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<div class="moz-cite-prefix">.. and often herpetology specimens are only injected, but not tossed into fixatives or (pseudo)fixatives; might be another key factor to consider in this specific case ....
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<div class="moz-cite-prefix">Am 07.11.2023 um 12:28 schrieb <a class="moz-txt-link-abbreviated" href="mailto:a.j.van_dam@lumc.nl">
a.j.van_dam@lumc.nl</a>:<br>
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<p class="MsoNormal"><span style="mso-fareast-language:EN-US" lang="EN-GB">Dear Kelly,
<o:p></o:p></span></p>
<p class="MsoNormal"><span style="mso-fareast-language:EN-US" lang="EN-GB"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="mso-fareast-language:EN-US" lang="EN-GB">The most stable organic pigments (melanins) are found in keratin.
</span><span lang="EN-US">This is well demonstrated by the direct detection of melanin in dinosaur fossils.
</span><span style="mso-fareast-language:EN-US" lang="EN-GB">These pigments in keratin-rich tissue like (snake) skin and hairs can sustain quite a long time when preserved in ethanol. However, the other tissues of the specimen will lose their colour much faster
and become pale/whitish in appearance.<o:p></o:p></span></p>
<p class="MsoNormal"><span style="mso-fareast-language:EN-US" lang="EN-GB"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="mso-fareast-language:EN-US" lang="EN-GB">Besides its flammability and high vapor pressure, another disadvantage of ethanol is that it dissolves lipids and extracts fat soluble organic piments (like carotenoids), leading to
loss of tissue colour and yellowing of the fluid. Using glycerol 65% as an alternative solves these two problems. Glycerol has extreme low vapor pressure and does not dissolve lipids due to its high polarity.
<o:p></o:p></span></p>
<p class="MsoNormal"><span style="mso-fareast-language:EN-US" lang="EN-GB"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="mso-fareast-language:EN-US" lang="EN-GB">With regard to the observed flaccidness, it might be possible that in this case fixation was only performed with ethanol (pseudo) fixation.<o:p></o:p></span></p>
<p class="MsoNormal"><span style="mso-fareast-language:EN-US" lang="EN-GB"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="mso-fareast-language:EN-US" lang="EN-GB">Kind regards,<o:p></o:p></span></p>
<p class="MsoNormal"><span style="mso-fareast-language:EN-US" lang="EN-GB"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="mso-fareast-language:EN-US" lang="EN-GB">Dries<o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-US"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="font-size:8.0pt;font-family:"Verdana",sans-serif;color:#0070C0"><a href="https://www.linkedin.com/profile/preview?vpa=pub&locale=en_US" moz-do-not-send="true"><span style="color:#0070C0;text-decoration:none">Andries J. van
Dam</span></a></span><span style="font-size:8.0pt;font-family:"Verdana",sans-serif;color:#7F7F7F"> | curator-conservator</span><span style="color:#7F7F7F"><o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:8.0pt;font-family:"Verdana",sans-serif;color:#7F7F7F"> </span><span style="color:#7F7F7F"><o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:8.0pt;font-family:"Verdana",sans-serif;color:#0070C0"><a href="https://www.lumc.nl/onderwijs/over-ons/anatomisch-museum/?setlanguage=English&setcountry=en" moz-do-not-send="true"><span style="color:#0070C0;text-decoration:none" lang="EN-US">Anatomical
Museum</span></a></span><span style="font-size:8.0pt;font-family:"Verdana",sans-serif;color:#7F7F7F" lang="EN-US"> | Directorate of education
</span><span style="font-size:8.0pt;font-family:"Verdana",sans-serif;color:#7F7F7F" lang="EN-US">|</span><span style="font-size:8.0pt;font-family:"Verdana",sans-serif;color:#7F7F7F" lang="EN-US"> Leiden University Medical Center | Building 3 (V3-32)</span><span style="color:#7F7F7F" lang="EN-US"><o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:8.0pt;font-family:"Verdana",sans-serif;color:#7F7F7F" lang="EN-US">P.O.Box 9600 | 2300 RC Leiden | Netherlands</span><span style="color:#7F7F7F" lang="EN-US"><o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:8.0pt;font-family:"Verdana",sans-serif;color:#7F7F7F" lang="EN-US">Visiting address: Hippocratespad 21 | Tel: +31 (0)71 52 68356 | E-mail:
