ºù«Ӱҵ

General information

• Fly Lab D22: 18-19°C (0114 22) 22811
• Fly Growth Media Room: D34a  32802
• Culture Rooms: D22b 18°C, 70%RH; D22c 25°C, 70%RH

Fly facility technicians

• Kath Whitley (Fly Facility Manager) k.whitley@sheffield.ac.uk
• General fly technician email flytechnicians-group@sheffield.ac.uk

Other useful contacts

• Martin Zeidler m.zeidler@sheffield.ac.uk (fly academic lead)
• Melanie Hannah m.hannah@sheffield.ac.uk (School Technical Manager)

The risk assessments and CoSHH forms for the fly lab and media room are in labelled blue files in the relevant rooms.

General fly lab rules

If the CO₂ sensor is sounding then this indicates dangerous levels of CO_2. Leave the lab immediately and prop open the door. 

Please ensure you turn off your CO₂ if you are not using it and after you have finished using your fly station.

No food or drink is to be consumed in the laboratory.

When listening to music via headphones please keep the volume down so that others cannot hear it and to ensure you can still hear any alarms that may sound.

Please swab down your bench with 70% IMS after you have finished your fly work each day.

When you have finished using your microscope make sure the light source is switched off.

If you find any mites please tell Kath so the problem can be dealt with immediately.

Only keep fly cultures for the maximum time indicated below. This will be checked weekly by fly technical staff and you will be informed if items are close to the maximum times.  Technical staff will contact you rather than disposing of any items that have single copies or are part of stock collections.  If there is an emergency please communicate this so we can make other arrangements to flip unique fly stocks or move items to your research lab.  Any remaining items will be disposed of by the date given.

Fly cultures can be kept for a maximum of:

• all bottles and vials  at 25°C - 3 weeks
• straight sided bottles at 18/19°C - new bungs 5 weeks; old bungs 4 weeks
• ICRF bottles at 18/19°C - all bungs 5 weeks
• vials at 18/19°C - 6 weeks

18°C controlled temperature room – for main stock collections only unless for specific experiments on a short term basis, agreed in advance by Kath Whitley or Cherry O’Keefe (in Kath’s absence).

Trays to be labelled with the researcher's name.  Trays of vials or bottles tipped together can be dated on the front of the tray or the first vial in the row.  Individual bottles and all vials bundled together by elastic bands need to be dated individually.

Please put all used trays next to the food on the long bench for checking and washing by fly technicians.  Put all small ties in the labelled beaker so they can be reused. 

Please wash up any egg laying pots and re-usable yeast containers.

Empty your GM waste bin at least every 2 weeks and label the lid with the date you have done this.

If the fly food in the main lab is running low it is your responsibility to get more from the cold room D33.

Ensure you fully close controlled temperature room doors (the light should not flash).

Keep a record of your fly food use (either individually or on whole lab record).

Media and equipment

Fly growth media: 

Please ensure you have ordered all fly food on the shared spreadsheet at least a week before you need to use it.  Please use the earliest dated (ie oldest) food first. 

If you require large amounts of media for an experiment please coordinate with Kath or Cherry at least two weeks before you wish to use it.  Fly media is stored in the cold room D33.  If you need to use small quantities of foam stoppered vials (FST), some will be available in D22 (on the long bench next to the door).  If this is running out or you need larger quantities please fetch more from the cold room.  Cotton bunged vials (CST) will be left in the cold room.  Please take what you have ordered and leave the rest in the cold room for others to use.

Fly stock collections

Please refer to the General Fly Lab Rules section for details about the maximum time fly cultures can be kept for and correct labelling.

18°C controlled temperature room – for main fly stock collections only unless for specific experiments on a short term basis, agreed in advance by Kath Whitley or Cherry O’Keefe (in Kath’s absence).

Remember much of the stock collection is irreplaceable. It is very important to avoid contaminating or otherwise harming stocks.

Please always get permission from the relevant Principal Investigator before taking stocks – this may also ensure you get the stock you really need!

Do not deplete the flies from a stock, to the extent that it threatens the health of the stock. In general, it is OK to take flies from the back vial, provided the other vials are healthy (you should check this!). Only take flies from other vials with great care.

