🥇Sample Mounting Instructions

To ensure bubble-free operation, use these directions for S-type and C-type flow cell construction / sample mounting.

*This information is also in S12 Assembly and Sample Prep and C12 Assembly and Sample Prep.

Sample Mounting for S12 Parhelia Omni-Stainer™

Two adhesive spacers running along the edges of the single-use plastic Cover Pads are calibrated to maintain the optimal thickness of the flow cell. The adhesive is carefully chosen in such a way that when wet the Cover Pad can slide along the surface of the slide and yet when slightly pushed against the slide the adhesive holds the position of the coverpad and prevents the lateral bleed of the staining solutions.

How to construct S-type Flow Cell: mounting a Cover Pad on a slide with a sample.

Placing a Cover Pad

First remove the Cover Pad 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 "Simple Method"

"Simple 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 "Yury Method"

The Yury Method can be used with classic or glass Cover Pads, but is especially useful if using Glass Cover Pads which have lower tack.

S-type flow cell preparation for Parhelia Omni-Stainer™ S12 Module Yury Method uses slightly more finesse than the "Simple Method" but fits into some tissue prep workflows more easily and is especially useful if using glass Cover Pads.

Plastic "Classic" Cover Pad (deprecated) methods

Just in case anyone has these out there. We only sell the glass now, since the plastic can generate gas bubbles in heating.

As confidence and experience are built, constructing an S-type flow cell can also be achieved by placing a big drop (250-500ul) 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. It is also possible to do HIER directly in Parhelia Omni-stainer using the Thermal Sheath and temperature module.

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

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

Additional Flow Cell Prep Videos

Yury Method with Classic Cover Pad [Video Link]

Big Drop Method with Glass Cover Pad [Video Link]

Yury Method, as explained by Yury - Glass Cover Pad

How to construct C-type Flow Cell: mounting a cover tile on a Support Pad | C12 Parhelia Omni-Stainer™

https://vimeo.com/751961194vimeo.com