</span><span style="color:#0070C0"><a href="mailto:A.J.van_Dam@lumc.nl" target="_blank" moz-do-not-send="true"><span style="font-size:8.0pt;font-family:"Verdana",sans-serif;color:#0070C0;text-decoration:none" lang="EN-US">A.J.van_Dam@lumc.nl</span></a></span><span style="color:#7F7F7F" lang="EN-US"><o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size:8.0pt;font-family:"Verdana",sans-serif;color:#7F7F7F" lang="EN-US"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="font-size:8.0pt;font-family:"Verdana",sans-serif;color:#7F7F7F" lang="EN-US">Scientific associate | Natural History Museum London</span><span style="color:#0070C0" lang="EN-US"><o:p></o:p></span></p>
<p class="MsoNormal"><span lang="EN-US"><o:p> </o:p></span></p>
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<p class="MsoNormal"><span style="mso-fareast-language:EN-US" lang="EN-GB"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="mso-fareast-language:EN-US" lang="EN-GB"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="mso-fareast-language:EN-US" lang="EN-GB"><o:p> </o:p></span></p>
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<p class="MsoNormal"><b><span lang="EN-US">From:</span></b><span lang="EN-US"> Nhcoll-l
<a class="moz-txt-link-rfc2396E" href="mailto:nhcoll-l-bounces@mailman.yale.edu">
<nhcoll-l-bounces@mailman.yale.edu></a> <b>On Behalf Of </b>John E Simmons<br>
<b>Sent:</b> maandag 6 november 2023 22:41<br>
<b>To:</b> Cassidy, Kelly Michela <a class="moz-txt-link-rfc2396E" href="mailto:cassidyk@wsu.edu">
<cassidyk@wsu.edu></a><br>
<b>Cc:</b> <a class="moz-txt-link-abbreviated" href="mailto:nhcoll-l@mailman.yale.edu">
nhcoll-l@mailman.yale.edu</a><br>
<b>Subject:</b> Re: [Nhcoll-l] Speculation on color retention and color loss in preserved herps?<o:p></o:p></span></p>
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<p class="MsoNormal"><o:p> </o:p></p>
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<p class="MsoNormal" style="line-height:106%"><span style="font-size:10.0pt;line-height:106%;font-family:"Arial",sans-serif;color:black">Kelly,</span><span style="font-size:12.0pt;line-height:106%;color:black"><o:p></o:p></span></p>
<p class="MsoNormal" style="line-height:106%"><span style="font-size:10.0pt;line-height:106%;font-family:"Arial",sans-serif;color:black">Color change in preserved specimens is a complex problem, as I discussed at some length in
<i>Fluid Preservation: A Comprehensive Reference</i> (2014) along with a review the available literature on the subject. In short, changes of color may result from chemical or physical alterations of the tissues (usually both) and may involve color loss, acquisition,
or alteration. Of the factors that cause color change, the most significant are light UV exposure, shrinkage, and swelling. For example, in reptiles and amphibians, yellows and greens tend to fade quickly, but blacks and browns are more stable. The common
green-to-blue change in herps is largely (but not exclusively) due to the leaching of xanthophories by the preservative and the alteration of iridophores by dehydration.</span><span style="font-size:12.0pt;line-height:106%;font-family:"Tahoma",sans-serif;color:black"><o:p></o:p></span></p>
<p class="MsoNormal" style="line-height:106%"><span style="font-size:10.0pt;line-height:106%;font-family:"Arial",sans-serif;color:black"> </span><span style="font-size:12.0pt;line-height:106%;font-family:"Tahoma",sans-serif;color:black"><o:p></o:p></span></p>
<p class="MsoNormal" style="line-height:106%"><span style="font-size:10.0pt;line-height:106%;font-family:"Arial",sans-serif;color:black">Color in amphibians and reptiles involves pigments (some of which are soluble in preservatives) and light reflection and
refraction, which is altered by the shrinkage and swelling of tissues. Color change may begin with post-mortem specimens handling (e.g., the time interval between death and fixation or preservation, freezing the specimen, exposure to sunlight). Once the specimen
is preserved, both visible light and ultraviolet radiation will alter colors, as will heat. The choice of preservatives is also a factor—colors will change in different ways if the specimen is in formaldehyde, ethanol, or isopropanol. The colors of the specimen
may be affected by dyes from the string, tags, labels, or container, and exposure to oxidation processes, and exposure to metals (particularly copper).</span><span style="font-size:12.0pt;line-height:106%;font-family:"Tahoma",sans-serif;color:black"><o:p></o:p></span></p>
