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<div class="moz-cite-prefix">Hi Christopher,<br>
<br>
this is a rather theoretical reply after looking into available
literature which you surely also consulted (e.g. Butters et al
2014 & Balint et al 2016 on Plos One).<br>
<br>
Both papers basically say that the staining results are hard to
predict and largely dependent on perfusion rates and tissue
thickness (which of course is no surprise but pure chemistry and
physics). Obvious issues effecting the scan results are
over-staining or under-staining, and the optimal incubation time
is hard to predict and depends on specimens size, tissue
composition and - surely too - deposits in and on the specimens
(the cholesterol you already noticed surely is one of the most
obvious ones).<br>
<br>
Other influencing factor are is the x-ray source and data
recovery.<br>
<br>
Personally (gut feeling rather then proofed experience), exposure
to water based solutions for weeks or even months surely damages
the specimens and weakens the tissues, and my next question would
be what happens will all those (dissolved) fats in the specimens
if they are immersed in water for such a long time. Also, Balint
et al (2016) report that over-staining can lead to deformation and
tissue shrinkage .<br>
<br>
The other method you mention, exposure to nearly 100% EtOH has
just the effects that you describe, and the question for me is
what would happen especially to those tissues near the body
surface (e.g. in fish increasing the EtOH strength may cause cell
rupture in the scale pockets and consequently may loosen the fish
scales).<br>
<br>
So it might be worth to explore two alternatives:<br>
<br>
1. It might be worth to compare the imaging capacities of
different ct-scanners; colleagues at the MNHN Paris for example
used the synchroton in Geneva for imaging of or Lungfish larvae
with amazing results.<br>
<br>
2. You mentioned that targeted specimens in your collection were
preserved +100 yrs, some probably used pre-formalin fixatives. If
this is the case, it might be worth to test if any mercury salts
have been added. Balint et al (2016) mention, that "Mercury (II)
chloride provide the best contrast between different soft tissue
types".<br>
<br>
Around 1900, Mercury salts were not rarely added to enhance the
"fixing" properties of the ethanol. Perhaps some of your specimens
have already been treated with a contrast enhancing agent?<br>
<br>
Hope this helps, even though it is not a direct answer to your
question.<br>
All the best<br>
Dirk <br>
<br>
<br>
<br>
Am 30.06.2017 um 19:59 schrieb Milensky, Christopher:<br>
</div>
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<p class="MsoNormal">Hello,<o:p></o:p></p>
<p class="MsoNormal">I am hoping that some folks in the NHCOLL
community have experience with iodine staining and would be
willing to share their knowledge. We are receiving more and
more requests to use our fluid specimens (birds) for CT
scanning projects, some of which require staining. In the
recent past, we have been asked to approve two different
staining techniques. One is a ‘water’ based staining method<span
style="color:#1F497D">
</span>and the other an ‘ethanol’ based method. We would like
to hear opinions about which method is best for the short and
long term care of the specimens. As with any old collection,
the exact methods of fixation used over the last 100+ years
have been variable, but typically our birds were fixed in
formaldehyde and then moved to storage in 70% ETOH. I’m sure
some of our oldest specimens were never properly ‘fixed’ since
formaldehyde did not come into use until the early 1900’s.<o:p></o:p></p>
<p class="MsoNormal"><o:p> </o:p></p>
<p class="MsoNormal">The two methods:<o:p></o:p></p>
<p class="MsoNormal"><o:p> </o:p></p>
<p><span
