Teton and Teton Atlas™ workflows require users to assemble the flow cell prior to beginning a run. Two critical aspects users should take into consideration are completely drying the flow cell slide prior to flow cell assembly and minimizing the amount of time between drying the flow cell and starting the cytoprofiling run. Using slides that are not completely dry prior to flow cell assembly increases the risk of bubble formation on the slide. Imaged bubbles can lead to a reduction in data. Flow cells that are dry for >10 minutes may begin to see a reduction in protein counts. The longer the time a dry flow cell sits without starting the run, the greater the protein count reduction may be.
To facilitate the recommendation to load Teton flow cells within 10 minutes of drying, the following run setup workflow should be implemented. Start with a fixed slide and thawed Teton or Teton Atlas cartridge and reagents. Slides should be kept in PBS following fixation, and, if needed, they can be stored at 4°C for up to 30 days when stored with RiboLock RNase inhibitor. After removing the PBS, apply Teton reagents A and B per the instructions in the Teton CytoProfiling User Guide, leaving PBS from the last wash in the slide kit wells. Next, set up the run on the instrument and begin the prime. After priming has completed, dry the slide, disassemble the slide kit, and assemble the flow cell. Finally, start the run. (Image 1.)
Image 1. Teton and Teton Atlas run setup workflow.
When drying the slide prior to flow cell assembly, use a vacuum aspirator with a 200 µl tip to completely remove the PBS from the slide. Position the tip in the corner of the well along the wall and slide the pipet tip down the side of the frame until you reach the gasket. This may require aspirating multiple times, aiming for the same corner each time to avoid aspirating the cells. Consider using a multichannel aspirator if applicable.
Video 1. Aspirating liquid from the Teton Slide Kit
After drying the slide as much as possible, disassemble the slide kit by removing the slide clips, frame, and gasket. See Disassembling the Teton Slide Kit using the Teton Slide Kit Tool for a demonstration. It is possible to use the aspirator to remove any additional liquid remaining on the slide after disassembling the slide kit.
Video 2. Aspirating liquid from the slide after disassembling the Teton Slide Kit
If the user does not feel confident that they can aspirate liquid from the slide without touching the cells, they should proceed with flow cell assembly without additional liquid removal. An example of a dry slide is shown in Image 2. Assemble the flow cell as shown in the Guidance for assembling a flow cell using the Teton™ Flow Cell Aligner and Sealer knowledge base article and start the run within 10 minutes of drying the slide.
Image 2. Example of dry slide.