1. Preparation for the necropsy
<ol>
 <li>Important parts of any gross postmortem examination (gross necropsy) are the history of the animal and the description of the findings.</li>
 <li>A veterinary pathologist reading your histopathology slides may not have seen the animal and is relying on you for the background information.</li>
 <li>Describe exactly what you see before you euthanize the animal. For example, "female mouse; one of five in the cage; C57BL/6N; head tilted to the right" or "brown male rat; strain ...

<ol>
	<li>AVMA. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> AVMA Guidelines on Euthanasia. , (2007).</li><li>Feldman, D. B., Seely, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Necropsy Guide: Rodents and the rabbit. , (1988).</li><li>King, J. M., Dodd, D. C., Roth, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> The Necropsy Book. , (2006).</li><li>Kittel, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Revised+guides+for+organ+sampling+and+trimming+in+rats+and+mice--Part+2.+A+joint+publication+of+the+RITA+and+NACAD+groups.">Revised guides for organ sampling and trimming in rats and mice--Part 2. A joint publication of the RITA and NACAD groups.</a> Exp Toxicol Pathol. 55, 413-431 (2004).</li><li>Komarek, V., Fox, J. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chapter+1.">Chapter 1.</a> The Mouse in Biomedical Research. , 1-22 (2007).</li><li>Morawietz, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Revised+guides+for+organ+sampling+and+trimming+in+rats+and+mice--Part+3.+A+joint+publication+of+the+RITA+and+NACAD+groups.">Revised guides for organ sampling and trimming in rats and mice--Part 3. A joint publication of the RITA and NACAD groups.</a> Exp Toxicol Pathol. 55, 433-449 (2004).</li><li>Popesko, P., Raijtov&#225;, V., Hor&#225;k, 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> Colour Atlas of Anatomy of Small Laboratory Animals. 2, 91-106 (2002).</li><li>Ruehl-Fehlert, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Revised+guides+for+organ+sampling+and+trimming+in+rats+and+mice--part+1.">Revised guides for organ sampling and trimming in rats and mice--part 1.</a> Exp Toxicol Pathol. 55, 91-106 (2003).</li></ol>

The authors are all employees of Charles River's laboratory rodent diagnostic lab, where these services are offered commercially.

Data collection at the end of a study may require a postmortem examination of animals. Describe well what is seen and remember to examine all tissues. These procedures are designed primarily to optimize necropsy and sample collection for infectious disease surveillance, but most are easily adapted to focused investigations of suspected disease incidents or outbreaks in which perhaps only a subset of the procedures would be employed. Many of the procedures are also relevant to necropsies more commonly performed at the ...