style="font-family:"Calibri",sans-serif;color:black">I2KI
(potassium iodine and iodide) is dissolved in water and
makes a naturally antimicrobial solution in which the
specimen is removed from ETOH storage and soaked for weeks
to months, depending on the size of the animal, in this
water solution. Pickled tissues (fixed in formalin and
preserved in 70% ethanol) should remain pickled even after
being</span><span
style="font-family:"Calibri",sans-serif;color:#1F497D">
</span><span
style="font-family:"Calibri",sans-serif;color:black">removed
from preservative due to the chemical changes that occur
during fixation, and iodine prevents bacterial growth, but
it still involves putting a pickled specimen in water for an
extended period. <o:p></o:p></span></p>
<p><span
style="font-family:"Calibri",sans-serif;color:black"><o:p> </o:p></span></p>
<p><span
style="font-family:"Calibri",sans-serif;color:black">I2E
is elemental iodide dissolved in 100% ethanol (200 proof),
which raises another set of issues for specimens stored in
lower concentrations of ethanol. Increasing the
concentration of ethanol further dehydrates and shrinks the
specimen, but it has been found to be the most effective
concentration of ethanol for staining soft tissues.<o:p></o:p></span></p>
<p><span
style="font-family:"Calibri",sans-serif;color:black"><o:p> </o:p></span></p>
<p><span
style="font-family:"Calibri",sans-serif;color:black">So,
the question is: Would you rather have a specimen removed
from 70% and stored in a water/iodine solution for
weeks/months or stored in 100% ETOH for a similar period or
neither? What are the pro/cons of each? Would you allow a
specimen collected pre-1910, and presumably not fixed, go
into water/iodine for a month or more? Are you aware of
other methods?<o:p></o:p></span></p>
<p><span
style="font-family:"Calibri",sans-serif;color:black"><o:p> </o:p></span></p>
<p><span
style="font-family:"Calibri",sans-serif;color:black">I
look forward to hearing your thoughts!<o:p></o:p></span></p>
<p><span
style="font-family:"Calibri",sans-serif;color:black"><o:p> </o:p></span></p>
<p><span
style="font-family:"Calibri",sans-serif;color:black">Chris<o:p></o:p></span></p>
<p class="MsoNormal"><o:p> </o:p></p>
<p class="MsoNormal"><o:p> </o:p></p>
<p class="MsoNormal"><b><span
style="font-size:9.0pt;font-family:"Arial",sans-serif;color:#1F497D">Christopher
Milensky</span></b><span
style="font-size:9.0pt;font-family:"Arial",sans-serif;color:#1F497D"><o:p></o:p></span></p>
<p class="MsoNormal"><span
style="font-size:9.0pt;font-family:"Arial",sans-serif;color:#1F497D">Collections
Manager<o:p></o:p></span></p>
<p class="MsoNormal"><span
style="font-size:9.0pt;font-family:"Arial",sans-serif;color:#1F497D"><a href="https://urldefense.proofpoint.com/v2/url?u=http-3A__vertebrates.si.edu_birds_&d=DwMFAg&c=cjytLXgP8ixuoHflwc-poQ&r=LpYc_Z_iN1KRw0hheb3x6-8MJUMu482qfHowpGYJqwc&m=_ng57dMSlpRtwx104LoLynttAJTY-hHusbDzZvPHtcM&s=GUIwF2jcK-a5sAF-jpiFUFCe2_UmI54Q-akmUeSa6xE&e=" moz-do-not-send="true"><span style="color:blue">Division
of Birds</span></a><o:p></o:p></span></p>
<p class="MsoNormal"><b><span
style="font-size:9.0pt;font-family:"Arial",sans-serif;color:#1F497D">w</span></b><span
style="font-size:9.0pt;font-family:"Arial",sans-serif;color:#1F497D">
202.633.0794
<a href="mailto:milenskyc@si.edu" moz-do-not-send="true">milenskyc@si.edu</a><o:p></o:p></span></p>
<p class="MsoNormal"><span
style="font-size:9.0pt;font-family:"Arial",sans-serif;color:#1F497D"><o:p> </o:p></span></p>
<p class="MsoNormal"><span
style="font-size:9.0pt;font-family:"Arial",sans-serif;color:#00C1D5">SMITHSONIAN
INSTITUTION<o:p></o:p></span></p>
<p class="MsoNormal"><span
style="font-size:9.0pt;font-family:"Arial",sans-serif;color:#00778B">NATIONAL
MUSEUM OF NATURAL HISTORY<o:p></o:p></span></p>
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