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this is a video showing the use of the pad pen it is similar to a felt-tip marker only this draws a hydrophobic layer of material around your sample which concentrates the fluid overtop of your sample when we work with aqueous solutions even though water has a good surface tension it still tends to spread out and this dilutes the concentration of any expensive antibody or stain that you may be working with so what weamp;#39;re going to do is use a pad pen this pen pen contains 13 milliliters of a hydrophobic liquid that when drawn on the slide forms a barrier to hold aqueous solutions in a single spot when we first get it we need to prime the pen and weamp;#39;ll notice that by when the felt tip turns nice and red at this point we can draw around our sample at circle in this case or if we have a high aspect ratio sample we can draw it an oval we can draw a square you can draw whatever you want whatever your sample size shape might be so Iamp;#39;ve drawn