Volume of Clearing Solution

Andy_Donaldson

How much clearing solution are you all using for the ETC step? And how long are you performing the ETC step? The procedure says "several days," but have any of you determined an optimum time? Thanks!

nicolasrenier

My setup uses 5.5L of solution. 4L are necessary to will the water bath, about 1L goes in the filter tank, and the rest in the chamber and pipes. I haven't figured out a good running time, and it seems a much discussed issue for everyone so far. I depends on the size and nature of the sample, on the temperature and voltage of the run... In my experience, 4 days to 1 week is a good place to start at 30V and 37°c...

aysegul

Hi, nicolasrenier. How big is your filter tank and what is the volume of your water bath ? Is 5.5 L enough volume for the ETC? Thanks!

Fabio

My tank has 5.5L volume and ist enough. Normally I use just 5L. I think the water bath from the publication has a volume between 5-7,5 L so its enough.

aysegul

Thank you Fabio. What is your water bath brand name?

Fabio

The one from the paper ist the Lauda-Brinkmann RE415, I use a model that has heating and no cooling function. This is the Lauda-Brinkmann A6

aysegul

Fabio, i guess cooling is not neccessary at 37 °c. What is your conditions for clearing
(voltage, temperature, time) ?

Fabio

Until now im still optimizing, but the starting pint is 20 V, 37 °C, min 6 days

Fabio

btw these parameters depend on your ETC chamber, until know im using an exact copy of the one described in the publication

aysegul

thank you so Fabio