<p class="MsoNormal" style="line-height:106%"><span style="font-size:10.0pt;line-height:106%;font-family:"Arial",sans-serif;color:black"> </span><span style="font-size:12.0pt;line-height:106%;font-family:"Tahoma",sans-serif;color:black"><o:p></o:p></span></p>
<p class="MsoNormal" style="line-height:106%"><span style="font-size:10.0pt;line-height:106%;font-family:"Arial",sans-serif;color:black">The preservation literature is full of magic recipes to preserve color, none of which work. The only universal among the
recipes is that I have never seen an article in which the colors of the live specimen and the post-preservation specimen were compared to the same color standard, which means that all estimations of how well the colors were preserved were biased guesses.</span><span style="font-size:12.0pt;line-height:106%;font-family:"Tahoma",sans-serif;color:black"><o:p></o:p></span></p>
<p class="MsoNormal" style="line-height:106%"><span style="font-size:10.0pt;line-height:106%;font-family:"Arial",sans-serif;color:black"> </span><span style="font-size:12.0pt;line-height:106%;font-family:"Tahoma",sans-serif;color:black"><o:p></o:p></span></p>
<p class="MsoNormal" style="line-height:106%"><span style="font-size:10.0pt;line-height:106%;font-family:"Arial",sans-serif;color:black">The photo that you included is interesting—that is, indeed, a spectacular retention of colors by a snake that has been in
preservative (and probably exposed to light on view?) since 1938. Typically, that level of pattern retention is only visible on specimens kept largely in the dark (thus protected from visible light and UV). The snake has lost its yellow-green-red colors (most
of which are alcohol soluble pigments), but otherwise looks good. Are there any notes in your institutional archives or collectors field books that might tell you what technique was used to preserve the snake? There were a lot of different chemicals in use
for preservation around that time.</span><span style="font-size:12.0pt;line-height:106%;font-family:"Tahoma",sans-serif;color:black"><o:p></o:p></span></p>
<p class="MsoNormal" style="line-height:106%"><span style="font-size:10.0pt;line-height:106%;font-family:"Arial",sans-serif;color:black"> </span><span style="font-size:12.0pt;line-height:106%;font-family:"Tahoma",sans-serif;color:black"><o:p></o:p></span></p>
<p class="MsoNormal" style="line-height:106%"><span style="font-size:10.0pt;line-height:106%;font-family:"Arial",sans-serif;color:black">Keeping the true (live) color of specimens preserved in fluid is the Holy Grail of fluid preservation, as can be seen from
the numerous publications on the subject suggesting ways to do it. Unfortunately, all of the published magic recipes fail the color standard test. If colors of the specimen are important, it should be photographed before it is preserved with a color standard
reference in the photo so the colors in the image can be corrected later.</span><span style="font-size:12.0pt;line-height:106%;font-family:"Tahoma",sans-serif;color:black"><o:p></o:p></span></p>
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<p class="MsoNormal" style="line-height:106%"><span style="font-size:10.0pt;line-height:106%;font-family:"Arial",sans-serif;color:black">--John</span><span style="font-size:12.0pt;line-height:106%;font-family:"Tahoma",sans-serif;color:black"><o:p></o:p></span></p>
<p class="MsoNormal" style="line-height:106%"><span style="font-size:12.0pt;line-height:106%;font-family:"Tahoma",sans-serif;color:black"><o:p> </o:p></span></p>
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<p class="MsoNormal"><span style="font-size:10.0pt;font-family:"Arial",sans-serif">John E. Simmons</span><span style="font-size:10.0pt;font-family:"Tahoma",sans-serif"><br>
Writer and Museum Consultant</span><o:p></o:p></p>
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<p class="MsoNormal"><span style="font-size:10.0pt;font-family:"Tahoma",sans-serif">Museologica<br>
<i>and</i><br>
Investigador Asociado, Departamento de Ornitologia<br>
Museo de Historia Natural, Universidad Nacional Mayor de San Marcos, Lima</span><o:p></o:p></p>
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<p class="MsoNormal">On Mon, Nov 6, 2023 at 3:04 PM Cassidy, Kelly Michela <<a href="mailto:cassidyk@wsu.edu" moz-do-not-send="true" class="moz-txt-link-freetext">cassidyk@wsu.edu</a>> wrote:<o:p></o:p></p>
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<p class="MsoNormal" style="mso-margin-top-alt:auto;mso-margin-bottom-alt:auto"><span lang="EN-US">Curious about herp preservation techniques that affect color retention and loss.