Do not remove stocks from the collection, except briefly when collecting flies and always return promptly. Be very sure to return stock to the correct position in the tray.

Any flies that are obtained from sources outside the laboratory must be quarantined before bringing them into the fly lab to make sure they are not carrying anything that can spread to other stocks. Allow them to go through 2 generations and examine the culture carefully under a microscope to check for mites before bringing them into the fly lab or culture rooms.  Mites can out compete weak fly stocks or even kill them so please be vigilant and tell Kath if you see old cultures that need to be dealt with or if you discover any mites.  Then we can stop them spreading.

Adult mites are smaller than fly embryos and often walk on the sides of vials and bottles around the pupae and empty pupal cases.  Mite eggs and adults can be observed in Drosophila culture using the youtube link below.

If you know your cultures contain mites and you don’t need them, freeze the vials overnight to kill the mites.  Place the frozen cultures into a yellow waste bin.  Also freeze any fly cultures from outside sources before disposing of them.

If your culture contains mites and you wish to keep the flies, remove cultures from the fly lab. Passage them three times on consecutive days onto fresh medium.  Then allow the flies to go through two generations.  Putting the tubes on a surface of glycerol makes the transfer of mites from one tube to another more difficult.

Waste

Please refer to the poster by the door and waste assessment in D22 for information regarding the correct segregation and disposal of biological waste.

Please collect vials and bottles with the new style foam bungs in a tray after use and put them onto the trolley between the controlled temperature rooms.  We can then freeze, wash and reuse the bungs.

Use the GM waste bins to dispose of other fly cultures.  These will contain an orange bag on the outside and two black bags on the inside.  Please empty your GM waste bin at least every 2 weeks and label the lid with the date you have done this.  To do this, take out the bags and twist the top around.  Secure very tightly with a long electrical cable tie ensuring no flies can escape.  Fill out an orange label (GM/Infectious Waste) with the following information: Department-BIS Laboratory–D22 and Date Sealed.  Place the label on the bag and put it out of the way to the right of the incubator.   

Put an orange bag and then two black bags into the yellow bin.  The waste bags are collected from the fly lab on Monday, Wednesday and Fridays.

The sharps bin is located on the long bench.

Use the bin with black lid for non-hazardous waste and the bin with green lid for recycling.

Rinse used Fisher Winchesters and dispose of the lid.  Deface the label with a cross and place the Winchester outside the lab for collection.

Carbon dioxide

If the carbon dioxide alarm is sounding:

Do not enter the fly lab.  If you are in the fly lab when the carbon dioxide alarm is sounding exit the lab immediately.  Wedge door open to ventilate the lab.

In emergency internal x 4444, external/mobile 0114 2224444

If the alarm is flashing but not sounding the CO₂ will have exceeded the 8 hour safe long term exposure limit (0.5%).  Do not work in the lab for extended periods under these conditions.  Do not wedge the door open as this breaches GM regulations.

Please remember to turn off your CO₂ tap when you have finished using it.

CO₂ supplies can be turned off to each bench in the fly lab by closing the black valves underneath the benches.

Culture room alarms (D22b and D22c)

The temperatures of D22, D22b and D22c are monitored.  Alarm notifications are sent to technical staff so we will be aware of them.  If you are working in the lab, you can mute these locally using the mute button on the control panel between D22b and D22c.

The alarm will go off if the temperature or humidity goes above or below limits or there is a power failure.  If there is a temperature fault we may open the culture room door until the fault is rectified. 

If the autodialler rings the fly lab phone nearest to the door, report this to Kath.  Once this has been reported press 8 on the phone to cancel the message.  If you do not get to the phone in time please enter the default code 1234 on the alarm panel to stop it phoning around everyone.

D22 Air Conditioning

The fly lab temperature should read 18°C on the thermometers above Kath Whitley and David Strutt’s work stations.  If the temperature on the thermometers reads below 18°C or above 19°C inform Kath Whitley or Cherry O’Keefe immediately.  We will then adjust the air conditioning settings.