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emission 520 nm) was read in the upper and lower chambers, and the permeability coefficient was determined using the following formula: 22 P dextran =( RFU lower / RFU upper )( V )(1/ t )(1/ A ), where RFU is the relative fluorescent units in the upper and lower wells, V is the volume of the bottom well, t is the time that the FITC-dextran was allowed to diffuse, and A is the total surface area of the monolayer (cm 2 )

Swabbing with alcohol before every single puncture is not optional safety theater

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Once you puncture the rubber stopper with a needle for the first time, the clock starts