Name: repeated blood collection from tail vein of non-anesthetized rats with a vacuum blood collection system
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Description: Watch this Scientific Journal Video about Repeated Blood Collection from Tail Vein of Non-Anesthetized Rats with a Vacuum Blood Collection System at JoVE.com

作者没有作出任何致谢。

所有程序均由华中科技大学 (中国武汉) 动物保护委员会的同济医学院理事会批准。原稿根据到达准备 (动物研究: 报告在体内实验) 指南6。
1. 畜牧业
<ol><li>使用大 (SD) 大鼠 (n = 20, 10 男性, 8 周老, 体重261到291克, 平均 = 272.85 ±9.07 克) 从实验动物中心的华中理工大学和维护鼠按照指南的护理和使用美国国立卫生研究院出版的实验动物 (NIH 出版物 no. 85-23, 修订 1996)</li>
	<li>在标准条件下的房子, 免费获得食物和饮用水。将老鼠放在530厘米的2笼子里, 用木制的床上用品。在每个笼子里放两只老鼠。保持所有的笼子开放的房间环境 (无微隔离或通风锁定)</li>
	<li>确保参与实验协议的人员定期处理大鼠。</li>
	<li>保持室温介于21° c 和23° c 之间。在整个研究过程中, 给出一个正常的盐饮食 (0.3% 氯化钠)。</li>
</ol>2. 采血程序
注: 真空采血系统由双尖针 [22 克 (0.7 mm x 25 mm)]、一个塑料支架和一系列带橡胶塞子的真空管 (2 毫升) 组成 (图 1)。
<ol><li>将大鼠置于塑料抑制器中, 用温水 (20-30 ° c) 水冲洗尾部, 去除任何可见的污垢或粪便。当助手把尾巴放在较长的位置时, 用70% 的乙醇擦拭尾巴, 用纱布清洗, 使静脉清晰。</li>
	<li>插入22克蝴蝶针尖到一个侧面尾静脉 (大约5毫米) 在一个位置大约 2-3 cm 从尾巴的尖端在近似地° (图 2) 的角度。通过将蝶针的橡胶端插入真空采血管 (图 3) 将血液收集到真空收集管中。 注: 最大采血量为1.2 毫升7。</li>
	<li>采血后, 取出针头, 用纱布轻轻按压穿刺部位15-30 秒, 停止血流。然后, 把老鼠从塑料抑制器中释放出来, 然后把老鼠放回笼子里。</li>
	<li>使用含有 EDTA 作为抗凝血剂的管子来收集血浆。轻轻倒置管几次, 混合抗凝血, 并将样品放在冰垂直。</li>
	<li>将血液样品收集管离心于冷冻离心机中, 在 2000 x g 为10分钟分离血浆或血清。</li>
	<li>提取血浆/血清, 注意不要扰乱红白细胞层。要收集血清, 不用抗凝血管。这两种样品可以立即使用, 或储存在-80 ° c, 长达一年。 注: 成功的血液收集被定义为每次获得1.2 毫升血液。最大穿刺时间设置为三试验, 如果总血容量低于1.2 毫升后, 第三次穿刺, 它被定义为一个失败的血液收集。取样持续时间从尾部静脉穿刺到采血后的蝶针摘除。</li>
	<li>
在两个星期内收集两次血液7。
	<ol>
<li>为了检验和验证这种方法的可行性, 在执行这项采血协议前, 培训两名医科学生 (初学者) 2 小时。使用二十只大鼠, 从10只大鼠中抽取2名经验丰富的研究员 (实验组) 和2名医科学生 (初学者组) 的10只老鼠。</li>
	</ol>
</li>
</ol>3. 统计分析
<ol><li>将数据表示为平均值±标准偏差, 并与 SPSS 统计17进行分析。α值为0.05 被选择, 因此 p < 0.05 被认为有统计学意义。</li>
</ol>

