Flinders Microscopy Room 4E421 FMIAF Optical MicroscopySOP002 22 Feb 2007 J. Clarke

/ Safe Work Method Statement
FLUORESCENCE MICROSCOPES
Olympus BX50, AX70 and IX71 Fluorescence Microscopes in Room 4E421
(Use in association with Risk Assessment FMIAF Optical Microscopy RA002)
S.O.P NUMBER / RISK ASSESSMENT NUMBER / RISK ASSESSMENT
FMIAF Optical Microscopy SOP002 / FMIAF Optical Microscopy RA002 / Medium
DATE CREATED / REVIEWED / Employees Involved in SOP Creation / CONTACT PERSON

15.07.13

/

Jennifer Clarke

/

Yvette DeGraff

Personal Protective Equipment Required
None. Samples or chemicals requiring PPE are not to be brought into the Fluorescence Microscope facility
Before Work Commences
Users musthave received authorisedtrained and the training documented
(Training is available through Flinders Microscopy Unit )
You must have read and understood this SOP, the associated Risk Assessment (MicroscopyRA002) and the usernotes for this equipment.
Your samples must comply with the “Sample Preparation” guidelines as outlined in the usernotes for this equipment.
(Copies of all relevant documents are located by each fluorescence microscope in Room 4E421)
Hazards
Postural damage Risk Assessment MEDIUM
Eye damage from UV light Risk Assessment MEDIUM
Mercury vapour exposure Risk Assessment LOW
Cuts from broken glass (slides / coverslips) Risk Assessment LOW
Risk of damage to equipment
Risk of damage to objective lenses. Prevention: Lower stage (upright microscopes) or lower objective turret (inverted microscopes) before changing specimens. Only use immersion fluid with correct objective types.
Safe Procedures
TO USE
  • You must comply with the “before work commences” criteria on the previous page
  • Follow instructions below for booking the equipment, use of the mercury lamp, use of the microscope and additional SOP requirements as described below
  • Do not attempt to modify any of the equipment. Maintenance to the equipment is only to be performed by authorised personnel (Flinders Microscopy staff and Biomedical Engineering).
1. BOOKINGS AND CANCELATIONS:
You must follow the rules for booking of the fluorescence microscopes to ensure correct use of the mercury lamp. Specifically;
  • You must book your session.
  • If you are not going to be using the mercury lamp, you need to specify this in your booking as “Not using mercury lamp”.
  • If you can’t make it to a session that you have booked, it is your responsibility to ensure your session is cancelled (to prevent the Mercury Lamp being unnecessarily left on).
  • If you cancel “at the last minute”, it is also your responsibility to ensure the Mercury Lamp isn’t unnecessarily left on; check it hasn’t been left on for you when you cancel your session
  • The maximum time you may bookthe equipment for is two hours. After two hours of work at the microscope/computer you are required to take a break (see 4. below). If the equipment is available you may continue to use it providing you have taken a break.
2. USE OF THE MERCURY LAMP
  • Before turning on, check that the accumulated time of use is less than 800 hours (according to the Mercury Lamp Time Sheet). If the accumulated time exceeds 800 hours do not use equipment but notify Flinders Microscopy staff who will replace the lamp.
  • Before turning on, check that the lamp has been off for at least 30 minutes (according to the Mercury Lamp Time Sheet). If it has been turned off within the last 30 minutes you must wait until it has been off for 30 minutes before turning it on.
  • Upon turning the Mercury Lamp on, write the time turned on (to the nearest 5 minutes) and your initials on the Mercury Lamp Time Sheet.
  • Before turning the Mercury Lamp off, check the booking calendar to see if someone is using it after you. If someone is using it within up to 1 hour of you finishing, leave it on. Otherwise, turn it off so long as it has been on for at least 30 minutes (see below).
  • Do not turn the Mercury Lamp off within 30 minutes of being switched on. If you finish your work within 30 minutes (and no-one is booked to use it in the next hour), you must wait until it is safe to turn it off.
  • Upon turning the Mercury Lamp off, write the time off (to the nearest 5 minutes), your initials and the accumulated lamp usage time the Mercury Lamp Time Sheet.
  • If in the unlikely event a Mercury Lamp explodes (you will hear a very loud “bang”) turn the Mercury Lamp off and evacuate the room immediately, closing the door behind you. Place the “no entry” sign across the door and notify Jen Clarke, Ian Gibbins or Kerry Gascoigne immediately.
3. USE OF THE MICROSCOPE
Note: This document does not constitute a manual or training guide. Users of this equipment must receive authorised training and use this Safe Operating procedure in conjunction with the usernotes for this equipment and the risk assessment for this equipment.
  • Turn on the microscope itself (if required)
  • Keep the shutter closed when the mercury lamp light source is not required
  • When the shutter is open, never look directly into the light coming through the objective lens, ie, never look directly into the light path
  • With the shutter closed and the microscope stage lowered (upright microscopes) or the objective turret lowered (inverted microscopes), put your sample on the microscope stage.
  • With either brightfield or fluorescence light (shutteropen), adjust the stage/objective height to bring your specimen into focus, by viewing through the eyepieces. Take particular care not push the objective lens onto the slide beyond the focal point.
  • Close the shutter and lower the stage (upright microscopes) or the objective turret (inverted microscopes) before changingthe specimen on the microscope stage. (This is also therefore how the microscope should be left when you have finished).
  • On the Olympus IX71 inverted microscope you must also lower the objective turret in between changing objective lenses, and take particular care that the objective lenses do not hit the metal plate on the stage
  • Acquire images as desired using the camera and computer
4. ADDITIONAL SOP REQUIREMENTS FOR USE
  • Maintain correct posture whilst working at the microscope and computer (refer to )
  • Do not work at the microscope/computer for more than 2 hours without taking a break
  • Only use immersion oil if necessary. Immersion oil must only be used with “oil immersion lenses”, do not use with dry objective lenses. Use only non-toxic Olympus immersion oil and remove excess from objective lens and wash hands after use.
  • Fixed specimens must be mounted with a coverslip, and be sealed, clean and dry. This means that slides must be free from excess mounting media and that nail varnish must have had at least 60 minutes to set.
  • Live cells may be viewed on the Olympus IX71 inverted fluorescence microscope (but not on the upright Olympus BX50 or AX70 fluorescence microscopes. Live cells must be transported correctly. Where applicable, OGTR regulations must be complied with and the specimens suitably contained.
  • No chemicals are to be brought into the fluorescence microscope facility. This is not a wet lab area and materials which require personal protective equipment (eg gloves, safety glasses) should not be brought into this area.
  • Microscope slides are to be carried in an appropriate container, and broken slides or coverslips are to be disposed of immediately in the sharps containers provided.

END OF SOP

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