<o:p></o:p></span></p>
<p class="MsoNormal" style="mso-margin-top-alt:auto;mso-margin-bottom-alt:auto"><span lang="EN-US"> <o:p></o:p></span></p>
<p class="MsoNormal" style="mso-margin-top-alt:auto;mso-margin-bottom-alt:auto"><span lang="EN-US">We have a garter snake that was once used as a display and/or teaching specimens. It was in a straight glass tube, sealed at the ends with a black tarry substance.
About 10 or 15 years ago, I removed the snake from its glass tube because the fluid and glass had become too discolored for it to be a suitable display specimen. I rinsed it and transferred it to a jar in 70% ethanol, but I am not sure what the fluid in the
original tube was. The snake was collected in 1938. For a specimen approaching 90 years old, its color pattern was unusually sharp, but it is also much more flaccid than a typical snake fixed in formalin and preserved in ethanol. (Picture attached.) Any idea
what fixative or storage chemicals might have caused better color retention but might have been less good at preservation, leading to more flaccidness?
<o:p></o:p></span></p>
<p class="MsoNormal" style="mso-margin-top-alt:auto;mso-margin-bottom-alt:auto"><span lang="EN-US"> <o:p></o:p></span></p>
<p class="MsoNormal" style="mso-margin-top-alt:auto;mso-margin-bottom-alt:auto"><span lang="EN-US"> <o:p></o:p></span></p>
<p class="MsoNormal" style="mso-margin-top-alt:auto;mso-margin-bottom-alt:auto"><span lang="EN-US">On the other end of spectrum, we have a number of specimens, most from the mid-20<sup>th</sup> century (1950s to 1970s) that are now almost entirely bleached
of color. These nearly white specimens came to Conner Museum as part of an “orphaned” collection (from Walla Walla College). Not all of the specimens from Walla Walla are bleached out. I am guessing there was a period when their fixative or storage solution
contained or lacked something that caused unusual bleaching. We have no records from the collection about fixative or storage methods. What is the most likely cause of such extreme bleaching? Lack of buffering or chemicals used for buffering? Storage in denatured
ethanol? <o:p></o:p></span></p>
<p class="MsoNormal" style="mso-margin-top-alt:auto;mso-margin-bottom-alt:auto"><span lang="EN-US"> <o:p></o:p></span></p>
<p class="MsoNormal" style="mso-margin-top-alt:auto;mso-margin-bottom-alt:auto"><span lang="EN-US"> <o:p></o:p></span></p>
<p class="MsoNormal" style="mso-margin-top-alt:auto;mso-margin-bottom-alt:auto"><span lang="EN-US">Dr. Kelly M. Cassidy, Curator, Conner Museum<o:p></o:p></span></p>
<p class="MsoNormal" style="mso-margin-top-alt:auto;mso-margin-bottom-alt:auto"><span lang="EN-US">School of Biological Sciences<o:p></o:p></span></p>
<p class="MsoNormal" style="mso-margin-top-alt:auto;mso-margin-bottom-alt:auto"><span lang="EN-US">Box 644236<o:p></o:p></span></p>
<p class="MsoNormal" style="mso-margin-top-alt:auto;mso-margin-bottom-alt:auto"><span lang="EN-US">Washington State University<o:p></o:p></span></p>
<p class="MsoNormal" style="mso-margin-top-alt:auto;mso-margin-bottom-alt:auto"><span lang="EN-US">Pullman, WA 99164-4236<o:p></o:p></span></p>
<p class="MsoNormal" style="mso-margin-top-alt:auto;mso-margin-bottom-alt:auto"><span lang="EN-US">509-335-3515<o:p></o:p></span></p>
<p class="MsoNormal" style="mso-margin-top-alt:auto;mso-margin-bottom-alt:auto"><span lang="EN-US"> <o:p></o:p></span></p>
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<pre class="moz-quote-pre" wrap="">_______________________________________________