<ol>
	<li>Brown, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Blood+collection+from+the+tail+of+a+rat.">Blood collection from the tail of a rat.</a> Lab Anim (NY). 35 (8), 24-25 (2006).</li><li>Parasuraman, S., Raveendran, R., Kesavan, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Blood+sample+collection+in+small+laboratory+animals.">Blood sample collection in small laboratory animals.</a> J Pharmacol Pharmacother. 1 (2), 87-93 (2010).</li><li>Staszyk, C., Bohnet, W., Gasse, H., Hackbarth, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Blood+vessels+of+the+rat+tail:+a+histological+re-examination+with+respect+to+blood+vessel+puncture+methods.">Blood vessels of the rat tail: a histological re-examination with respect to blood vessel puncture methods.</a> Lab Anim. 37 (2), 121-125 (2003).</li><li>Lee, G., Goosens, K. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sampling+blood+from+the+lateral+tail+vein+of+the+rat.">Sampling blood from the lateral tail vein of the rat.</a> J. Vis. Exp. (99), (2015).</li><li>. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> WHO Guidelines on Drawing Blood: Best Practices in Phlebotomy WHO Guidelines Approved by the Guidelines Review Committee. , (2010).</li><li>Kilkenny, C., Altman, D. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Improving+bioscience+research+reporting:+ARRIVE-ing+at+a+solution.">Improving bioscience research reporting: ARRIVE-ing at a solution.</a> Lab Anim. 44 (4), 377-378 (2010).</li><li>Diehl, K. H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+good+practice+guide+to+the+administration+of+substances+and+removal+of+blood,+including+routes+and+volumes.">A good practice guide to the administration of substances and removal of blood, including routes and volumes.</a> J Appl Toxicol. 21 (1), 15-23 (2001).</li><li>Eder, J. M., Cutter, G. 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+new+device+for+collecting+cord+blood.">A new device for collecting cord blood.</a> Obstet Gynecol. 86 (5), 850-852 (1995).</li><li>Reynolds, B. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+a+new+device+for+blood+sampling+in+cats+with+a+vacuum+tube+collection+system+-+plasma+biochemistry,+haematology+and+practical+usage+assessment.">Comparison of a new device for blood sampling in cats with a vacuum tube collection system - plasma biochemistry, haematology and practical usage assessment.</a> J Feline Med Surg. 9 (5), 382-386 (2007).</li><li>Wiwanitkit, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+blood+specimens+from+plain+and+gel+vacuum+blood+collection+tubes.">Comparison of blood specimens from plain and gel vacuum blood collection tubes.</a> J Med Assoc Thai. 84 (5), 723-726 (2001).</li><li>Wollowitz, A., Bijur, P. E., Esses, D., John Gallagher, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Use+of+butterfly+needles+to+draw+blood+is+independently+associated+with+marked+reduction+in+hemolysis+compared+to+intravenous+catheter.">Use of butterfly needles to draw blood is independently associated with marked reduction in hemolysis compared to intravenous catheter.</a> Acad Emerg Med. 20 (11), 1151-1155 (2013).</li></ol>

作者没有透露

本报告介绍了一种易于学习的大鼠血液收集方法。我们的研究结果表明, 这种方法是可行的重复收集大容量的大鼠血。此外, 这种方法可以很容易地掌握与短期学习曲线。这种方法可以在非麻醉大鼠中进行, 只对动物造成最小的压力。
血液可以从不同地点的老鼠身上收集。尾静脉是一个优越的位置, 血液取样4。然而, 从大鼠的低血流尾静脉中获得足够的血量并不总是容易的。真空采血系统通常用于从患者的日常临床实践中采集血液样本。该真空采血系统已在患者和研究动物中使用了很长时间8,9,10。该系统由真空采血管与硅胶内管壁和蝶形针组成。它是基于真空负压原理, 并允许连续绘制的血液没有血液收集的风险, 这也减少了污染和溶血的机会11。导管内血液样本量约0.3 毫升。从尾静脉取出针头后, 导管内的血液会流入收集真空管, 由于真空。该方法也适用于用较小的穿刺针采集小于100克的大鼠血液。
这种采血方法有几个优点。该方法只对大鼠造成最小的损伤, 不使用麻醉剂就可以进行采血, 从而避免了应激反应和麻醉剂对血液样本的影响。我们的观察显示, 在手术过程中, 大鼠在塑料抑制器中是安静的, 在这些实验中没有死亡。第二, 重复采血每一天或两日是可能的, 这使得频繁的血液取样, 以足够的数量, 为各种研究目的。最后, 这一方法是容易学习, 很容易掌握的初学者后, 短 (2 小时) 学习期间, 表明类似的成功率和穿刺时间之间的实验和初学者小组在本报告。在前一份报告中, Lee et al.演示了一种类似的方法, 从鼠的侧尾静脉中抽取血液, 从4。我们的方法比 Lee et al.所报告的更简单4中, 我们直接将穿刺针插入尾静脉, 而另一端则直接连接到真空管, 用于收集血液, 无需使用额外的注射器和手动抽吸血液, 无需 "牛奶静脉, 以促进血液流动。用这个封闭的系统收集血液可以减少血液污染的几率。使用的塑料限制持有人也减少了对老鼠的压力, 并促进固定的尾巴。
以下两个步骤对于成功应用此方法至关重要。1) 延长尾部, 避免尾部移动;2) 在°的角度轻轻穿刺, 避免刺穿静脉, 并保持穿刺针在静脉内。
结论: 大鼠采用的真空采血方法安全、可行、易于操作。这种方法能使血液在非麻醉大鼠中获得足够的血液。

