X-Message-Number: 17742
From: 
Date: Fri, 12 Oct 2001 04:53:21 EDT
Subject: LN2ed Lichens Experiment Results

Cryonet:

The lichens I collected and transplanted earlier this summer appeared to be 
doing well at the time of my experiment.  I monitored them for several weeks 
before starting.  However, the problem I briefly mentioned in a previous post 
is persistent--determining whether or not they are living/thriving.  They 
grow slow as oak trees.  What I did was submerge a small "collection" in 
liquid nitrogen to see how they would fair on thawing.  The question is to 
determine whether they will reanimate.

There were three groups in all (all of the same species).  Aside from the LN2 
group, I set aside two equal amounts of the lichens as controls right from 
the start.  One group was left as it was (in the soil surrounding the trunk 
of a large potted plant), the other group was placed (with their attached 
dirt/soil base intact) in a Tupperware-like container and covered with its 
lid (these containers are microwaveable and very light weight/fragile--some 
kind of new polymer and available in grocery stores now and made/marketed by 
"Glad"). Prior to inserting that group in the container, I placed several 
dollars worth of pennies in the bottom to form a "pedestal" but also to hold 
a small volume of rubbing alcohol which would not then be able to reach/soak 
the lichens and attached soil.  I boiled the alcohol first and poured it hot 
such that its gases would force out all air/oxygen/CO2 before placing the 
specimens in and putting on the airtight lid.  This effort was to perhaps 
kill and perhaps preserve that control group.

Once that group was prepared, the other LN2 group was submerged in LN2.  
After a week, I retrieved all of them and placed them back in their original 
spots with the potted plant (where one control group always remained).  They 
have been back for a couple of weeks now.  Over the coming weeks, I will 
attempt to determine how they did.  They all looked fine (while slightly 
different in color) on retrieval.  And they look fine now.  It is possible 
that the LN2 or the alcohol group are in fact dead, but it may take months to 
make the determination.

A convenient means to guage/determine change (that I should have thought of 
sooner but can do if I repeat the experiment--and in the mean time will study 
the process) would/will be to repeatedly photograph them with my digital 
camera, and then digitally read the intensity and hue changes from the 
computer over time with new photographing.  In the Hewlett Packard software 
(that came with the HP camera) you can position the monitor's cursor/arrow at 
any spot on a photographic image and have the computer give out very precise 
digital/numerical readings.  I would need to plug the camera in to an 
electrical socket rather than use batteries (which vary in current with age) 
and photograph them in complete darkness (letting the flash provide all the 
light) and have the subjects precisely positioned in the same way each time 
to achieve consistency  (should not be a big issue and I have a small tripod 
for the camera to hold it steady, also a delay timer will set off the shutter 
to avoid motion from me pressing the button).  This way I could quantify 
changes in color (i.e., hue and intensity) of the two "treated" groups to 
compared to the totally untreated, true control group (which is assumed to be 
living).  The color changes over time in the treated group would be 
attributable to decay.  Of course, the comparative photographing would likely 
demonstrate growth or perhaps subtle deterioration.  Either or both ways may 
shorten the waiting period which otherwise could be many months.

There are many many different types of lichens around the world which are all 
basically symbiotic-type arrangements of fungus and algae, and they have 
hugely different forms.  To see a close approximation of my species go to: 

http://www.lichen.com/smallphotos/Cdeformis.jpeg

The tall stalk-like structures are about one-half a centimeter in length 
(about a fifth of an inch).  My samples are a slight variation/mutation from 
this species (i.e., Cladonia deformis) in that the bright red (spore 
producing fruiting bodies) are more of a rust color, but are just as 
remarkable--every bit as much of an "unnatural" color combination in nature 
between the stalk and the fruiting body (no doubt evolved to attract 
animals/inspects).

(Really serious Lichens-Freaks can go to http://www.lichen.com/portraits.html 
for hyperlinks to some good shots of your favorites.)

More later on the experiment (several weeks or months maybe).

D.C. Johnson

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