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Hi Qiu-Xing,
<div>I agree totally with Paul. HPF is the appropriate Methode to use. Beside I embedded long time ago several pancreas samples using 2.5% glutharaldehyde in 0.1M caccodylic buffer pH 7.1. Fixation was performed at +4*C over night. This worked quite well for
pancreas as well as for langerhans islands.</div>
<div>Regards</div>
<div>Dietmar</div>
<div>———————————/——</div>
<div>Riedel Dietmar </div>
<div>MPI for biophys. Chemistry</div>
<div>Göttingen</div>
<div><br>
<div id="AppleMailSignature">Mobil gesendet</div>
<div><br>
Am 17.10.2017 um 15:36 schrieb Paul Verkade <<a href="mailto:P.Verkade@bristol.ac.uk">P.Verkade@bristol.ac.uk</a>>:<br>
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<div class="" style="word-wrap:break-word">Hi Qiu-Xing,
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<div class="">That is indeed a known problem. Have you tried High Pressure Freezing followed by freeze substitution? This worked very well for us, see attached paper:</div>
<div class="">Do let me know if you have any questions about the procedure.</div>
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<div class="">Best regards,</div>
<div class="">Paul<br class="">
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<div class="">__________________________________<br class="">
Paul Verkade PhD FRMS<br class="">
Professor of Bioimaging<br class="">
Wolfson Bioimaging Facility<br class="">
School of Biochemistry<br class="">
Biomedical Sciences Building<br class="">
University of Bristol<br class="">
University Walk<br class="">
BS8 1TD Bristol<br class="">
United Kingdom<br class="">
<br class="">
tel.+44-117-3312179<br class="">
Fax. +44-117-3312168<br class="">
mail: <a href="mailto:p.verkade@bristol.ac.uk" class="">p.verkade@bristol.ac.uk</a><br class="">
websites:<br class="">
<a href="http://www.bristol.ac.uk/biochemistry/research/pv.html" class="">http://www.bristol.ac.uk/biochemistry/research/pv.html</a><br class="">
<a href="http://www.bristol.ac.uk/biochemistry/wbif" class="">http://www.bristol.ac.uk/biochemistry/wbif</a><br class="">
<br class="">
A top 5 UK university with leading employers (2015)<br class="">
A top 5 UK university for research (2014 REF)<br class="">
A world top 40 university (QS Ranking 2015)<br class="">
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<div class="">On 17 Oct 2017, at 13:19, Jiang,Qiu-Xing <<a href="mailto:qxjiang@ufl.edu" class="">qxjiang@ufl.edu</a>> wrote:</div>
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<div class="">Dear all, </div>
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<div class="">Recently scientists at our facility used a standard fixation/embedding protocol to process insulin-secreting cells in order to preserve the secretory granules (SGs) inside them. Even though most intracellular organelles and structures are pretty
well preserved, the preservation of the SG's was very poor and almost all granules were lost, leaving empty space filled with embedding resin. The SG's are known to have very tight membranes, which likely caused the problem. While different protocols are being
tested, I am writing to seek some advice from those who might have encountered similar technical issues, and are willing to share their solutions. If so, please email me at<span class="x_Apple-converted-space"> </span><a href="mailto:qxjiang@ufl.edu" class="" style="color:purple; text-decoration:underline">qxjiang@ufl.edu</a>.
Thanks in advance. </div>
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Qiu-Xing</div>
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<div class="" style="margin:0in 0in 0.0001pt; font-size:11pt; font-family:'Times New Roman',serif">
<b class=""><span class="" style="font-size:16pt; color:rgb(47,84,150)">Qiu-Xing Jiang, Ph.D.</span></b></div>
<div class="" style="margin:0in 0in 0.0001pt; font-size:11pt; font-family:'Times New Roman',serif">
<i class=""><span class="" style="font-size:12pt">Department Microbiology & Cell Science at UF IFAS </span></i><i class=""><span class="" style="font-size:12pt"></span></i></div>
<div class="" style="margin:0in 0in 0.0001pt; font-size:11pt; font-family:'Times New Roman',serif">
<i class=""><span class="" style="font-size:12pt">Electron Microscopy Faculty Director at UF ICBR</span></i></div>
<div class="" style="margin:0in 0in 0.0001pt; font-size:11pt; font-family:'Times New Roman',serif">
PO Box 110700 | 1355 Museum Drive, Rms 1298 or 1003, Gainesville, FL 32611-0700 USA</div>
<div class="" style="margin:0in 0in 0.0001pt; font-size:11pt; font-family:'Times New Roman',serif">
<a href="https://www.biotech.ufl.edu/" class="" style="color:rgb(149,79,114); text-decoration:underline">Biotech.ufl.edu</a> | Tel: 352.846.0953| Fax: 352.392.5933 |E-mail: <a href="mailto:qxjiang@ufl.edu" class="" style="color:rgb(149,79,114); text-decoration:underline">qxjiang@ufl.edu</a></div>
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<b class="" style="font-size:11pt"><span class="" style="font-size:16pt; color:rgb(31,78,121)"><span id="x_cid:177CEC07-6758-4C0D-9153-30A2D3EA1141"><0D571167-8F2C-44BC-A5F0-D789C9297B6C[64].png></span></span></b></div>
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<div><2012-Fava-Diabetologia.pdf></div>
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