X-Message-Number: 4927
Date: 28 Sep 95 13:51:44 EDT
From: Mike Darwin <>
Subject: CRYONICS CryoNet #4921 - #4925

I read with amusement the various posts on the straight freezing issue and 
have a few observations to add of my own:

1) Steve Bridge's response was by the far the best and probably most 
representative of the situation as it really is.

2) Saul's response was a pragmatic one and, since I have discussed this 
issue with him many times over the years, I know wherehe  is coming from.  
Sometimes economy in a reply is not an advantage: this was, in my opinion, 
the case with Saul's response.  As a general note, I would say that I have 
known Saul since I was13 years old (met him first face-to-face when I was 
14).  Saul is neither vindictive, arrogant or suffering from NIH disease.  
He has his weak spots, but he is generally one of the more open and 
deliberating people I've met.  Still, it was a lousy response :).

3) What I find far more interesting is Joe Strout's post.  Joe is a 
neurobiology student in San Diego (and has been for sometime now) which is 
about a 2 hour drive from us in Rancho Cucamonga.  Joe is no doubt very 
busy and thus has little time.  However, others even busier than Joe who 
are even further away have taken the time to come out and kick the tires 
and, further,  two of our personnel with very demanding full time careers 
come out regularly to participate in our brain resuscitation and 
cryopreservation research: often staying all week-end with little or no 
sleep only to have to drive back to work on Monday morning.  One person 
even flies in from San Francisco.

4) Joe is in an excellent position to find his own answers.  EM work is not 
terribly expensive (cut out the beer and pizza for a couple of months and 
you can run three tissue blocks) and Joe is in a place where transmission 
electron microscopy and freeze fracture are a high art.  I feel quite 
certain that he could arrange to have tissue run there on an ad hoc basis 
at little or no cost.  Straight freezing brains is easy, and rodents are, 
for now anyway, exept from USDA regulation. I note that I can legally feed 
rats and mice to my snakes, and I further note that cervical dislocation is 
an easy, straightforward and accepted way to humanely kill rodents (I don't 
consider it acceptable personally, but that's not the point).

5) When CI or I have straight frozen brains what we get back after thawing 
is hash.  If you try to look at straight frozen brains in the frozen state 
with freeze substitution you can't tell anything because all the tissue is 
compressed into VERY narrow and densely compacted channels.

Since we don't do EMs in-house I have developed a network of outside 
microscopists to do the work.  I try to fly up when the tissue is scanned 
so that I can point out what I want photographed.  In every case with 
straight-frozen tissue, without prompting during or after the scan, the 
microscopist has asked me if they are looking at a brain homogenate.  For 
the uninitated a tissue homogenate is most effeciently prepared by use of a 
Waring Blender. Just for the hell of it I once put a brain in a blender 
(Waring variable speed, household type) and pushed the low, medium and high 
speed buttons, pausing to collect tissue after each round. I could get 
results which matched my straight frozen EMs reasonably well  on the mid 
range setting.

6) Saul's point was, I think, that it easy to come up with new ideas that 
are cheaper, easier on everybody NOW, and make cryo"preservation" more 
affordable.  The problem here is simple: we have a lousy product to begin 
with.  In fact, until recently no exhaustive and systematic effort was ever 
undertaken by any cryonics organization to even SEE the ultrastructural and 
histological effects of the BEST procedures we have, or think we have.  One 
of my first priorites with BPI was to find the answer to that question 
(broadly) so that *I* knew what I was giving my "customers" and so that 
they knew too.

Now, there are many caveats to this work, and the same can be said of 
straight freezing.  For all I know those narrow black (profoundly 
osmicated) little channels between the icebergs are chock full o' memory 
and identity.  But, aside from theoretical considerations, I haven't got a 
clue.  This may change as  new kinds of light microscopy offer the 
possibility of near EM resolution coupled with the use of ablative 
techniques on blocks of straight frozen tissue so that the channel material 
could be examined a VERY thin layer at time by essentially grinding off 
micron or sub-micron thick areas  of the surface and then looking at what 
is exposed.

By contrast, well glycerolized brains, examined while frozen or when 
thawed, show a wealth of structure and injury and provide a guide for 
progress and homing in on better approaches.  A great deal of time and 
money have gone into evaluating and minimizing ischemic injury and, if you 
are straight freezing because of costs, let me tell you a good part of the 
cost is in recovering and cooling your patient promptly.

So what are the take-home messages here:

a) Many approaches may work. However, we are not creatures of infinite 
resources and we must CHOOSE.  After choosing we must keep our minds open, 
but we must also have criteria in place to instruct us as to when we should 
abandon one approach and go with another.  

b) failure to do a) will mean that you spend all your time doing research 
on other peoples' ideas, of which (others' ideas) you will find an endless 
supply.  Doug Skrecky and his like could keep the entire biomedical 
infracstructure of the Western World busy for the next 50 years.  BTW Doug, 
you ought to take your proposal to Castro.  Sucrose futures and prices are 
at very low levels and, without the USSR to buy sugar from Cuba at 
ridiculously inflated prices, the Cubanm economy is in even more of a 
shambles than it was.  If everybody starts getting mummified with table 
sugar, Castro's worries are over :). Check Fidel out for seed money; its no 
loonier than a lot of other things communists in general and Castro in 
particular have forked out for in the past. 

c) If Joe, or Brook, or Doug want to straight-freeze, mummify or permafrost 
bury people that is fine by me.  I have learned to stop worrying about what 
will damage the cryonics community as a whole since, in any event, it is 
impossible to STOP people from doing what they damn well please short of 
shooting them in the head.  NOTE: this does not mean that such people 
should not be advised of the perceived harm they might do to "cryonics as a 
whole" (whatever THAT means!).

d) A corollary of c) above is that you have to "show your work" (results) 
or be willing to suffer the consequences or experience the benifits of 
others doing it for you.  And pointing out that you didn't do it yourself.  
All I can say here is that I know a number of neurobiologists and TEM 
people who are going to suggest that your results might be the same if you 
first put your patients' brains through blenders: hell, if you do that, you 
won't even have to straight-freeze, you can just pour in the 
cryoprotectant.  My God!!!!!! Why didn't I thinkof that earlier!  
Eureka!!!!!! The answer!  Not only cheap *cryoprotected* freezing, but you 
can pour the results into VERY compact and variably configured containers: 
why you you could even use 5 ml Nunc tubes and store VERY economically with 
a narrow neck dewar. Taylor-Wharton has some dewars on the market that will 
hold 60 liters of LN2 or so for a YEAR!   This is it, this is the answer!  
What fools we've all been! Why didn't we see it sooner!


Mike Darwin


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