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Welcome to the Novus Visual Protocol Series.
In this video you will learn how to perform all phases of florescent
immunocytochemistry using the most common methods for this assay.
Before starting the ICC procedure,
we will demonstrate how to prepare coverslips and cell culture plates
followed by the plating of our cells.
We begin by acid treating new coverslips in one molar of hydrochloride acid for
24 hours.
After carefully decanting off the acid and removing the glass coverslips,
we wash each with distilled water,
then a final rinse with ethanol.
As an optional step you can place the coverslips into a subbing rack submerge
in a .1 mg/ml solution gelatin or polylysine
for five minutes.
These coatings aide in providing additional adherence of cells to the
coverslips ensuring that they are not detached during later wash steps.
After treatment remove the subbing rack and dry the coverslips in the culture
hood or a heated oven.
Clean dry coverslips are then inserted into each well of a six well culture
plate
and covered with a lid.
To save time, large volumes of plates can be prepared ahead of time for later use.
The plates can also be sterilized by placing them under the hood's UV light.
With clean coverslips prepared in six well plates, we can now add our cells.
Platted here in cells add a density of one half million cells per well.
Cells are then cultured overnight in the incubator or until they reach your preferred
density.