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<b><span style="font-family:"Source Sans Pro",sans-serif" lang="EN-US">Dirk Neumann</span></b></p>
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normal">
<span style="font-family:"Source Sans Pro",sans-serif" lang="EN-US">Collection Manager, Hamburg</span></p>
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normal">
<span style="font-family:"Source Sans Pro",sans-serif" lang="EN-US"> </span></p>
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<span style="font-family:"Source Sans Pro",sans-serif" lang="EN-US">Postal address:</span></p>
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normal">
<b><span style="font-family:"Source Sans Pro",sans-serif" lang="EN-US">Museum of Nature Hamburg</span></b><span style="font-family:"Source Sans Pro",sans-serif" lang="EN-US"><br>
</span><span style="font-family:"Source Sans Pro",sans-serif;
color:#008669" lang="EN-US">Leibniz Institute for the Analysis</span></p>
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normal">
<span style="font-family:"Source Sans Pro",sans-serif;color:#008669">of Biodiversity Change</span></p>
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<span style="font-family:"Source Sans Pro",sans-serif">Dirk Neumann</span></p>
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normal">
<span style="font-family:"Source Sans Pro",sans-serif">Martin-Luther-King-Platz 3</span></p>
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normal">
<span style="font-family:"Source Sans Pro",sans-serif">20146</span><span style="font-family:"Source Sans Pro",sans-serif">
</span><span style="font-family:"Source Sans Pro",sans-serif">Hamburg</span><span style="font-family:"Source Sans Pro",sans-serif"><br>
+49 </span><span style="font-family:"Source Sans Pro",sans-serif">40</span><span style="font-family:"Source Sans Pro",sans-serif">
</span><span style="font-family:"Source Sans Pro",sans-serif">238 3</span><span style="font-family:"Source Sans Pro",sans-serif">17 –
</span><span style="font-family:"Source Sans Pro",sans-serif">628</span></p>
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<u><span style="font-family:"Source Sans Pro",sans-serif;color:blue"><a href="mailto:d.neumann@leibniz-lib.de" class="moz-txt-link-freetext">d.neumann@leibniz-lib.de</a></span></u></p>
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<a href="imap://dneumann@webmail.leibniz-lib.de:993/fetch%3EUID%3E/INBOX/Privat/www.leibniz-lib.de"><span style="font-family:"Source Sans Pro",sans-serif">www.leibniz-lib.de</span></a></p>
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<span style="font-size:10.0pt;font-family:"Source Sans Pro",sans-serif">-- <br>
Stiftung Leibniz-Institut zur Analyse des Biodiversitätswandels<br>
Postanschrift: Adenauerallee 127, 53113 Bonn, Germany<br>
<br>
Stiftung des öffentlichen Rechts;<br>
Generaldirektion: Prof. Dr. Bernhard Misof (Generaldirektor), Adrian Grüter (Kaufm. Geschäftsführer)<br>
Sitz der Stiftung: Adenauerallee 160 in Bonn<br>
Vorsitzender des Stiftungsrates: Dr. Michael Wappelhorst</span></p>
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"Source Sans Pro",sans-serif"> </span></p>
</div>
</div>
-- <br>
Stiftung Leibniz-Institut zur Analyse des Biodiversitätswandels<br>
Postanschrift: Adenauerallee 127, 53113 Bonn, Germany<br>
<br>
Stiftung des öffentlichen Rechts;<br>
Generaldirektion: Prof. Dr. Bernhard Misof (Generaldirektor), Adrian Grüter (Kaufm. Geschäftsführer)<br>
Sitz der Stiftung: Adenauerallee 160 in Bonn<br>
Vorsitzender des Stiftungsrates: Dr. Michael Wappelhorst<br>
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