Lab support

Getting started

What you will receive to begin assays

  1. Standardized 3D printed platform to conduct assay
  2. Hemotek 1 mL blood reservoir (product code: R29P10)
  3. Hemotek blood-feeding membrane (product code: MEM5)
  4. Peltier heating base
  5. Test compounds and DEET (positive control) shipments, including safety data sheets. Please run DEET as a positive control to confirm that the assay is detecting repellent behavior.
  6. Standard template for recording data and access to shared repositories to upload your lab’s assay data
  7. Assay protocols, video training and support, and help with data formatting and analysis.

What you will need to provide

  1. Reliable access to a standardized wild-type Liverpool strain Aedes aegypti colony (if you are in the U.S., we can provide this to you). If your lab has access to another colony and would prefer to use that for assay data, please reach out to us to move forward.
  2. Temperature- and humidity-controlled behavioral testing room maintained at 28 ± 2°C and 50–70% relative humidity (RH).
  3. Please ensure that the behavior chamber has air purifiers or is connected to a fume hood/exhaust system.
  4. Insect holding/rearing cages
  5. Sheep or bovine blood. You will need approximately 1 mL per assay run: 6 mL per compound for the treated runs, plus 2 mL per assay day for the two shared controls.
  6. Mosquito netting or mesh (if applicable)
  7. Micropipettes (P20, P200, and P1000) and sterile pipette tips
  8. Glass vials or microcentrifuge tubes for solution preparation
  9. Timer
  10. Aspirator for insect transfer
  11. Thermometer and hygrometer
  12. iPhone 14 or newer or Logitech Brio 4K-HD webcam
  13. Camera mount
  14. Cages (BugDorm-4E1515 Small Insect Rearing Cage)
  15. Control solvent (1% ethanol in water)

Assay

Preparing the insect cages

  1. Use a modified BugDorm® Small Insect Rearing Cage as the assay cage. One mesh side of the cage shall be removed and replaced with a transparent acrylic panel to facilitate video recording and accurate observation of mosquito landings during the assay.
  2. Ensure that each cage is clean, dry, and free of residual odors or chemical contamination before use. Inspect the acrylic panel and mesh for damage and confirm that the cage is securely sealed.
  3. Label each cage with the assay identification number, treatment group, replicate number, date, and operator initials.
  4. Using an aspirator, transfer 20 healthy, host-seeking mated female mosquitoes of 1–3 weeks age into each cage. Minimize handling to reduce stress and avoid injury to the insects.
  5. Allow mosquitoes to acclimate to the assay room under standardized environmental conditions for 30–60 minutes before initiating the assay.

Preparing the mosquito landing surface

  1. Inspect the mZero-provided 3D-printed platform for structural integrity and a smooth, defect-free finish before use.
  2. Clean the landing platform before each assay and allow it to dry completely.
  3. The 3D-printed platorm is heat resistant up to 80°C, allowing it to be heated between assays to remove residual volatile compounds and minimize cross-contamination without damaging the arena.

Arena layout (top-down view of the platform)

Top-down diagram of the mZero assay arena showing its inside zone, band, red rim, and slotted surface.

Assay configurations

Side-by-side cross-sections of the contact and non-contact mZero assay configurations.

Side cross-section of the same 3D-printed platform in the two assay configurations: (A) contact form and (B) non-contact form.

Control assay procedures

  1. Fill a Hemotek blood-feeding reservoir with sheep or bovine blood and securely mount the membrane according to the manufacturer's instructions.
  2. Place the Hemotek reservoir on a heating pad or Hemotek heating unit and preheat the blood to 34–37°C before initiating the assay.
  3. Position the blood feeder beneath the pre-cut opening in the foam board so that the membrane containing the warmed blood is the only accessible heat source for the mosquitoes.
  4. Place the mosquito cage containing 20 mated, host-seeking female mosquitoes and the 3D-printed platform on top of the blood feeder, ensuring that the blood feeder is aligned directly beneath the central mesh opening of the landing arena. This arrangement allows mosquitoes to detect host-associated cues while preventing direct access to the blood meal.
  5. Run one contact and one non-contact control each day and use these controls for all chemicals tested that day.
  6. For the non-contact control, apply 50 µL of 1% ethanol in water to the inner surface beneath the landing zone.
  7. For the contact control, apply 50 µL of 1% ethanol in water directly to the landing surface.
  8. Allow the solvent to evaporate completely before starting each control assay.
  9. Once the cage is properly positioned, begin the assay immediately.
  10. Record mosquito behavior for 5 minutes using a top-mounted camera through the transparent acrylic panel of the modified BugDorm cage.

Contact and non-contact assay procedure

Non-contact assay

  1. Prepare the test chemical at 1% concentration using 1% ethanol in water as the solvent.
  2. Apply 50 µL of the diluted test chemical to the inner surface beneath the landing zone, ensuring that the treated surface is not accessible to the mosquitoes.
  3. Allow the solvent to evaporate completely before starting the assay.
  4. During the assay, mosquitoes are exposed only to the volatile (airborne) components of the test chemical and are unable to make physical contact with the treated surface.
  5. Position the mosquito cage on the blood feeder and begin the assay.
  6. Record mosquito behavior for 5 minutes using a top-mounted camera through the transparent acrylic panel of the modified BugDorm cage.

