X-Message-Number: 5077 From: Date: Tue, 31 Oct 1995 11:45:30 -0500 Subject: hyperbaric freezing Thanks to Wowk and Darwin for comments on mildly hyperbaric freezing. Continuing: First, as to Darwin's question about pulling a vacuum after cooling a specimen to near zero centigrade; he asks if that will cause liquid to freeze instantly at a higher temperature. No; reducing pressure from one atmosphere doesn't affect the freezing point appreciably, only the boiling point. This is freeze drying, or near-freeze drying. Of course, evaporation is a cooling process, so pulling a vacuum will tend to produce freezing by this cooling process (high school teachers like to demonstrate simultaneous boiling and freezing), but you will not by any means get instant freezing or freezing at a higher temperature. Brian's point about the heat of fusion is one I remembered only after the spur-of-the-moment post, and presumably the process wouldn't work (on large specimens) for that reason unless the temperature before release of pressure is below (roughly) negative 80 C, since the heat of fusion of water is about 80 cal/gm. If we have to go that low, we will be dealing with pretty high pressures, and it is no longer so easy and cheap. However, it should still be much easier and cheaper than the temperatures and pressures used by Fahy, Waitz and others. Darwin says he gets around 2,000 atmospheres with a chamber large enough for a brain, the chamber with accessories costing $75,000. Greg Fahy has previously expressed the opinion that even a full-body capacity chamber at 2,000--5,000 atmospheres would be affordable in the context of cryostasis, after considering probable usage rates etc. Darwin says the small intracellular ice crystals formed by release of overpressure will rather rapidly coalesce into larger crystals (because of the dependence of vapor pressure on crystal size and geometry). How quickly? At - 80 C the vapor pressure of ice is pretty low (again, I don't have my references at hand) and hence the recrystallization should be pretty slow, one would think. Mike says he has watched the recrystallization under the microscope (at what temperature?), which suggests a matter of minutes or hours, but maybe not. To avoid the crystal growth, presumably the cooling to below around - 135 C (where the vapor pressure becomes negligible and essentially independent of crystal size) would have to be rapid relative to the rate of growth. This might be feasible. Further, one suspects that, since the tissues are mostly solid at this point, migration of water molecules would have to be pretty slow and there would be relatively little random churning, avoidance of which was the main idea in the first place. Yes, small intracellular ice crystals can do a lot of damage, the apparent damage being exaggerated by thawing effects and perhaps by artifacts of preparation, staining etc; but once again, the idea was not so much to minimize damage as to minimize degradation of information. Also, if we shift the damage somewhat from intercellular to intracellular, this may save relevant information, because the intracellular information is (I think) mostly generic and not unique to the individual, whereas intercellular and intertissue connections and structures in the brain are thought (at least by some) to represent our individuality and memories. Robert Ettinger Rate This Message: http://www.cryonet.org/cgi-bin/rate.cgi?msg=5077