大量的实验研究需要大鼠的血液取样。从大鼠采集血液的方法包括心脏穿刺、逆行眶丛、颈静脉、隐静脉、尾血管、颈动脉、腹主动脉和腔静脉。大多数技术 (除了隐静脉和尾血管穿刺) 需要麻醉1,2。大鼠尾部的静脉被例行穿刺以进行血液取样和血管注射。在用温水处理尾部后, 即使在大鼠被麻醉3时, 血管也能很好地扩张并适合操作。
尽管开发了新的和改进的技术获取血液样品在小实验动物2, 它不总是容易获得足够的血液, 当取样从小动物。大鼠尾静脉是血液取样的优越位置4。然而, 尾巴静脉是稀薄的, 因此它有时是坚硬的刺穿, 并且需要很长时间得到足够的血液由于它的低血液流动。血液采集通过真空抽吸管系统是一种典型的方法在日常临床实践中。这种封闭的血液收集系统减少了直接接触血液的风险, 并使得从单一静脉穿刺中抽取多个样本变得更容易5。本报告介绍了一种简单的尾静脉采血方法在非麻醉大鼠使用真空萃取管系统。该方法易于掌握, 可用于大鼠大量血液的多次提取。

<table><tbody><tr><td>a double-pointed needle</td><td>Shanghai Kang Nong medical instrument co., LTD, China</td><td>22G&amp;nbsp;</td><td>(0.7&amp;times;25mm)</td></tr><tr><td>vacuum tubes</td><td>Wuhan Zhi Yuan, medical science and technology co., LTD, China</td><td>2 ml</td><td></td></tr><tr><td>rat restraining holder&amp;nbsp;</td><td>Shanghai Kang Nong medical instrument co., LTD, China</td><td>250g rat hoder model</td><td></td></tr><tr><td>normal salt diet for rats</td><td></td><td></td><td>Rats received a normal salt diet (0.3% NaCl) throughout the study</td></tr><tr><td>SPSS software for statistical analysis</td><td>SPSS Inc,&amp;nbsp; Chicago; USA&amp;nbsp;</td><td>Version 17.0.&amp;nbsp;</td><td></td></tr></tbody></table>

repeated blood collection from tail vein of non-anesthetized rats with a vacuum blood collection system

血液可以从老鼠体内收集到一些取样地点。例如, 尾部静脉是一个优越的位置, 血液取样。然而, 尾静脉是薄的, 所以有时很难刺穿。此外, 尾静脉有低血流, 需要很长的取样时间, 以获得足够的血液。本报告介绍了一种简单的血液取样方法, 即真空采血法, 通常用于从病人那里获取血液样本, 用于非麻醉大鼠。22克蝴蝶针尖插入到一个侧尾静脉约2-3 厘米从尾部的尖角约°, 血液收集在真空收集管通过插入橡胶年底的蝴蝶针进入 vacuum 血收集管。实验结果表明, 实验组成功率为 95%, 初学者组90%。两组的成功率和穿刺时间相似。实验组的取样时间较初学者组明显缩短。结论: 非麻醉大鼠尾静脉序贯采血采血方法是可行的、易学的, 可作为其他常规采血技术的可靠替代品。鼠.

