🧑🔬Manual Operation and "Blue Dye Demo" Manual Protocol
The Manual Product Bundles and the Automated Product Bundles can both be operated manually with a P200 micropipette.
Follow steps 1-3 Assembly and Sample Prep guide for your Omni-Stainer type:
☑️S12 Assembly and Sample Prep☑️C12 Assembly and Sample PrepRemember to add 20-30 ml of distilled water to the Omni-Stainer base.
Now, exchange the solutions in the flow cell. (like videos)
To exchange the current solution in the flow cell with a new solution – pipette the new solution onto the slide or coverslip support pad surface slightly (~2mm) above the interface of the flow cell with air.
The new solution should be applied with a speed slow enough that it does not overflow over the surface of the coverslip or Support Pad. Ideally, the end of the pipette tip should be positioned close enough to the surface such that the solution immediately transitions from the tip to the surface avoiding the formation of drops.
Sometimes especially before sufficient proficiency in handling the Omni-Stainers is acquired slight occasional overflow over the surface of the flow cell may occur. This does not constitute any danger and does not impact the final staining quality
The laminarity of the solution flow in the flow cell starts as solution spreading across the upper edge of the Cover Pad or coverslip. The faster the solution flows the less is the diffusion at the exchange front and hence the less is the volume that is needed for complete solution exchange. Currently, the volume needed to replace the solution under the S-type Cover Pad is 150 μl and inside the C-type Support Pad is 100 μl.
Our engaging virtual demonstration serves as an excellent introductory guide for understanding the steps in operating the Parhelia Bio Omni-Stainer C12 and S12, encompassing both manual and automated usage, along with thermal sheath and thermal modules. View the virtual demo here!
Manual Blue Dye Demo
This is a very simple demo protocol that helps you understand how capillary gap exchange works inside Parhelia Omni-stainer and calibrate volumes that are needed for the gap exchange.
Manual Blue Dye Protocol
Prepare two [Eppendorf] tubes and fill one with PBS and the other with Blue Dye (1.0 mL each).
Utilizing a P200 pipette, gradually dispense 110 μl of Blue Dye onto the upper curved edge of the S12 Cover Pads (for C12 Support Pads, dispense 60ul of the solution onto the ramp). Observe attentively as the Blue Dye is drawn into the chamber, ensuring complete displacement of the clear liquid previously present in the staining chamber, which should drip down the wicking pillar. Subsequently, cleanse the staining chamber by dispensing 150 μl of the clear buffer, ensuring complete displacement of the blue dye.
Refer to the provided videos for visual guidance.
It is normal to have very small pockets of carryover buffer in the lower corners of the staining chamber, which should not affect the quality of staining since there is usually no tissue present in those areas.
However, if you desire a more thorough exchange, you can increase the wash volume as needed. If the flow appears unusually slow and the exchange seems incomplete and uneven:
Check for bubbles in the chamber. If bubbles are present, slide the sample up and down to release the bubble. If that does not resolve the issue, remount the sample.
Verify that the sample is correctly positioned within the staining chamber.
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