Log anesthesia events for a Drosophila cohort and classify the cumulative CO₂ exposure against the Bartholomew (2015) thresholds. Recommends recovery period before behavioral assays.
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Load example drosophila co2 anesthesia tracker data to see the full workflow
When to use
Do not use for
Even short exposures impair climbing and flight for up to a day. The single most common confound in fly behavioral data is unaccounted CO₂ exposure within the previous 24 hours.
Doing virgin collection and sex sorting in two separate sessions doubles the anesthesia events. Combine them when possible — collect virgins, sort by sex, set up the cross all in one CO₂ session.
CO₂ disrupts sleep architecture for at least 24 hours. Sleep researchers should use ice + chilled sorting plates exclusively.
Add a "CO₂" column to your fly stock notebook. Each entry should include date, duration, and purpose. Lets you audit the cumulative load on every cohort retrospectively.
Cumulative exposure is the sum of all event durations. Classification follows Bartholomew et al. (2015): cumulative <5 min → none, 5–30 min → transient (recover in 24 h), >30 min → persistent (recover in 48 h). The behavioral assay flag tightens warnings.
Last validated 2026-04-06. Calculations are designed for planning and documentation support; verify procurement decisions against manufacturer specifications or institutional SOPs.
ConductScience Drosophila CO₂ Anesthesia Exposure Tracker (v0.95.0). ConductScience, Inc. 2026. Available at: https://conductscience.com/tools/drosophila-co2-anesthesia-tracker
Bartholomew NR, Burdett JM, VandenBrooks JM, Quinlan MC, Call GB. Impaired climbing and flight behaviour in Drosophila melanogaster following carbon dioxide anaesthesia. Sci Rep. 2015;5:15298.
CO₂ anesthesia is the workhorse of fly genetics. It is fast (knockdown in seconds), cheap (a tank lasts months), and reversible (recovery in 1–5 minutes). But it is not without consequences.
CO₂ acts on the central nervous system through hypercapnic acidosis — the dissolved CO₂ lowers tissue pH, which inhibits synaptic transmission. Recovery requires the CO₂ to diffuse out and the buffering system to restore baseline pH. The process is reversible but not instantaneous.
Climbing performance was significantly impaired for ~24 hours after a 30-min exposure, and remained measurably reduced for some flies even at 48 hours. Flight performance recovered faster but still showed transient deficits. The conclusion: any CO₂ exposure within 24 hours of a behavioral assay is a potential confound.
Schedule any handling that requires CO₂ at least 24 hours before behavioral assays. For high-stakes assays (sleep, aggression), use cold anesthesia or unanesthetized handling instead. Document every CO₂ exposure in your lab notebook so you can audit the cumulative load on every cohort.
Place flies in a vial on ice for 5 minutes. They become immobilized and can be sorted on a chilled aluminum block. The recovery is slower (~10 min) and the throughput is lower than CO₂, but there is no metabolic disruption. The standard for behavioral cohorts.
A volatile anesthetic that knocks flies down for ~20 min. Less common in research labs because the recovery is slower and the long-term effects are not well characterized.
No anesthesia at all. Slower than CO₂ for sex sorting but the only way to maintain a fully naive cohort. Standard for circadian and sleep work.
Specialized chilled plates (e.g., FlySorter, Genesee Scientific cold pads) keep flies asleep for the entire sort duration. Higher throughput than ice-only and no chemical confound.
Tracks anesthesia history per fly stock and flags any cohort that has had CO₂ exposure within the recovery window before a scheduled behavioral assay.
Open app EquipmentCO₂ anesthesia pads, sorting plates, and chilled fly handling stations
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