两组的取样持续时间、体质量和采血量 每组10只老鼠 (5 只雄性) 在两周内采集血液两次。两组的平均体质量相似 (272.00 ±9.66 g 和273.70 ±8.87 g)。实验组的取样持续时间较初学者组明显缩短 (2.77 ±0.53 min vs 3.28 ±0.83, p < 0.05) (表 1)。
两组的成功率和穿刺次数 成功率为 95% (19/20) 在实验组和 90% (18/20) 在初学者组 (p > 0.05)。穿刺次数在两组间无显著差异 (1.50 ±0.61 分钟与1.85 ±0.75 分, p > 0.05) (表 2)。结果表明, 该方法易于学习, 在2小时训练后可以掌握该技术。
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/55852/55852fig1.jpg"> 图 1:2 毫升真空采血管 (左); 22 克蝶针 (中心); 塑料限制持有人 (右).<a href="//cloudflare.jove.com/files/ftp_upload/55852/55852fig1large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 2" class="xfigimg" src="/files/ftp_upload/55852/55852fig2.jpg"> 图 2:针尖入到侧尾静脉 ( 大约 5 毫米 ) 在大约 2 - 3 cm 的位置从尾巴的末端在近似°的角度 .<a href="//cloudflare.jove.com/files/ftp_upload/55852/55852fig2large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 3" class="xfigimg" src="/files/ftp_upload/55852/55852fig3.jpg"> 图 3:成功静脉穿刺.<a href="//cloudflare.jove.com/files/ftp_upload/55852/55852fig3large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always"><tbody>
<tr>
<td colspan="5">实验组</td>
			<td></td>
			<td colspan="5">初学者组</td>
		</tr>
<tr>
<td></td>
			<td>性别</td>
			<td>体重 (g)</td>
			<td>收集的血液容量 (毫升)</td>
			<td>取样持续时间 (分钟)</td>
			<td></td>
			<td></td>
			<td>性别</td>
			<td>体重 (g)</td>
			<td>收集的血液容量 (毫升)</td>
			<td>取样持续时间 (分钟)</td>
		</tr>
<tr>
<td>E1-1</td>
			<td>男性</td>
			<td>291</td>
			<td>1。2</td>
			<td>3.12</td>
			<td></td>
			<td>B1-1</td>
			<td>男性</td>
			<td>282</td>
			<td>1。2</td>
			<td>3.18</td>
		</tr>
<tr>
<td>E1-2</td>
			<td>男性</td>
			<td>291</td>
			<td>1。2</td>
			<td>2.73</td>
			<td></td>
			<td>B1-2</td>
			<td>男性</td>
			<td>282</td>
			<td>1。2</td>
			<td>2.18</td>
		</tr>
<tr>
<td>E2-1</td>
			<td>男性</td>
			<td>276</td>
			<td>1。2</td>
			<td>2。3</td>
			<td></td>
			<td>B2-1</td>
			<td>男性</td>
			<td>264</td>
			<td>1。2</td>
			<td>3.73</td>
		</tr>
<tr>
<td>E2-2</td>
			<td>男性</td>
			<td>276</td>
			<td>1。2</td>
			<td>2.37</td>
			<td></td>
			<td>B2-2</td>
			<td>男性</td>
			<td>264</td>
			<td>1。2</td>
			<td>2.47</td>
		</tr>
<tr>
<td>E3-1</td>
			<td>男性</td>
			<td>267</td>
			<td>1。2</td>
			<td>2.73</td>
			<td></td>
			<td>B3-1</td>
			<td>男性</td>
			<td>265</td>
			<td>1。2</td>
			<td>3.52</td>
		</tr>
<tr>
<td>E3-2</td>
			<td>男性</td>
			<td>267</td>
			<td>1。0</td>
			<td>3.88</td>
			<td></td>
			<td>B3-2</td>
			<td>男性</td>
			<td>265</td>
			<td>1。2</td>
			<td>2.53</td>
		</tr>
<tr>
<td>E4-1</td>
			<td>男性</td>
			<td>263</td>
			<td>1。2</td>
			<td>2.41</td>
			<td></td>
			<td>B4-1</td>
			<td>男性</td>
			<td>289</td>
			<td>1。2</td>
			<td>2.32</td>
		</tr>
<tr>
<td>E4-2</td>
			<td>男性</td>
			<td>263</td>
			<td>1。2</td>
			<td>2.52</td>
			<td></td>
			<td>B4-2</td>
			<td>男性</td>
			<td>289</td>
			<td>1。2</td>
			<td>3.62</td>
		</tr>
<tr>
<td>E5-1</td>
			<td>男性</td>
			<td>265</td>
			<td>1。2</td>
			<td>3.61</td>
			<td></td>
			<td>B5-1</td>
			<td>男性</td>
			<td>277</td>
			<td>1。2</td>
			<td>3.48</td>
		</tr>
<tr>
<td>E5-2</td>
			<td>男性</td>
			<td>265</td>
			<td>1。2</td>
			<td>2.42</td>
			<td></td>
			<td>B5-2</td>
			<td>男性</td>
			<td>277</td>
			<td>1。2</td>
			<td>3.12</td>
		</tr>
<tr>
<td>E6-1</td>
			<td>女性</td>
			<td>285</td>
			<td>1。2</td>
