☑️S12 Assembly and Sample Prep

How to put together your Omni-Stainer™ S12 system and prepare samples for staining

Step 1 - Create Humid Environment: add 30 mL deionized water to the Omni-Stainer™ S12 base

This helps to maintain a humid environment and prevents evaporation

Step 2 - Construct S-type Flow Cell: mounting a Cover Pad on a slide with a sample.

Adhesive spacers on the plastic Cover Pads ensure optimal flow cell thickness. The chosen adhesive allows the Cover Pad to slide when wet but holds it in place to prevent lateral staining solution bleed.

To place the coverpad

First remove the coverpad from the plastic film. Then, pre-wet both the coverpad and the slide surface. Then slide on the coverpad over the sample such that the "flow cell" is evenly filled with the liquid (PBS or hydration buffer) without any bubbles (see videos below).

There are multiple methods to build the flow cell "sandwich," aiming to ensure the Cover Pad sits firmly on the slide, and the internal "capillary chamber" contains buffer without any air bubbles.

The "Yury Method"

https://vimeo.com/843291880vimeo.com

Yury Method, slightly more finesse than the "Easy Method" but fits into tissue prep workflows and saves labware.

The "Easy Method"

https://vimeo.com/751961225vimeo.com

Easy Method, *Note: Cover Pads now have peelable plastic film on both sides, rather than peel tabs.

Easy Method" consists of sliding on the Cover Pad under the layer of PBS or any such compatible buffer inside a suitable reservoir such as a pipette tip box lid.

The "Big Drop Method"

As confidence and experience are built, constructing an S-type flow cell can also be achieved by placing a big drop (250ul) of compatible buffer on top of the slide and then sliding the coverpad over the drop to avoid bubbles.

If bubbles are entrapped – sliding the coverpad such that the bubble is released and then sliding it back after adding a bit more of the liquid allows to "expel" the bubbles

Preparing different sample types

• FFPE sections: a sample needs to be baked, deparaffinized with Xylenes or a suitable alternative, and re-hydrated. If HIER is needed, at this time we recommend it to be. It is possible to do HIER directly in Parhelia Omni-stainer using the Thermal Sheath, but that functionality is experimental and hasn’t been fully tested. Please reach out to us directly at [email protected] if you would like to try running HIER with Parhelia Omni-stainer.

• Fresh frozen sections: sample needs to be fixed and re-hydrated

• Cell culture/cell spreads: samples needs to be fixed and re-hydrated

Step 3 - Place flow cell into the shelf, and insert shelf into base

Place flow cell into shelf

Place shelf into base

Step 4 - Adhere moisture barrier paper to the lid's bottom, wet it with 10 mL of deionized water

Make sure the holes are aligned if using automation lid

Humidity barrier placement for manual lid

Step 5 - Put the lid on the base, matching slanted corners

Align the slanted and rounded corners of the lid and base.

Full Assembly

Insert sample, then put on lid (automation lid shown)

S12 Automation Specs

S12 Manual Specs

Using a Thermal Sheath?

Thermal Sheath & Temperature-controlled incubations

Done! You're now ready to choose Manual or Automated mode and launch the demo!