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Updated: Jul 31, 2025

Use of a Low-flow Digital Anesthesia System for Mice and Rats
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Mouse isoflurane anesthesia using the drop method.

Maya J Bodnar1, Anna S Ratuski1, Daniel M Weary1

  • 1UBC Animal Welfare Program, Faculty of Land and Food Systems, University of British Columbia, Canada.

Laboratory Animals
|May 5, 2023
PubMed
Summary
This summary is machine-generated.

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The drop method effectively anesthetizes mice using lower isoflurane concentrations (1.7%–3.7%). This method provides anesthesia faster at higher concentrations, but further research is needed to assess mouse aversion.

Area of Science:

  • Veterinary Anesthesiology
  • Laboratory Animal Science

Background:

  • Isoflurane anesthesia is recommended for mouse euthanasia via carbon dioxide, but vaporizer access can be challenging.
  • The 'drop' method offers an alternative by introducing a fixed volume of isoflurane into an induction chamber.
  • Previous studies indicated 5% isoflurane via the drop method is effective but aversive; lower concentrations remain untested.

Purpose of the Study:

  • To evaluate the efficacy and behavioral effects of lower isoflurane concentrations (1.7%, 2.7%, 3.7%) using the drop method for mouse anesthesia.
  • To determine the time to achieve surgical plane anesthesia at different isoflurane concentrations.
  • To assess stress-related behaviors during induction with the drop method.

Main Methods:

  • Male Crl:CD-1 (ICR) mice (n=27) were randomly assigned to receive isoflurane via the drop method at 1.7%, 2.7%, or 3.7%.
Keywords:
Euthanasiaanestheticinductionrefinementrodent

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  • Measures of anesthetic depth (recumbency, righting reflex, pedal withdrawal reflex) and stress behaviors (rearing) were recorded during induction.
  • Statistical analysis using Least Squares Means was performed to compare induction times between groups.

    • Main Results:

    • All tested isoflurane concentrations (1.7%, 2.7%, 3.7%) effectively induced anesthesia in mice.
    • Increasing isoflurane concentration significantly reduced the time to recumbency, loss of righting reflex, and loss of pedal withdrawal reflex.
    • Rearing, a stress-related behavior, was most pronounced immediately after isoflurane administration across all groups.

    Conclusions:

    • The drop method is effective for anesthetizing mice with isoflurane concentrations as low as 1.7%.
    • Higher concentrations (up to 3.7%) expedite anesthetic induction.
    • Further investigation is warranted to evaluate potential aversion associated with the drop method at various isoflurane concentrations.