			<td>3.33</td>
			<td></td>
			<td>B6-1</td>
			<td>女性</td>
			<td>274</td>
			<td>0。9</td>
			<td>4.87</td>
		</tr>
<tr>
<td>E6-2</td>
			<td>女性</td>
			<td>285</td>
			<td>1。2</td>
			<td>3.03</td>
			<td></td>
			<td>B6-2</td>
			<td>女性</td>
			<td>274</td>
			<td>1。2</td>
			<td>3.37</td>
		</tr>
<tr>
<td>E7-1</td>
			<td>女性</td>
			<td>271</td>
			<td>1。2</td>
			<td>2.82</td>
			<td></td>
			<td>B7-1</td>
			<td>女性</td>
			<td>268</td>
			<td>1。0</td>
			<td>3.93</td>
		</tr>
<tr>
<td>E7-2</td>
			<td>女性</td>
			<td>271</td>
			<td>1。2</td>
			<td>2.42</td>
			<td></td>
			<td>B7-2</td>
			<td>女性</td>
			<td>268</td>
			<td>1。2</td>
			<td>3.82</td>
		</tr>
<tr>
<td>E8-1</td>
			<td>女性</td>
			<td>268</td>
			<td>1。2</td>
			<td>1.93</td>
			<td></td>
			<td>B8-1</td>
			<td>女性</td>
			<td>262</td>
			<td>1。2</td>
			<td>2.07</td>
		</tr>
<tr>
<td>E8-2</td>
			<td>女性</td>
			<td>268</td>
			<td>1。2</td>
			<td>3.05</td>
			<td></td>
			<td>B8-2</td>
			<td>女性</td>
			<td>262</td>
			<td>1。2</td>
			<td>3.67</td>
		</tr>
<tr>
<td>E9-1</td>
			<td>女性</td>
			<td>273</td>
			<td>1。2</td>
			<td>2.52</td>
			<td></td>
			<td>B9-1</td>
			<td>女性</td>
			<td>281</td>
			<td>1。2</td>
			<td>4.38</td>
		</tr>
<tr>
<td>E9-2</td>
			<td>女性</td>
			<td>273</td>
			<td>1。2</td>
			<td>3.17</td>
			<td></td>
			<td>B9-2</td>
			<td>女性</td>
			<td>281</td>
			<td>1。2</td>
			<td>4.57</td>
		</tr>
<tr>
<td>E10-1</td>
			<td>女性</td>
			<td>261</td>
			<td>1。2</td>
			<td>1.82</td>
			<td></td>
			<td>B10-1</td>
			<td>女性</td>
			<td>275</td>
			<td>1。2</td>
			<td>2.53</td>
		</tr>
<tr>
<td>E10-2</td>
			<td>女性</td>
			<td>261</td>
			<td>1。2</td>
			<td>3.25</td>
			<td></td>
			<td>B10-2</td>
			<td>女性</td>
			<td>275</td>
			<td>1。2</td>
			<td>2.28</td>
		</tr>
<tr>
<td>平均</td>
			<td></td>
			<td></td>
			<td></td>
			<td>2.77</td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td>3.28</td>
		</tr>
<tr>
<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
<tr>
<td colspan="11">En-1: 实验组大鼠的序列号首次采血。</td>
		</tr>
<tr>
<td colspan="11">En-2: 实验组大鼠的序列号, 用于第二次采血。</td>
		</tr>
<tr>
<td colspan="11">Bn-1: 在初学者组的第一个血液收集的大鼠的序列号。</td>
		</tr>
<tr>
<td colspan="11">Bn-2: 在初学者组的第二次血液收集的大鼠的序列号。</td>
		</tr>
</tbody></table>表 1: 实验组和初学者组的血液收集结果。
<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always"><tbody>
<tr>
<td></td>
			<td>血液收集时间</td>
			<td>平均穿刺次数</td>
			<td>一次性穿刺</td>
			<td>两次穿刺</td>
			<td>三次穿刺</td>
			<td>失败</td>
			<td>成功率</td>
		</tr>
<tr>
<td>实验组</td>
			<td>20</td>
			<td>1.50 ±0.61</td>
			<td>11</td>
			<td>8</td>
			<td>0</td>
			<td>1</td>
			<td>95%</td>
		</tr>
<tr>
<td>初学者组</td>
			<td>20</td>
			<td>1.85 ±0.75</td>
			<td>7</td>
			<td>9</td>
			<td>2</td>
			<td>2</td>
			<td>90%</td>
		</tr>
<tr>
<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
<tr>
<td colspan="6">一次性穿刺: 首次穿刺后成功采血的大鼠数量</td>
			<td></td>
			<td></td>
		</tr>
<tr>
<td colspan="6">两次穿刺: 二次穿刺后成功采血的大鼠数</td>
			<td></td>
			<td></td>
		</tr>
<tr>
<td colspan="6">三次穿刺: 第三次穿刺后成功采血的大鼠数量</td>
			<td></td>
			<td></td>
		</tr>
<tr>
<td colspan="5">失败: 第三次穿刺后血液收集失败的老鼠数量</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
</tbody></table>表 2: 实验组和初学者组穿刺次数和成功率的比较。

