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<p>Dear Claire,</p>
<p><br>
</p>
<p>I've been working a lot with Spalteholz specimens lately. I'm a
bit surprised that it was transferred it to glycerol, since none
of the chemicals (methyl salicylate nor benzyl benzoate) are
really miscible with than medium (glycerol). Bedsides that as in
ethanol, there is no real absolute glycerol solution. It will
always absorb some water. And there is the whole issue. The
Spalteholz chemicals are absolutely water repellent. In the
official methodology you actually go from "absolute" ethanol to
benzene in two different baths, to extract all of the water
content from the specimen. From there you go to the methyl
salicylate. The whole method is based on optical properties of the
medium. So yes, in theory you should be able to make the whole
clearing possible again. This depends on the state of preservation
of the specimen.</p>
<p>I would be careful with the assumption that your deposits are
lipids. It is more likely that through the wrong transfer
something has interfered with the whole mixture between glycerol
and the methyl salicylate. It could have formed some kind of
emulsion. A picture could help to clarify.</p>
<p><br>
</p>
<p>I recommend you to read Spalteholz's publication, if there is
some kind of translation (maybe some translation tool might help).
Its really interesting and explains very clearly how the whole
methodology works. This could answer your second question.</p>
<p><br>
</p>
<p>I hope this help a bit.</p>
<p><br>
</p>
<p>All the best!</p>
<p><br>
</p>
<p><br>
</p>
<p><br>
</p>
<div class="moz-cite-prefix">Am 05.11.25 um 10:18 schrieb Claire
Smith:<br>
</div>
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cite="mid:LO0P265MB2778831AF560A8C472BF7D6EC1C5A@LO0P265MB2778.GBRP265.PROD.OUTLOOK.COM">
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Hi everyone, </div>
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<br>
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With apologies for cross-posting, I'm looking for some advice
about working with a damaged specimen. </div>
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<br>
</div>
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It's a newborn monkey that was originally an alizarin
transparency, prepared by the Spalteholz technique (<a
class="OWAAutoLink moz-txt-link-freetext"
id="OWAf6042ca3-eb70-cd33-aa2e-734706ad1500"
href="https://doi.org/10.1016/s0940-9602(01)80020-0"
moz-do-not-send="true">https://doi.org/10.1016/s0940-9602(01)80020-0</a>),
with our catalogue listing the preservative as "Methyl
salicylate 5: benzyl benzoate 4".</div>
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<br>
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In 2008 it was apparently in poor condition with crystal
deposition, and it was transferred into 100% glycerol. </div>
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<br>
</div>
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Currently, a lipid layer has formed at the top of the fluid,
which is also cloudy - presumably also with suspended lipids. </div>
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The tissue is no longer transparent. </div>
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<br>
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My questions are: </div>
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<br>
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Is it possible to make the tissues transparent again? (If so,
how?!)</div>
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Is it best to keep the specimen in glycerol, but dilute it to
a more appropriate concentration? </div>
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<br>
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<br>
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<p
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<span style="color: rgb(0, 0, 0);">Many thanks,</span><br>
</p>
<p
style="margin: 0cm; font-family: Calibri, sans-serif; font-size: 11pt;"
class="elementToProof">
Claire</p>
</div>
<div class="elementToProof" id="Signature">
<p
style="background-color: white; margin: 0cm; font-family: Calibri, sans-serif; font-size: 11pt;"
class="elementToProof">
<span style="color: rgb(50, 49, 48);"> </span></p>
<p
style="background-color: white; margin: 0cm; font-family: Calibri, sans-serif; font-size: 11pt;"
class="elementToProof">
<span style="color: rgb(50, 49, 48);">*******</span></p>
<p
style="background-color: white; margin: 0cm; font-family: Calibri, sans-serif; font-size: 11pt;"
class="elementToProof">
<span style="color: rgb(50, 49, 48);"><b>Survey: Fluid
Preservation Methods in Biological Collections</b></span></p>
<p
style="background-color: white; margin: 0cm; font-family: Calibri, sans-serif; font-size: 11pt;"
class="elementToProof">
<span style="color: blue;"><u><a
style="color: blue; margin-top: 0px; margin-bottom: 0px;"
href="https://app.onlinesurveys.jisc.ac.uk/s/reading/fluid-preservation-methods-in-biological-collections"
moz-do-not-send="true" class="moz-txt-link-freetext">https://app.onlinesurveys.jisc.ac.uk/s/reading/fluid-preservation-methods-in-biological-collections</a></u></span></p>
<p
style="background-color: white; margin: 0cm; font-family: Calibri, sans-serif; font-size: 11pt;"
class="elementToProof">
<span style="color: rgb(50, 49, 48);"> </span></p>
<p
style="background-color: white; margin: 0cm; font-family: Calibri, sans-serif; font-size: 11pt;"
class="elementToProof">
<span style="color: rgb(50, 49, 48);">*******</span></p>
<p style="background-color: white; margin: 0cm;"
class="elementToProof"><span
style="font-family: Calibri, sans-serif; font-size: 11pt; color: rgb(50, 49, 48);"><b>Claire
Smith
</b>(she/her), AFHEA </span></p>
<p
style="background-color: white; margin: 0cm; font-family: Calibri, sans-serif; font-size: 11pt;"
class="elementToProof">
<span style="color: rgb(50, 49, 48);"><b>Graduate Teaching
Assistant: </b>Tuesdays and Wednesdays</span></p>
<p
style="background-color: white; margin: 0cm; font-family: Calibri, sans-serif; font-size: 11pt;"
class="elementToProof">
<span style="color: rgb(50, 49, 48);"><b>PhD Researcher: </b>Mondays,
Thursdays, and Fridays</span></p>
<p style="background-color: white; margin: 0cm;"
class="elementToProof"><span
style="font-family: Calibri, sans-serif; font-size: 14.6667px; color: rgb(50, 49, 48);"><b><br>
</b></span></p>
<p style="background-color: white; margin: 0cm;"
class="elementToProof"><span
style="font-family: Calibri, sans-serif; font-size: 14.6667px; color: rgb(50, 49, 48);"><b>Cole
Museum of Zoology</b></span></p>
<p
style="background-color: white; margin: 0cm; font-family: Calibri, sans-serif; font-size: 11pt;"
class="elementToProof">
<span style="color: rgb(50, 49, 48);">University of Reading</span></p>
<p
style="background-color: white; margin: 0cm; font-family: Calibri, sans-serif; font-size: 11pt;"
class="elementToProof">
<span style="color: rgb(50, 49, 48);"><br>
</span><span style="color: blue;"><u><a
style="color: blue; margin-top: 0px; margin-bottom: 0px;"
target="_blank" href="mailto:claire.smith@reading.ac.uk"
moz-do-not-send="true" class="moz-txt-link-freetext">claire.smith@reading.ac.uk</a></u></span><span
style="color: rgb(0, 112, 192);">
</span></p>
<p
style="background-color: white; margin: 0cm; font-family: Calibri, sans-serif; font-size: 11pt;"
class="elementToProof">
<span style="color: blue; background-color: white;"><u><a
style="color: blue; margin-top: 0px; margin-bottom: 0px;"
href="http://www.linkedin.com/in/wetconservatrix"
moz-do-not-send="true">www.linkedin.com/in/wetconservatrix</a></u></span><span
style="color: rgb(0, 112, 192);"><br>
</span><span style="color: black;">Social media:
@wetconservatrix</span></p>
<p
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class="elementToProof">
</p>
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