Cleaning the SmartSPIM Imaging Chamber
Imaging chambers need to be cleaned between refreshes of the imaging medium and or when switching to a different imaging medium. The recommended cleaning schedule is once per month for EasyIndex and once every 3 months for EasyIndex Matched Immersion Oil.
Protocol
1. Remove the objective from the chamber. Wipe off the imaging medium (EasyIndex, Immersion Oil, etc.) with a lens cleaning wipe. Wet a new cleaning wipe with 100% methanol or similar cleaning alcohol and gently wipe the lens so there are no remaining streaks of imaging medium. Set the objective aside; you may place the cap on it to avoid dust accumulation.
2. Remove the imaging chamber from the microscope. Carefully pour the contents into a waste disposal container.
3. Wash the imaging chamber with warm soapy water. A couple drops of regular dish soap is fine. You can use your gloved hand to gently rub the glass to remove anything sticky. Do not use a washcloth or sponge, as they could scratch the glass or leave particles behind.
4. Rinse the chamber with tap water, then with distilled water.
5. You may turn the chamber over and leave it to air dry for half an hour, or you may dry out the inside with a lens cleaning wipe or similar non-scratch, residue-free towel. Avoid using anything that could leave behind paper particles or dust inside the chamber.
6. Once dry, use methanol and lens cleaning wipes to wipe out the inside of the chamber, removing any water spots that you can see. Take special care with the glass windows (if you have a machined metal chamber) to remove all spots, streaks, and dust. Wherever the laser passes through the glass, take special care to make sure it is extremely clean so the laser will pass through the glass without interference. Hold the chamber up to the light to help you see any particles or streaks.
7. When you are satisfied that the glass is clean and dry, you may refill the imaging chamber with your preferred imaging medium and slide it back into the microscope.
8. If your imaging medium is EasyIndex, pour a layer of mineral oil over the EasyIndex (less than a quarter inch thickness is plenty) to prevent EasyIndex evaporation. You may need to wait for a few hours for the bubbles to settle out of the EasyIndex before you can start imaging again.
[Tip] If there is a bubble stuck under the objective, bend a plastic zip tie and use it to sweep the bubbles out from under the objective. Be gentle!
9. Check that the objective is clean, then remount the objective and insert it into the chamber. Make sure your imaging medium is high enough to cover the surface of the lens.