Watch this Scientific Journal Video about Repeated Blood Collection from Tail Vein of Non-Anesthetized Rats with a Vacuum Blood Collection System at JoVE.com

Repeated Blood Collection from Tail Vein of Non-Anesthetized Rats with a Vacuum Blood Collection System

在这里, 我们描述了一个简单的尾静脉采血方法在非麻醉大鼠使用真空萃取管系统。这种方法减少了直接接触血液的风险, 简化了从单一静脉穿刺中抽取多个样本的可能性。

无麻醉大鼠尾静脉血采集系统的研究

Blood can be collected from rats in a number of sampling locations. For instance, the tail vein is a superior location for blood sampling. However, the ...

repeated-blood-collection-from-tail-vein-non-anesthetized-rats-with

Universidad de Cartagena UDEC

Research

JoVE Journal

Medicine

35.5K 次观看.  Jianghan University. 在这里, 我们描述了一个简单的尾静脉采血方法在非麻醉大鼠使用真空萃取管系统。这种方法减少了直接接触血液的风险, 简化了从单一静脉穿刺中抽取多个样本的可能性。

视频: Repeated Blood Collection from Tail Vein of Non-Anesthetized Rats with a Vacuum Blood Collection System

Collection Protocol for Human Pancreas

This dissection and sampling procedure was developed for the Network for Pancreatic Organ Donors with Diabetes (nPOD) program to standardize preparation of pancreas recovered from cadaveric organ donors. The pancreas is divided into 3 main regions (head, body, tail) followed by serial transverse sections throughout the medial to lateral axis. Alternating sections are used for fixed paraffin and fresh frozen blocks and remnant samples are minced for snap frozen sample preparations, either with or without RNAse inhibitors, for DNA, RNA, or protein isolation. The overall goal of the pancreas dissection procedure is to sample the entire pancreas while maintaining anatomical orientation.
Endocrine cell heterogeneity in terms of islet composition, size, and numbers is reported for human islets compared to rodent islets 1. The majority of human islets from the pancreas head, body and tail regions are composed of insulin-containing &beta;-cells followed by lower proportions of glucagon-containing &alpha;-cells and somatostatin-containing &delta;-cells. Pancreatic polypeptide-containing PP cells and ghrelin-containing epsilon cells are also present but in small numbers. In contrast, the uncinate region contains islets that are primarily composed of pancreatic polypeptide-containing PP cells 2. These regional islet variations arise from developmental differences. The pancreas develops from the ventral and dorsal pancreatic buds in the foregut and after rotation of the stomach and duodenum, the ventral lobe moves and fuses with the dorsal 3. The ventral lobe forms the posterior portion of the head including the uncinate process while the dorsal lobe gives rise to the rest of the organ. Regional pancreatic variation is also reported with the tail region having higher islet density compared to other regions and the dorsal lobe-derived components undergoing selective atrophy in type 1 diabetes4,5.
Additional organs and tissues are often recovered from the organ donors and include pancreatic lymph nodes, spleen and non-pancreatic lymph nodes. These samples are recovered with similar formats as for the pancreas with the addition of isolation of cryopreserved cells. When the proximal duodenum is included with the pancreas, duodenal mucosa may be collected for paraffin and frozen blocks and minced snap frozen preparations.

This video demonstrates a dissection procedure for processing human pancreas into multiple storage formats. Anatomical orientation is maintained throughout the pancreatic regions to allow definition of regional islet composition and density.

人体胰腺的收集协议

This dissection and sampling procedure was developed for the Network for Pancreatic Organ Donors with Diabetes (nPOD) program to standardize preparation ...

科研

医学

Modified Blood Collection from Tail Veins of Non-anesthetized Mice with a Vacuum Blood Collection System and Eyeglass Magnifier

Blood sample collection is the basis of experimental animal research. It is of importance to obtain adequate blood samples for various research purposes. The tail veins of mice are small, and it is sometimes difficult to obtain the required blood volume by conventional puncture methods. This study investigates the superiority of repeated blood sample collection from tail veins of mice through use of a vacuum blood collection system and eyeglass magnifier (experimental group) compared to conventional blood sampling methods (conventional group), performed by beginners and experts, respectively. With the help of an eyeglass magnifier, a butterfly needle tip is inserted into the tail vein of each mouse in the experimental group. When the vein is penetrated successfully, a blood sample is collected in the vacuum collection tube by insertion of the rubber end of a butterfly needle into the vacuum blood collection tube. The plunger is then used to collect blood without the help of the eyeglass magnifier in the conventional group. Success rates of blood sample collection by the beginners and experts were shown to be 70% vs. 100% (p &#60; 0.01) in the experimental group and 35% vs. 85% (p &#60; 0.01) in the conventional group. For both beginners and experts, puncture times required for obtaining required blood sample were significantly lower in the experimental group compared to the conventional group (2.40 &#177; 0.75 vs. 2.90 &#177; 0.31, p &#60; 0.05; 1.15 &#177; 0.37 vs. 1.55 &#177; 0.76, p &#60; 0.05). In conclusion, the presented blood sampling technique is feasible and easy to practice and enables frequent sampling of adequate blood volumes from non-anesthetized mice.&#160;

