Name: do-it-yourself device for recovery of cryopreserved samples accidentally dropped into cryogenic storage tanks
Uploaded: 2012-05-11T13:00:00
Description: Watch this Scientific Journal Video about Do-It-Yourself Device for Recovery of Cryopreserved Samples Accidentally Dropped into Cryogenic Storage Tanks at JoVE.com

DIY cryogenic retrieval device had been tested in course of the projects "Modeling bacterial protein prenylation in eukaryotic cells using bioinformatics and molecular approaches" sponsored by Mary Louise Andrews Award for Cancer Research program, Virginia Academy of Science, "Physiological Meaning of RNA Editing in Plasma Dendritic Cells" sponsored by The Thomas F. Jeffress and Kate Miller Jeffress Memorial Trust and State Contract 16.740.11.0364 (Ministry of Science and Education, Russia). We would like to thank Beth Eom for assistance with the RNA integrity experiment.

1. Assembly of Cryotolerant Device for Retrieval of Cryoboxes
<ol>
 <li>Using pliers, straighten out one side of 3 sided strong-tie (Figure 1A) to make an L shaped strong-tie shown in Figure 1B. Note: The dimensions of strong-tie can be selected depending on the diameter of the neck of Dewar.</li>
 <li>Secure two such strong-ties with a strong-tie T strap (Figure 2) using Crown bolt nut, washer and screw to form the Cryoscoop base as shown in F...

<ol>
	<li>Kittel, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Advances in Cryogenic Engineering. , 41 (1996).</li><li>Edeskuty, F. J., Walter, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Stewart Safety in the handling of cryogenic fluids. , (1996).</li><li>Council, N. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Prudent Practices in the Laboratory: Handling and Management of Chemical Hazards. , (2011).</li><li>Muyal, J., Muyal, V., Kaistha, B., Seifart, C., Fehrenbach, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Systematic+comparison+of+RNA+extraction+techniques+from+frozen+and+fresh+lung+tissues:+checkpoint+towards+gene+expression+studies.">Systematic comparison of RNA extraction techniques from frozen and fresh lung tissues: checkpoint towards gene expression studies.</a> Diagnostic Pathology. 4, 9 (2009).</li><li>Sambrook, J., Russell, D. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Molecular cloning: a laboratory manual. , (2001).</li><li>Holland, N. T., Smith, M. T., Eskenazi, B., Bastaki, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Biological+sample+collection+and+processing+for+molecular+epidemiological+studies.">Biological sample collection and processing for molecular epidemiological studies.</a> Mutation Research/Reviews in Mutation Research. 543, 217-234 (2003).</li><li>Lynch, P. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=CELL,+TISSUE+AND+ORGAN+CULTURE+|+Storage+and+Cryopreservation.">CELL, TISSUE AND ORGAN CULTURE | Storage and Cryopreservation.</a> Encyclopedia of Rose Science. , 106-111 (2003).</li><li>Crawford, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Plastics Engineering. , (1998).</li><li>Gorenc, B., Tinyou, R., Syam, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Steel designers' handbook. , (1996).</li><li>Harper, C. A., Petrie, E. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Plastics+materials+and+processes:+a+concise+encyclopedia.">Plastics materials and processes: a concise encyclopedia.</a> Wiley-Interscience. , (2003).</li></ol>

One of the most common methods of retrieving cryoboxes and cryovials from the bottom of the Dewar tank is to decant liquid nitrogen into a spare container and pull out the samples remaining in the flask or floating in the decanted liquid nitrogen. This is, however, an unsafe practice that may cause cryogenic burns or asphyxiation due to prolonged exposure to liquid nitrogen vapors2. Other common way of retrieval of fallen cryovials involves use of cryovial tongs3. However, the limited visibility wit...