<table border="1">
		<tbody>
				<tr>
						<th>Reagent name</th>
						<th>Company</th>
						<th>Catalog number</th>
						<th>Comments</th>
				</tr>
				<tr>
						<td>Cutting board</td>
						<td>Thermo Electron Corp	Cat #36114</td>
						<td>Cat #36114</td>
						<td>&nbsp;</td>
				</tr>
				<tr>
						<td>Small scissors</td>
						<td>ROBOZ</td>
						<td>RS-5910, G204</td>
						<td>23mm blades, 3.5" length, straight</td>
				</tr>
				<tr>
						<td>Medium scissors	</td>
						<td>ROBOZ</td>
						<td>RS-6808, G207</td>
						<td>5"</td>
				</tr>
				<tr>
						<td>Large scissors</td>
						<td>ROBOZ</td>
						<td>RS-6826, G65"</td>
						<td>6.25"</td>
				</tr>
				<tr>
						<td>Forceps-Curved</td>
						<td>ROBOZ</td>
						<td>RS-8254 (M1/21004)</td>
						<td>4.5", serrated, slight curve</td>
				</tr>
				<tr>
						<td>Forceps-Microdissecting</td>
						<td>ROBOZ</td>
						<td>RS-5238</td>
						<td>Hudson-(EWALD)</td>
				</tr>
				<tr>
						<td>Forceps-Tissue Forceps</td>
						<td>ROBOZ</td>
						<td>RS--8160</td>
						<td>Rat tooth</td>
				</tr>
				<tr>
						<td>Flame sterilizer</td>
						<td>Oxford Labware Products</td>
						<td>Bacti-Cinerator #5889-001007</td>
						<td>&nbsp;</td>
				</tr>
				<tr>
						<td>Scalpel</td>
						<td>Cancer Diagnostics, Inc.</td>
						<td>Finger Scalpels #60, Cat#FS0060</td>
						<td>&nbsp;</td>
				</tr>
				<tr>
						<td>Pipettes</td>
						<td>VWR</td>
						<td>Pasteur Pipet 5 3/4", Cat#14672-400</td>
						<td>&nbsp;</td>
				</tr>
				<tr>
						<td>Autoclave bags</td>
						<td>Propper Manufacturing Company</td>
						<td>Sterilizer Bag Paper Pouches Cat#021002(3100923)E09110</td>
						<td>For Pasteur pipettes</td>
				</tr>
				<tr>
						<td>Pipette bulb</td>
						<td>VWR</td>
						<td>Cat#56310-240</td>
						<td>&nbsp;</td>
				</tr>
				<tr>
						<td>Eppendorf tubes	</td>
						<td>Sarstedt</td>
						<td>SafeSeal microtube 2mL, Ref#72.695</td>
						<td>&nbsp;</td>
				</tr>
				<tr>
						<td>Eppendorf tubes	</td>
						<td>Argos</td>
						<td>5mL microtube Cat#T20765-C</td>
						<td>&nbsp;</td>
				</tr>
				<tr>
						<td>Syringe </td>
						<td>BD (Beckton Dickenson)</td>
						<td>1ml-309602 3 ml-300910 5ml-309603 10mL Cat#309604 20mL Cat#309661</td>
						<td>&nbsp;</td>
				</tr>
				<tr>
						<td>Needles</td>
						<td>BD</td>
						<td>26G-309625 18G-305195 </td>
						<td>&nbsp;</td>
				</tr>
				<tr>
						<td>Neutral buffered formalin</td>
						<td>VWR</td>
						<td>20L 10% NBF, Cat#16004-128</td>
						<td>&nbsp;</td>
				</tr>
				<tr>
						<td>Saline</td>
						<td>ThermoScientific</td>
						<td>Buffered Blood Bank Saline, Cat#23-309-178</td>
						<td>&nbsp;</td>
				</tr>
				<tr>
						<td>Trypticase soy broth</td>
						<td>Becton Dickenson</td>
						<td>TSB: Dehydrated, Cat#211825</td>
						<td>&nbsp;</td>
				</tr>
				<tr>
						<td>Phosphate buffered saline</td>
						<td>Sigma Aldrich</td>
						<td>HA Buffer, Cat#P3813-10PAK</td>
						<td>&nbsp;</td>
				</tr>
				<tr>
						<td>Formalin cups</td>
						<td>VWR</td>
						<td>4oz Cat#36318-852 8oz Cat#36318-860 16oz Cat#36318-858</td>
						<td>&nbsp;</td>
				</tr>
				<tr>
						<td>Large formalin cups </td>
						<td>OakRidge Products</td>
						<td>32oz container Cat#0432-1100</td>
						<td>&nbsp;</td>
				</tr>
				<tr>
						<td>Extra large formalin cup</td>
						<td>VWR</td>
						<td>HDPE Multipurpose 160oz container Cat#89038-282</td>
						<td>&nbsp;</td>
				</tr>
				<tr>
						<td>Suture material </td>
						<td>Henry Schein</td>
						<td>Braided silk surgical suture, Ref#100-5000, M766750</td>
						<td>For mouse lung inflation</td>
				</tr>
				<tr>
						<td>Twine</td>
						<td>Staples</td>
						<td>Cat#QUA-46173</td>
						<td>For rat lung inflation</td>
				</tr>
		</tbody>
</table>

diagnostic necropsy and selected tissue and sample collection in rats and mice

There are multiple sample types that may be collected from a euthanized animal in order to help diagnose or discover infectious agents in an animal colony. Proper collection of tissues for further histological processing can impact the quality of testing results. This article describes the conduct of a basic gross examination including identification of heart, liver, lungs, kidneys, and spleen, as well as how to collect those organs. Additionally four of the more difficult tissue/sample collection techniques are demonstrated. Lung collection and perfusion can be particularly challenging as the tissue needs to be properly inflated with a fixative in order for inside of the tissue to fix properly and to enable thorough histologic evaluation. This article demonstrates the step by step technique to remove the lung and inflate it with fixative in order to achieve optimal fixation of the tissue within 24 hours. Brain collection can be similarly challenging as the tissue is soft and easily damaged. This article demonstrates the step by step technique to expose and remove the brain from the skull with minimal damage to the tissue. The mesenteric lymph node is a good sample type in which to detect many common infectious agents as enteric viruses persist longer in the lymph node than they are shed in feces. This article demonstrates the step by step procedure for locating and aseptically removing the mesenteric lymph node. Finally, identification of infectious agents of the respiratory tract may be performed by bacterial culture or PCR testing of nasal and/or bronchial fluid aspirates taken at necropsy. This procedure describes obtaining and preparing the respiratory aspirate sample for bacterial culture and PCR testing.

Watch this Scientific Journal Video about Diagnostic Necropsy and Selected Tissue and Sample Collection in Rats and Mice at JoVE.com

Diagnostic Necropsy and Selected Tissue and Sample Collection in Rats and Mice

This article describes the procedures for conducting a basic postmortem examination of a mouse or rat, and the collection of basic organs, as well as more challenging sample types from for histological, microbiological, and PCR evaluation.

There are multiple sample types that may be collected from a euthanized animal in order to help diagnose or discover infectious agents in an animal ...

diagnostic-necropsy-selected-tissue-sample-collection-rats

Research

JoVE Journal

Biology

73.6K vistas.  Charles River. En este artículo se describen los procedimientos para la realización de un examen post-mortem básicos de un ratón o una rata, y el conjunto de órganos de base, así como más tipos de muestras reto de la evaluación histológica, microbiológicos y PCR.