like zombies. This requires another small dose of IP ketamine-xylazine injection or a small dose of IV injection

a. When doing IP injection again, we have injected IP with a volume of about 1/3 of original volume

b. For IV approach after IP, it takes very little anesthesia for them to fall asleep. We usually use around 10-20 uL. Inject this slowly into the mouse. The mouse should fall asleep rapidly. It is very easy to overdose with IV injection so be careful.