The credit of write up @Sailekshmi
I have copy and pasted the write up sent in CUBE What’s app group with the objective to archive the data and documentation and I also encourage @Sailekshmi to post the summery on metastudio.
CUBE ChatShaala Meeting Summary
Date : 11th June 2025
Key Discussion : Identification and screening of sleep mutant fruit flies using home lab setups and behavioral studies.
Main Presenter : Ayana (Nallalam, Kozhikode)
Collaborators Noted : Akanksha, others involved in whiteboard synthesis
Concepts Explored
Wild-Type vs Sleep Mutant Fruit Flies
Wild-Type Flies : Active during daytime, sleep at night.
Sleep Mutants : Show opposite activity — awake and active at night, rest during the day.
Key indicators : Mating, climbing, feeding observed at night for sleep mutants.
Experimental Tools
BRSV & TRSV Bottles : Used to maintain different cultures.
Banana peel in 500 ml bottles as attractant for wild types.
Tomato slices as bait in mutant screening ( Ayana’s setup ).
Ayana’s Home Lab Experiment
Objective : To trap and culture native sleep mutant fruit flies.
Setup Date : 7th June 2025, 10 PM
Bait : Tomato slice
Observations at Different Times:
Night : Almost no flies trapped till 3 AM.
Morning ( 8 AM–2 PM ): Sharp increase; 43 flies trapped by 1:30 PM.
At 3 AM, two gravid ( egg-carrying ) flies were successfully trapped.
Single Line Culture Results
Bottle A : 3 AM flies transferred → After 2 days → 30 larvae observed.
Bottle B : Mother fly transferred separately → No larvae observed → Suggests fly still active (not laid eggs yet).
This strongly supports the hypothesis that the flies trapped at 3 AM were gravid and were active only at night — indicating possible sleep mutant behavior.
Life Cycle Recap (Observed in A Bottle)
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Gravid fly → Egg (0.3 mm)
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1st Instar Larva (1 mm)
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2nd Instar (2 mm)
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3rd Instar (5 mm)
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Pre-Pupa → Pupa → Adult
Note : Larval stages are motile, while pupal stages are non-motile.
Akanksha’s Screening Proposal
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Set up screening of mutant flies at 3 AM (feeding, climbing, mating expected).
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Re-check activity at 9 AM.
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Concept of “Sleep Mutant” being probed by monitoring activity patterns across time points.
Queries for Reflection and Further ChatShaala Discussions
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How can we distinguish sleep mutants from early risers or late sleepers among wild types?
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Is a single time-point (e.g., 3 AM) enough to confirm sleep mutation? Or do we need a full circadian activity chart?
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Why did Bottle B show no larvae even after 3 days—what could have happened to the egg-laying process?
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Can temperature of Nallalam (Ayana’s place) influence sleep-wake cycles in native fruit flies?
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Would replicating the experiment with different baits (banana vs. tomato) influence results?
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How can Ayana’s experimental strategy be improved for scaling screening of more suspected mutants?
ChatShaala Summary – 11 June 2025 (For School Cubists)
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What Are “Sleep Mutant” Fruit Flies?
Normal fruit flies are awake during the day and sleep at night.
Sleep mutants do the opposite : they’re awake at night and sleep during the day.
- Ayana’s Home-Lab Setup
She used two plastic bottles (500 ml) with tomato slices as bait.
Bottle A & Bottle B were set at 10 PM on 7 June.
Aim : to catch flies that are awake at night and maybe laying eggs.
- What Did Ayana Observe?
Nighttime ( 11 PM–3 AM ): Almost no flies seen.
Early morning ( 8 AM–1 PM ): Many flies appeared—up to 43!
At 3 AM, two flies carrying eggs (called “ gravid ” flies) were caught—this is a big clue!
- What Happened Next in Bottle A & B?
Bottle A : After 2 days, found 30 baby fly larvae—meaning the flies laid eggs!
Bottle B : After 3 days, still no larvae—so the flies didn’t lay eggs there.
This suggests the flies caught at 3 AM were indeed sleep mutants, active at night.
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Life Cycle of a Fruit Fly (Observed)
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Egg (tiny, about 0.3 mm)
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1st-Stage Larva (1 mm)
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2nd-Stage Larva (2 mm)
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3rd-Stage Larva (5 mm)
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→ Pre-pupa, then Pupa, then Adult fly
- Akanksha’s Screening Plan
Check at 3 AM for feeding or egg-laying—that’s when sleep mutants should be active.
Check again at 9 AM, when normal flies are usually awake.
This helps us compare and confirm which flies are mutant.
Questions to Think About
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How can we be sure a fly is a sleep mutant and not just sleeping late?
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Should we check at more times (like midnight, 6 AM) to be confident?
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Why did Bottle B have no larvae—what might have happened?
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Could the temperature or home location (like Kozhikode) affect sleep patterns?
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What if we use a different bait (like banana) instead of tomato—would it change who comes?
