You have the answer...mesh sandwich and it will cost you close to nothing.

i'd slice it ASAP and then keep them cold (0-5C) and well oxygenated and also consider increasing the glucose concentration

We've been doing that to keep cortical tissue alive but we were closer from the extraction site, so we did not have to worry too much about transportation :). However, we still had a chamber connected to a small bottle of carbogene to carry in good conditions the tissue. The tissue we had was thicker than yours and we processed the slicing just after the tissue arrived in the lab, and we were able to keep them all over the night in oxygenated ice cold acsf. You may ask Andrew K. Tryba for more details about the transportation chamber we used (see my contacts).

I would really go for a mesh sandwich and be careful to have no bubble going too much under the tissue for it not to be floating too much. In addition, you may know that you can saturate the air and the liquid in O2 even if you do not inject the carbogene in the liquid directly. Some kind of chamber may fit a system in which you would still have the aCSF oxygenated, without having to deal to much with these bubbles (I do that now to keep some slices from mouse in my new projects).

We did not used glucose or sucrose enriched media and we had some nice human cortical neurons (see http://www.ncbi.nlm.nih.gov/pubmed/21076335).

Use 0-5C cold ACSF with sucrose replacing Na. Also you may add ketamine to block NMDA channels. In such solution and temperature oxygenation is less critical. So you can transport thick tissue without slicing to 300 um. After transportation you need to slice the tissue immediately(in the same Na free ACSF) and then keep it at 34c for 30 min. In the same solution. This will help to boost up ATP in the cells. Then transfer to normal ACSF and the tissue slice can stay at 26-28c, viable for many hours.

Just a suggestion for the mesh. Women's nylon tights work well for supporting slices. They can be cut into areas of whatever shapes you need, e.g. stretched over a perspex annulus to support the slices. It is crucial to wash the nylon (and any perspex) in water for a day or two to wash out any toxins. If the mesh is too small, ladder the tights.

Experiment with having the slices lying on a mesh soaked in the solution you choose, and irrigate from below, while keeping the air/gas above saturated with water.

You can use gibbs chamber.

Some years ago Per Magnelind has made magneto- (SQUID) and electrophysiological measurements on hyppocampal slices at Chalmers University Göteborg, Sweden. In his thesis (Chalmers University 2006) he has written something about the tissue slice transportation. Please find attached a copy of this chapter. Unfortunately I do not have a digital version of his thesis.

Hi,

I used to do it years ago, having to drive from the North of Paris to the South (45 min).

Slices were cut in the hospital (we were carrying the slicer).

If you want to oxygen it, the best solution is to use an interface holding chamber, more than easy to build with a 3D printer (I can send you a picture), just add some ACSF at the bottom and bubble it to keep the slices moist (they won't move on the filter).

However, I know that Richard Miles used to carry the whole block of tissue in pre-oxygenated ACSF from the Hospital to the lab (30 min by metro). This led to the classic Cohen et al, Science paper. Hence, it seems that you don't need anything funky. I guess that as long as you maintain the block in solution saturated with O2 it can survive quite a while.

Hi Stuart,

We have transported numerous human neocortex samples between the operating theatre and our lab. The methods have been published (see Teichgräber et al. 2009). Essentially, we use a modified ACSF for transport, carbogenated and temperatures near 5 degrees C. For small samples we use Falcon 50 ml tubes filled up to the rim closed tightly.

Time for transport used to be about 20 min (change in temperature from 4 to 6 degrees). Now operations take place at another, more distant site and tranport time is between 30 and 40 min. Both, sharp microelectrode recordings and patch clamping works fine. The tissue is typically viable for about 24 hrs, sometimes stable neurones can be obtained long passed the 24 hrs.

Good luck

Rudolf

Thank you all for your insight,it really has been most helpful.

Best,

Stuart