tweeking the recipe and hours of cooking

protocolhell

Here is a shot of the new recipe/protocol in process. (Above). The solutions used for the immunohistochemical staining process after the mess has been cleaned up. (Below).

antibodies

Today was a full day and night in the ‘kitchen’ cooking up the pig skin with virus.
This time I finally figured out how to mix the antibody solutions properly thanks to Stuart (my collaborator for Living Viral Tattoos and  a research scientist based in the Anatomy and Human Biology department). Besides getting the skin stamped, transplanting the viral host cells and incubating them (aka cooking them in the oven), I was measuring and mixing solutions for hours. The non-stop pipetting action and washing the skin with PBS is mindnumbing work.

The kitchen metaphor is fitting for this process. Last week the human skin  puffed out after it had been in the incubator for four hours, losing its stamp shape. Megan, a  SymbioticA regular, saw the  bloated skin pages and commented that they were overflowing souffles.  I’m trying to created more of a ‘pancake’ so that the stamp designs hold in the viral host cells as long as possible.

Here is a short video clip of ‘cooking’ the pig skin with HaCat cells, Lentivirus and primary antobodies in an incubator.

transfectedThis photo is not in focus but here you can see how the skin looks after incubation. The stamp imprint is visible apart from the rest of the skin due to a reaction between the viral Hacat cells and the pig skin’s epidermal cells.  If the immohistochemical staining process works, this area will turn blue in the shape of the stamp design.

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