Microscopes

A note from Anna:

"The number of users for the microscopes in lab has increased quite a bit. To ensure efficiency, I have created two booking calendars: for the katyperryscope and the nissanmicrascope. If you think of better puns we can rename them. 

You will get access to these calendars once you’ve been ‘trained’ on the relevant microscope. 

See image above. The one one in SEB606 (left) is katyperryscope, the other one (right) is nissanmicrascope. The nissanmicrascope has the micromanipulators on it and is currently downstairs"

Rules: 

• Be considerate

• Be organised

• Be courteous

• Be reasonable

• If you need it for a time-lapse or other long acquisition, book time overnight or start early in the morning to impact fewer users that might want to use it during the day time.

• If you need it for longer than 10 mins, please BOOK TIME

• If you need it for longer than 1 hour, please BOOK TIME at least 1 day in advance

• If you need it for less than 10 mins, check the calendar before bringing your sample into lab 

• If you change your your booking time, please update the calendar and alert other users

Protocol:

• BOOK the microscope as per rules above

• Dispose of chemistry gloves in the bin underneath the bench. 

• Wash your hands before using the microscopes with ungloved hands OR use clean gloves (make sure they are dust-free!).

• Do not use immersion oil unless you are using the 100x objectives. Use sparingly and please clean with specialised lens wipes before you leave. NEVER use kimwipes never ever.

• Turn everything off once you’re finished. Checklist:

  • White LED

  • Fluorescence LED

  • XY/joystick stage controller

  • Camera

  • Heating stage (if using)

• When imaging: 

  • Be gentle! Do not mash the objectives into glass, the objectives are expensive and can break.

  • Do not move the position of the condenser knob unless you realign it (and if you don't know what these words mean - don't move it).

• Leave the area cleaner than when you arrived.

  • Throw your samples out

  • Close pipette tip boxes

• You may need to get pipettors from next door and/or tips from the far end of 605 (our lab benches are the ones closest to Hilmer). If so please return the pipettors themselves back to 605. The tips can stay in 606.

• For data acquisition, saving temporarily onto a local drive is recommended for faster read/write speeds. For longer term storage, please store your data on the Z drive. This is backed up regularly and can be accessed off-campus. For instructions on the group share, please see the New Researcher Checklist.

For cleaning the objective (or any optics):

Refer to this guide: https://www.youtube.com/watch?v=-RetyUAWGc0

Waste disposal:

• Dispose of glass slides in the sharps bin

• Dispose of pipette tips etc in the solid waste bin

• Dispose of other hazardous waste somewhere you know how to deal with them (maybe that’s your own lab, or in 605)

If you encounter any problems or issues contact Anna. If you have any suggestions please also contact Anna.

Some important notes from JBM 2024:

Firstly, everyone using the scope should understand Kohler alignment. (see: https://www.olympus-lifescience.com/en/discovery/how-to-align-khler-illumination-in-6-simple-steps/) ("8. Center the Condenser" on page 22 of the manual)

Secondly, everyone using the scope for phase contrast images should understand how that alignment works, and how to troubleshoot issues. (see: https://www.microscopyu.com/tutorials/phase-contrast-microscope-alignment). (page 23 of the manual)

A recent issue was that multiple people have reported issues with phase contrast on the 100x. We have diagnosed the issue to be due, in all likelihood, to two separate issues:

Firstly, the Condenser Aperture Diaphragm (page 14 of the manual) above the phase contrast rings was closed. When doing phase contrast microscopy it must be fully open. (mentioned on pages 23, 24, 43, 44, etc of the manual)

Secondly, there was a second 100x objective on the Katyperryscope. The second one had come from the Nissanmicrascope and does not list a Ph value on it; it is not capable of phase contrast imaging.

The scope is, as far as we can tell, in perfectly working order at this stage. When you have a chance, please familiarise yourselves with the components of the microscope and how to troubleshoot any common issues you may have. Remember that people may use the scope for different sorts of imaging, so things such as a 1.5x tube lens and the condenser aperture diaphragm are good things to check when you hop on.