I am somewhat new to working with the Pioreactor and my next set of experiments will require working with some pretty strict anaerobes. We have considered some options and would appreciate some advice or ideas from people who have maybe worked with anaerobes in the Pioreactor. So info: we will be using the pumps for flow of media in and waste out, and will want to sample from the Pioreactor hourly (for qPCR experiments down the line). Here are our thoughts of how to set this up:
Option 1: to set up the Pioreactors in our anaerobic chamber. This would be good so as to not introduce oxygen into the system when sampling, BUT would reduce the number of Pioreactors we could have in our cluster as it is quite tight for space/ difficult to work in.
Option 2: to consider using Vial Cap B outside of the anaerobic chamber, but we are then not sure how we could go about sampling without opening the vial and introducing oxygen. One thought that crossed my mind was if there was a way to have Vial Cap B with a rubber, self-healing plug, which is see isn’t an option as of yet.
We would prefer to work outside the anerobic chamber, but any and all advice would be much appreciated
Thanks!
The vial cap B isn’t supported anymore (I can explain why). The current vial cap A and S are somewhat air tight, but probably you don’t want to risk it. You can try to add high-temp silicone in the obvious air gaps (near the steel tubes, and also where the luer lock connector attaches to the steel tube). Also: a newer design of the vial cap S has a better placed o-ring under the vial cap, which is much more airtight (I think we made this change in June).
We managed the sampling by installing a T derivation in the downstream of the outlet, after a non-return valve, closing the other end of the tube (which leads to the waste reservoir) with a Hoffman clamp. By this way, air cannot enter into the Pio. We take advantage of output pump to fill a sterile luer syringe connected to the T derivation.
Hi Cameron!
I’m taking advantage of this post to find out why vial B is no longer supported. It’s the one we have in our pioreactor and we really like it because of the safety —in terms of asepsis— that it offers.
“no longer supported” sounds harsher than I meant. I meant we are not manufacturing the current iteration of them¹. Four reasons:
they are harder to 3D print: they require printing upright, which means we need support structures on the underside. Sometimes this print fails.
Because of the design + print orientation, the barbs are easy to break off (printed along the barb axis => weakest point)
The tubes needed for the vial cap B aren’t what we use: we use 1/16" ID 1/8" OD tubing throughout the Pioreactor system, but these require larger OD tubing.
Liquid touches the plastic - this is probably okay, but we’d rather have liquid touch something more biocompatible: glass, metal, or silicone.
That being said, we are current reimagining what a new vial cap B would look like, keeping in mind the above.
Thank you for the clarification. My apologies for referring to the fact as “no longer supported.” I didn’t mean to sound harsh, I just wanted to know the reasons (which are logical, by the way).
