> CI Research Report - Independent Laboratory Research Findings

July-August 2000

The Cryonics Institute, over the last year, has initiated a new series of experiments intended to improve our cryopreservation procedures. We ship our specimens to independent laboratories for evaluation. We have one report in hand, and several others are expected momentarily, with many additional ones down the road.

In this month's issue of The Immortalist we publish just a portion of that report, with some comments. The full report will be on our web site as soon as we can get to it.

The first specimen was sent to the testing lab to see how they would perform, and the one chosen was out of curiosity-it used one of the methoxylated compounds reported by 21st Century Medicine as a potential break-through in vitrification. ("Vitrified"or "glassy" specimens are "solid" but without actual ice crystals of noticeable size, with a potential for reduced damage).

I emphasize that the procedure used in this preliminary test was not that used by 21CM. It was a simple perfusion of a sheep head with 20% 2-methoxyethanol, after a washout out with Ringer's. The head was frozen, not vitrified.

With these caveats firmly in mind, following are some brief excerpts from the report, including a couple of the micrograms (or micrographs if you prefer). Everything below was written or photographed by the testing lab professionals, not by CI personnel.

All specimens had been stored in liquid nitrogen at CI, then shipped in dry ice to the testing lab in Canada.

Again, the following is very brief and very preliminary, but something is better than nothing.

General. The inner-most and outer-most tissue appeared to have the greatest damage, while the intermediate zones had the best structure in terms of intact organelles and membranes.

Set 1. The tissue samples were fixed in glutaraldehyde and processed for TEM [Transmission Electron Microscopy], lead-stained.

Plate 26. Nuclei are generally intact. Cell membranes are entirely disrupted. Loose cytosol is everywhere. Mitochondria are evident, but other membrane bound organelles are not evident.

Set 2. Plate 15. High power view showing intact nuclear membrane and well organized chromatin structure even at this deep level. Cell membranes are clearly disrupted and the cytosol has been spilled. No membrane bound organelles are observed. Some intact tubules/filaments are evident passing by the nucleus.

Plate 18. 125,000x of intact myelin sheath.

Plate 25. View of spilled cytosol and numerous mitochondria. Ripped up cell membranes are clearly evident.

Plate 28. High power view demonstrating neurofilaments are intact.

General comment. Most damage is associated with lipid bilayers. Double membranes such as nuclear membranes, and complex membranes such as in mitochondria, seem more resistant to damage.

Set 3. Plate 39. Demonstration of some intact membranes from the tissue sample immediately below the outermost tissue sample. This sample is 0.5 cm down from the brain surface. Mitochondria and a portion of a Golgi are illustrated. A portion of intact cell membrane is present. Free ribosomes are seen at the top of the picture and are probably from the adjacent cell.

FINAL COMMENTS. In all samples at all magnifications, simple membranes are the most prone to damage, followed by endoplasmic reticulum, Golgi and mitochondria. Ribosomes and neurofilaments appear resistant. Blood vessels are often intact presumably due to their particular basal lamina.

The outermost sample has the most damage, possibly due to freezing/thawing, followed by the inner-most sample. Intermediate regions had less damage than other areas.

The following photos are plates 28 and 39,which are commented upon in the text above. I don't know how well they will survive the processes of scanning, transmission, and printing.  

Photomicrogram 28-Click for a larger view.

Photomicrogram 39-Click for a larger view.