💙"Blue Dye Demo" Automated Protocol
This demo protocol demonstrates the basics of automated capillary gap staining liquid exchange with an C12/S12 Omni-Stainer™
Last updated
This demo protocol demonstrates the basics of automated capillary gap staining liquid exchange with an C12/S12 Omni-Stainer™
Last updated
If you did not upload the custom labware to Opentrons, you will see a protocol analysis error.
The labware definitions needed for the demo are below for your convienance. The labware may need to be imported one at a time. Check out the Importing labware definitions guide for more info!
Remember to add 20-30 ml of distilled water to the Omni-Stainer base.
Make sure all labware (plates, reservoirs, Omni-Stainers) is flat on the liquid handler deck to stop tips from bending. SBS labware should fully click into or sit in the deck slot. Avoid usual mistake of back corners not fully in or level.
note: The videos above show violet dye in well 1 and clear in well 2, while this Blue Dye Demo protocol has clear (PBS) in well 1 and blue dye in well 2.
The Opentrons Deck Calibration Video has another good demonstration of jogging the pipette tip in the XYZ directions into the center of the engraved "+" symbol on the Opentrons deck is very similar to the labware calibration for the Omni-Stainer™ which is performed using the embossed "+" symbol on the C12 or S12 Omni-Stainer™ lid.
Ensure that the blue dye filling and washing is complete
Very small pockets of carryover buffer towards the bottom corners of the staining chamber are normal and should not affect staining quality because usually there’s no tissue present there. However, if you wish for a more complete exchange, you may wish to increase the wash volume as necessary.
If the flow seems abnormally slow, and the exchange seems incomplete and uneven:
Carefully check for bubbles in the chamber. Bubbles may form if the buffers used have significant amounts of dissolved gas. If bubbles are present, slide the sample up and down to release the bubble. If that doesn’t help, re-mount the sample. When in doubt, degas buffers before the run.
Make sure that the sample is positioned correctly within the staining chamber.
S12 Omni-Stainer: ensure the gray aluminum shelf is correctly positioned in the staining chamber, if this is off the liquid may be applied to the outside of the cover tile rather than into the capillary gap.
C12 Omni-Stainer: make sure that the coverslip rests evenly on both spacers, as in this diagram:
