In my last post, I concluded that switching the o-ring position from below the membrane to the top in the sepcon produced no leaks. The reason for this switch was to reduce the 1mm “dead-space” (created by the o-ring) that adds to the unstirred water layer (UWL). The next thing that Barrett and I looked…
In these trials, I used TER to look at the resistance between membranes with different amounts of pinholes. Finding a trend would produce a quicker way to scan for pinholes as opposed to checking each slot on the wafer with the microscope. The table below is my raw data. Wafer Number of Pinholes Resistance (ohms)…
The question arose last week about the affects of transport with use of the shaker in the Tecan, which led to a discussion about the current construction of the sepcon. The sepcon layout currently is as follows: The o-ring is 1 mm in diameter. Is this difference between the membrane and the receiver fluid dead…
Last week, I ran a few experiments to see how certain membrane conditions affected the transport of fluoroscein. The first experiment presented here contained three sepcons: one with 3 pinholes, one with only 1 intact slit, and another without slits to use for the background. The controls were HBSS and fluoroscein. The sepcon with 1…
Here is the excitation and emission wavelengths for two common fluorophores for transport: rhodamine and fluoroscein. rhodamine fluoroscein max ABS 480 nm 470 nm emission 558 nm 516 nm excitation 524 nm 492 nm The protocol for finding excitation and emission wavelengths of a fluorophore with use of the Tecan can be found here. During…
Before starting my transport project with the use of the Tecan, it was important to see how the Tecan worked with the sepcon and transwell kept in the well plate. The goals from this experiment were to: 1. Test for fluorescence from the plastic. 2. See if fluorescence from the apical side has an affect…