This study reports blood sampling from tail vein in mice using a vacuum extraction tube system with eyeglass magnifier. Our method is easy to practice and could be used for repeat blood sampling in mice.

真空采血系统和眼镜放大镜从非麻醉小鼠尾静脉中修饰采血

Blood sample collection is the basis of experimental animal research. It is of importance to obtain adequate blood samples for various research purposes. ...

行为学

Subclavian Vein Puncture As an Alternative Method of Blood Sample Collection in Rats

Blood collection with enough blood volume is essential in animal experiments. Blood collection from the tail vein of rats is popular and less stressful compared to other more aggressive methods such as retro-orbital plexus sample collection. However, this blood collection method is sometimes limited by an unsatisfactory success rate. Here, we introduce a method for blood collection through the subclavian vein puncture. The subclavian vein is located just under the clavicle and this vein is large enough to fulfill the volume requirements of blood collection. Our results show that this method is safe and applicable for blood collection sampling with the required blood volume. Blood collection through the subclavian vein puncture could serve as an alternative blood collection method in case of failed tail vein blood sampling in rats.

Here, we present a protocol to collect blood samples from the rat subclavian vein.

锁骨下静脉穿刺作为大鼠采血的一种替代方法

Blood collection with enough blood volume is essential in animal experiments. Blood collection from the tail vein of rats is popular and less stressful ...

Sampling Blood from the Lateral Tail Vein of the Rat

Blood samples are commonly obtained in many experimental contexts to measure targets of interest, including hormones, immune factors, growth factors, proteins, and glucose, yet the composition of the blood is dynamically regulated and easily perturbed. One factor that can change the blood composition is the stress response triggered by the sampling procedure, which can contribute to variability in the measures of interest. Here we describe a procedure for blood sampling from the lateral tail vein in the rat. This procedure offers significant advantages over other more commonly used techniques. It permits rapid sampling with minimal pain or invasiveness, without anesthesia or analgesia. Additionally, it can be used to obtain large volume samples (upwards of 1 ml in some rats), and it may be used repeatedly across experimental days. By minimizing the stress response and pain resulting from blood sampling, measures can more accurately reflect the true basal state of the animal, with minimal influence from the sampling procedure itself.

Blood samples are useful for assessing biomarkers of physiological states or disease in vivo. Here we describe the methodology to sample blood from the lateral tail vein in the rat. This method provides rapid samples with minimal pain and invasiveness.

从大鼠的侧尾静脉血液取样

Blood samples are commonly obtained in many experimental contexts to measure targets of interest, including hormones, immune factors, growth factors, ...

生物学

同时记录脑电图的大鼠脑脊液和血液

Sampling Cerebrospinal Fluid and Blood from Lateral Tail Vein in Rats During EEG Recordings

Because the composition of body fluids reflects many physiological and pathological dynamics, biological liquid samples are commonly obtained in many experimental contexts to measure molecules of interest, such as hormones, growth factors, proteins, or small non-coding RNAs. A specific example is the sampling of biological liquids in the research of biomarkers for epilepsy. In these studies, it is desirable to compare the levels of molecules in cerebrospinal fluid (CSF) and in plasma, by withdrawing CSF and plasma in parallel and considering the time distance of the sampling from and to seizures. The combined CSF and plasma sampling, coupled with video-EEG monitoring in epileptic animals, is a promising approach for the validation of putative diagnostic and prognostic biomarkers. Here, a procedure of combined CSF withdrawal from cisterna magna and blood sampling from the lateral tail vein in epileptic rats that are continuously video-EEG monitored is described. This procedure offers significant advantages over other commonly used techniques. It permits rapid sampling with minimal pain or invasiveness, and reduced time of anesthesia. Additionally, it can be used to obtain CSF and plasma samples in both tethered and telemetry EEG recorded rats, and it may be used repeatedly across multiple days of experiment. By minimizing the stress due to sampling by shortening isoflurane anesthesia, measures are expected to reflect more accurately the true levels of investigated molecules in biofluids. Depending on the availability of an appropriate analytical assay, this technique may be used to measure the levels of multiple, different molecules while performing EEG recording at the same time.