Contact assay

  1. Prepare the test chemical at 1% concentration using 1% ethanol in water as the solvent.
  2. Apply 50 µL of the diluted test chemical directly onto the 3D-printed platform within the designated landing zone.
  3. Allow the solvent to evaporate completely before starting the assay.
  4. During the assay, mosquitoes are able to land on and physically contact the treated surface, allowing them to detect both the volatile and non-volatile components of the test chemical.
  5. Position the mosquito cage on the blood feeder and begin the assay.
  6. Record mosquito behavior for 5 minutes using a top-mounted camera through the transparent acrylic panel of the modified BugDorm cage.

Watch assays

Control
Contact
Non-contact

Compounds

Monthly shipments

You will receive 10 compounds (not including DEET) to begin your assay work after your application and delivery details are approved. After data from the first 10 compounds has been recorded and checked by the mZero team, each lab will receive 20 compounds each month. Each set of compounds will include unique candidates our models rank as most likely to stop a mosquito from landing, including a shared panel of compounds sent to multiple labs to establish inter-lab variance.

Your lab is the importer of record for chemical shipments. Tell us in advance if it caps quantities or restricts any hazard class.

Handling & storage

  1. Store compounds as labeled on arrival; keep them in the dark and at the labeled temperature.
  2. Let vials equilibrate to room temperature before opening to avoid condensation.
  3. Prepare working dilutions fresh on the day of the assay. The standard screen uses 1% ethanol as solvent unless the protocol sheet says otherwise.

Safety

Every compound will be shipped with a safety data sheet including GHS classification. The shipment will be classified and packed to IATA/DOT standards. mZero will disclose structure to an EHS officer on request.

Handle all compounds with standard chemical-lab hygiene: gloves, lab coat, eye protection, and fume-hood preparation of dilutions.

Compound delivery details are submitted as part of the Request Access application.

Insectary

Testing each compound requires six treated runs: three non-contact and three contact, with 20 age-matched adult female mosquitoes per assay, for a total of 120 females per compound. Each assay day requires two additional shared control runs, one non-contact and one contact, using 40 females. Use these daily controls for all chemicals tested that day.

The insectary should maintain enough healthy, age-matched adults for the treated runs, the shared daily controls, routine colony maintenance, and failed runs. At an approximately 1:1 sex ratio, plan for at least 240 adults per compound plus at least 80 adults per assay day before accounting for mortality, failed runs, or colony maintenance.

Data

Sharing your data

  1. Your workspace is created when you register on the Lab Portal. The portal shows its Lab ID and the cloud storage structure <Lab ID>/Metadata/ and <Lab ID>/Video Footage/.
  2. Complete the metadata worksheet with your assay results. Download template here, or download the portal copy prefilled with your Lab ID in the first lab_id column. Name it [lab_id]_metadata_[date].xlsx; for example mzero4821_metadata_2026-08-13.xlsx. Submit one worksheet per assay day rather than appending to a running file.
  3. Choose the completed manifest in the Metadata slot. CSV, TSV, XLSX, and JSON files are read in your browser before transfer so the portal can check required values, the Lab ID, compound and batch IDs on treated runs, the two shared daily controls, duplicate filenames, and required columns. Correct and re-select a file as often as needed. Unusual but valid values, such as 31°C, are warnings and do not block submission. Submit remains unavailable until the manifest has no blocking errors and all eight matching videos are selected.
  4. Select all eight MP4, MOV, or WebM assay recordings together: six treated runs for the compound, plus the shared daily contact and non-contact controls. Use [lab_id]_[date]_[treatment]_[condition]_[replicate]; for example mzero4821_2026-08-13_DEET_contact_01.mov. Camera defaults like IMG_0028.mov are not accepted. Every selected filename must appear exactly once in the file_name column of your metadata; mismatches are blocked before transfer.
  5. Upload within 24 hours of the run completing or of a minimal quality review where one is needed. Submit positive, negative, and inconclusive results alike.
  6. Keep your source video files for 12 months so runs can be re-scored if the analysis pipeline is updated.

Accepted metadata formats in the portal: CSV, TSV, XLSX, and JSON. Accepted video formats in the portal: MP4, MOV, and WebM. Each successful transfer receives a traceable receipt with its folder, object name, generation, checksum, and validation status.

Data Quality

All labs run a fixed protocol on the same assay, use one contact and one non-contact solvent control for every chemical tested that day, and test a shared compound panel at the same concentrations.

We will report the following metrics to assess data reliability:

  • Inter-assay correlation (r)
  • Hit re-test rate
  • Inter-lab variance (CV)
  • z-score

Data analysis and compound ranking

Compounds will be ranked by a cumulative landing-zone occupancy metric. Each frame of the assay video footage shows the number of mosquitoes in the landing zone (inside plus red ring) multiplied by the frame interval. Summed across the assay, this gives cumulative mosquito-time in the zone, measured in mosquito-seconds: six mosquitoes present for one second is six mosquito-seconds. Each compound is scored as the percent reduction in occupancy against the matching daily contact or non-contact control.

Total mosquito-seconds = sum((n_inside + n_on_band) × dt).

The video footage also supports finer-grained behavioral measures, including spatial distribution, distance from the treated surface, entry and exit rates, dwell time, and how these change over the assay.

Accessing data

Metadata files and video footage from every participating lab are readable by all labs; you have edit access to your own lab's folder only.

Use the Lab Portal's recent uploads section: select this lab for the current workspace, all labs for research uploads shared across the registered network, and refresh after a new submission. The portal is the normal access path. A cloud storage console link works only for a Google account that has separately been granted direct bucket access.

Metadata worksheets and video footage stay within the network and are not redistributed publicly.

This is a cloud storage workspace managed through the mZero portal, not a Google Drive folder.

Commons

Repellent compound identities will be published under CC BY 4.0. Metadata files and video footage are shared across all participating labs.