<table border="1">
 <tbody>
 <tr>
 <td>Name of the reagent</td>
 <td>Company</td>
 <td>Catalogue number</td>
 <td>Comments </td>
 </tr>
 <tr>
 <td>Hillman Group Plated Slotted Flat 1-3/8 x 4 feet; 14 Gauge</td>
 <td>Local Hardware shop</td>
 <td>N/A</td>
 <td>1 unit required</td>
 </tr>
 <tr>
 <td>Crown Bolt Stainless Steel #6 - 32 x 3/8 in. Oval Head Phillips Drive Machine Screw</td>
 <td>Local Hardware shop</td>
 <td>Model # 42281</td>
 <td>4 units required</td>
 </tr>
 <tr>
 <td>Crown Bolt 3/8 in. x 1-1/2 in. Zinc-Plated Steel Fender Washers</td>
 <td>Local Hardware shop</td>
 <td>Model # 20122</td>
 <td>4 units required</td>
 </tr>
 <tr>
 <td>Crown Bolt Zinc Plated #8-32 Machine Screw Nut</td>
 <td>Local Hardware shop</td>
 <td>Model # 18512</td>
 <td>4 units required</td>
 </tr>
 <tr>
 <td>Simpson Strong-Tie BC40Z 4 In.x4 in. Half Base</td>
 <td>Local Hardware shop</td>
 <td>Model # BC40Z</td>
 <td>2 units required</td>
 </tr>
 <tr>
 <td>Simpson Strong-Tie 6 in. x 6 in. T Strap</td>
 <td>Local Hardware shop</td>
 <td>Model # 66T</td>
 <td>1 unit required</td>
 </tr>
 <tr>
 <td>Oneida 6" Mesh Strainer</td>
 <td>Local Grocery shop</td>
 <td>Model # 7859W</td>
 <td>1 unit required</td>
 </tr>
 <tr>
 <td>The Hillman Group Galvanized Hex Nuts 3/8-16</td>
 <td>Local Hardware shop</td>
 <td>Model #:810509</td>
 <td>6 units required</td>
 </tr>
 <tr>
 <td>The Hillman Group Galvanized Hex Bolts 3/8- 16</td>
 <td>Local Hardware shop</td>
 <td>&nbsp;</td>
 <td>4 units required</td>
 </tr>
 <tr>
 <td>Foam for insulation</td>
 <td>Local Hardware shop</td>
 <td>&nbsp;</td>
 <td>1 unit required</td>
 </tr>
 <tr>
 <td>Screw Driver</td>
 <td>Local Hardware shop</td>
 <td>&nbsp;</td>
 <td>1 unit required</td>
 </tr>
 <tr>
 <td>Pliers</td>
 <td>Local Hardware shop</td>
 <td>&nbsp;</td>
 <td>1 unit required</td>
 </tr>
 </tbody>
</table>

do-it-yourself device for recovery of cryopreserved samples accidentally dropped into cryogenic storage tanks

Liquid nitrogen is colorless, odorless, extremely cold (-196 &deg;C) liquid kept under pressure. It is commonly used as a cryogenic fluid for long term storage of biological materials such as blood, cells and tissues 1,2. The cryogenic nature of liquid nitrogen, while ideal for sample preservation, can cause rapid freezing of live tissues on contact - known as 'cryogenic burn'2, which may lead to severe frostbite in persons closely involved in storage and retrieval of samples from Dewars. Additionally, as liquid nitrogen evaporates it reduces the oxygen concentration in the air and might cause asphyxia, especially in confined spaces2.
In laboratories, biological samples are often stored in cryovials or cryoboxes stacked in stainless steel racks within the Dewar tanks1. These storage racks are provided with a long shaft to prevent boxes from slipping out from the racks and into the bottom of Dewars during routine handling. All too often, however, boxes or vials with precious samples slip out and sink to the bottom of liquid nitrogen filled tank. In such cases, samples could be tediously retrieved after transferring the liquid nitrogen into a spare container or discarding it. The boxes and vials can then be relatively safely recovered from emptied Dewar. However, the cryogenic nature of liquid nitrogen and its expansion rate makes sunken sample retrieval hazardous. It is commonly recommended by Safety Offices that sample retrieval be never carried out by a single person. Another alternative is to use commercially available cool grabbers or tongs to pull out the vials3. However, limited visibility within the dark liquid filled Dewars poses a major limitation in their use.
In this article, we describe the construction of a Cryotolerant DIY retrieval device, which makes sample retrieval from Dewar containing cryogenic fluids both safe and easy.

Watch this Scientific Journal Video about Do-It-Yourself Device for Recovery of Cryopreserved Samples Accidentally Dropped into Cryogenic Storage Tanks at JoVE.com

Do-It-Yourself Device for Recovery of Cryopreserved Samples Accidentally Dropped into Cryogenic Storage Tanks

Here we present a low cost, durable cryotolerant device for sample retrieval from Dewar tanks filled with liquid nitrogen. The ease of construction and modular design of the device makes the process of sample retrieval from cryogenic tanks safe and easy.

Liquid nitrogen is colorless, odorless, extremely cold (-196 &deg;C) liquid kept under pressure. It is commonly used as a cryogenic fluid for long term ...

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