作者聚焦:获取高质量的 CSF 和血液样本以发现癫痫生物标志物 

The protocol shows repeated cerebrospinal fluid and blood collections from epileptic rats performed in parallel with continuous video-electroencephalogram (EEG) monitoring. These are instrumental for exploring possible links between changes in various body fluid molecules and seizure activity.

在脑电图记录期间从大鼠的侧尾静脉采集脑脊液和血液

Because the composition of body fluids reflects many physiological and pathological dynamics, biological liquid samples are commonly obtained in many ...

神经科学

Repetitive Blood Sampling from the Subclavian Vein of Conscious Rat

Rats are widely used in pharmacokinetics (PK) and toxicokinetic (TK) studies that need to collect a certain amount of blood at specific time points to detect drug exposure. The rat blood sampling method determines the quality of the plasma and further affects the precision of the test results. The subclavian vein blood collection method described in this protocol collects blood samples repeatedly in the conscious state of animals to meet the needs of PK and TK tests. The skills of restraint handling and appropriate procedure of needle incision ensure the success rate of blood collection. It is easy to operate while ensuring the quality of plasma and at the same time catering to animal welfare. However, this method requires skilled operation, and an improper one may cause animal weakness, pain, lameness, and even mortality. The current method has been used in the testing facility for a 4-week oral toxicity study in Sprague Dawley (SD) rats with TK. The maximum amount of blood collected within 24 h did not exceed 20% of the animal's total blood. The animals' body weight was more than 200 g for males and females. The data showed that the animals' bodyweight increased steadily every week, and the clinical observation was normal after repetitive sample collection.

The present protocol describes a simple and efficient method for collecting blood from the subclavian vein in rats. It enables rapid, timely, and easily identifiable sampling without anesthesia and obtains high-quality blood through repetitive sample collection.

意识大鼠锁骨下静脉的重复血液采样

Rats are widely used in pharmacokinetics (PK) and toxicokinetic (TK) studies that need to collect a certain amount of blood at specific time points to ...

大鼠采血技术的比较研究

Modified Tail Vein and Penile Vein Puncture for Blood Sampling in the Rat Model

Blood samples are required in most experimental animal designs to assess various hematological parameters. This paper presents two procedures for blood collection in rats: the lateral tail vein puncture and the dorsal penile vein puncture, which offer significant advantages over other previously described techniques. This study shows that these two procedures allow for fast sampling (under 10 min) and yield sufficient blood volumes for most assays (202 &#956;L &#177; 67.7 &#956;L). The dorsal penile vein puncture must be done under anesthesia, whereas the lateral tail vein puncture can be done on a conscious, restrained animal.
Alternating these two techniques, therefore, enables blood draw in any situation. While it is always recommended for an operator to be assisted during a procedure to ensure animal welfare, these techniques require only a single operator, unlike most blood sampling methods that require two. Moreover, whereas these previously described methods (e.g., jugular stick, subclavian vein blood draw) require extensive prior training to avoid harm to or death of the animal, tail vein and dorsal penile vein puncture are rarely fatal. For all these reasons, and according to the context (e.g., for studies including male rats, during the perioperative or immediate postoperative period, for animals with thin tail veins), both techniques can be used alternately to enable repeated blood draws.

作者聚焦：大鼠的上下文依赖性采血方法 

Here, we present a protocol to offer rapid, easy, and reliable blood collection alternatives for the rat model. We describe three different blood sampling methods according to the context: tail vein puncture under anesthesia or on a conscious animal, and dorsal penile vein puncture under anesthesia.

改良尾静脉和静脉穿刺用于大鼠模型中的血液采样

Blood samples are required in most experimental animal designs to assess various hematological parameters. This paper presents two procedures for blood ...

无麻醉大鼠尾静脉血采集系统的研究

摘要

探索更多视频

此视频中的章节

人体胰腺的收集协议

真空采血系统和眼镜放大镜从非麻醉小鼠尾静脉中修饰采血

锁骨下静脉穿刺作为大鼠采血的一种替代方法

从大鼠的侧尾静脉血液取样

在脑电图记录期间从大鼠的侧尾静脉采集脑脊液和血液

在脑电图记录期间从大鼠的侧尾静脉采集脑脊液和血液

在脑电图记录期间从大鼠的侧尾静脉采集脑脊液和血液

意识大鼠锁骨下静脉的重复血液采样

改良尾静脉和静脉穿刺用于大鼠模型中的血液采样

改良尾静脉和静脉穿刺用于大鼠模型中的血液采样