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 border="0" cellpadding="0" cellspacing="0" width="634">
	<tbody>
		<tr height="71">
			<td height="71" style="height:71px;width:192px;">a double-pointed needle</td>
			<td style="width:244px;">Shanghai Kang Nong medical instrument co., LTD, China</td>
			<td style="width:120px;">22G&#160;</td>
			<td style="width:79px;">(0.7&#215;25mm)</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:192px;">vacuum tubes</td>
			<td style="width:244px;">Wuhan Zhi Yuan, medical science and technology co., LTD, China</td>
			<td style="width:120px;">2 ml</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:192px;">rat restraining holder&#160;</td>
			<td style="width:244px;">Shanghai Kang Nong medical instrument co., LTD, China</td>
			<td style="width:120px;">250g rat hoder model</td>
			<td></td>
		</tr>
		<tr height="140">
			<td height="140" style="height:140px;width:192px;">normal salt diet for rats</td>
			<td></td>
			<td></td>
			<td style="width:79px;">Rats received a normal salt diet (0.3% NaCl) throughout the study</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:192px;">SPSS software for statistical analysis</td>
			<td>SPSS Inc,&#160; Chicago; USA&#160;</td>
			<td style="width:120px;">Version 17.0.&#160;</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

Research

JoVE Journal

Medicine

35.4K 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

Dried Blood Spot Collection of Health Biomarkers to Maximize Participation in Population Studies

Biomarkers are directly-measured biological indicators of disease, health, exposures, or other biological information. In population and social sciences, biomarkers need to be easy to obtain, transport, and analyze. Dried Blood Spots meet this need, and can be collected in the field with high response rates. These elements are particularly important in longitudinal study designs including interventions where attrition is critical to avoid, and high response rates improve the interpretation of results. Dried Blood Spot sample collection is simple, quick, relatively painless, less invasive then venipuncture, and requires minimal field storage requirements (i.e.&#160;samples do not need to be immediately frozen and can be stored for a long period of time in a stable freezer environment before assay). The samples can be analyzed for a variety of different analytes, including cholesterol, C-reactive protein, glycosylated hemoglobin, numerous cytokines, and other analytes, as well as provide genetic material. DBS collection is depicted as employed in several recent studies.

Biomarkers are directly-measured biological indicators of disease or health. In population and social sciences, biomarkers need to be easy to obtain, transport, and analyze. Dried Blood Spot (DBS) collection meets this need, can be collected in the field with high response rates&#160;and analyzed for a variety of biomarkers.

干生生物标志物的血斑收藏最大限度地参与人口研究

Biomarkers are directly-measured biological indicators of disease, health, exposures, or other biological information. In population and social sciences, ...

Collection, Isolation, and Flow Cytometric Analysis of Human Endocervical Samples

Despite the public health importance of mucosal pathogens (including HIV), relatively little is known about mucosal immunity, particularly at the female genital tract (FGT). Because heterosexual transmission now represents the dominant mechanism of HIV transmission, and given the continual spread of sexually transmitted infections (STIs), it is critical to understand the interplay between host and pathogen at the genital mucosa. The substantial gaps in knowledge around FGT immunity are partially due to the difficulty in successfully collecting and processing mucosal samples. In order to facilitate studies with sufficient sample size, collection techniques must be minimally invasive and efficient. To this end, a protocol for the collection of cervical cytobrush samples and subsequent isolation of cervical mononuclear cells (CMC) has been optimized. Using ex vivo flow cytometry-based immunophenotyping, it is possible to accurately and reliably quantify CMC lymphocyte/monocyte population frequencies and phenotypes. This technique can be coupled with the collection of cervical-vaginal lavage (CVL), which contains soluble immune mediators including cytokines, chemokines and anti-proteases, all of which can be used to determine the anti- or pro-inflammatory environment in the vagina.

The use of cytobrush sampling to collect lymphocytes and monocytes from the endocervix is a minimally invasive technique that provides samples for analysis of female genital tract immunity. In this protocol, we describe the collection of cytobrush samples and immune cell isolation for flow cytometry assays.

收集，分离，和人类宫颈样品的流式细胞仪分析

Despite the public health importance of mucosal pathogens (including HIV), relatively little is known about mucosal immunity, particularly at the female ...

Embolic Middle Cerebral Artery Occlusion (MCAO) for Ischemic Stroke with Homologous Blood Clots in Rats

Clinically, thrombolytic therapy with use of recombinant tissue plasminogen activator (tPA) remains the most effective treatment for acute ischemic stroke. However, the use of tPA is limited by its narrow therapeutic window and by increased risk of hemorrhagic transformation. There is an urgent need to develop suitable stroke models to study new thrombolytic agents and strategies for treatment of ischemic stroke. At present, two major types of ischemic stroke models have been developed in rats and mice: intraluminal suture MCAO and embolic MCAO. Although MCAO models via the intraluminal suture technique have been widely used in mechanism-driven stroke research, these suture models do not mimic the clinical situation and are not suitable for thrombolytic studies. Among these models, the embolic MCAO model closely mimics human ischemic stroke and is suitable for preclinical investigation of thrombolytic therapy. This embolic model was first developed in rats by Overgaard et al.1 in 1992 and further characterized by Zhang et al. in 19972. Although embolic MCAO has gained increasing attention, there are technical problems faced by many laboratories. To meet increasing needs for thrombolytic research, we present a highly reproducible model of embolic MCAO in the rat, which can develop a predictable infarct volume within the MCA territory. In brief, a modified PE-50 tube is gently advanced from the external carotid artery (ECA) into the lumen of the internal carotid artery (ICA) until the tip of the catheter reaches the origin of the MCA. Through the catheter, a single homologous blood clot is placed at the origin of the MCA. To identify the success of MCA occlusion, regional cerebral blood flow was monitored, neurological deficits and infarct volumes were measured. The techniques presented in this paper should help investigators to overcome technical problems for establishing this model for stroke research.

Embolic Middle Cerebral Artery Occlusion MCAO for Ischemic Stroke with Homologous Blood Clots in Rats

In this paper, we demonstrate a standard method for producing an embolic middle cerebral artery occlusion with homologous blood clots (fibrin-rich) in adult rat. This model closely mimics human ischemic stroke and is suitable for preclinical study of thrombolytic therapy for ischemic stroke.

栓塞性脑梗塞（缺血）的大鼠缺血性中风与血同源凝块

Clinically, thrombolytic therapy with use of recombinant tissue plasminogen activator (tPA) remains the most effective treatment for acute ischemic ...

Routine Screening Method for Microparticles in Platelet Transfusions

Platelet inventory management based on screening microparticle content in platelet concentrates is a new quality improvement initiative for hospital blood banks. Cells fragment off microparticles (MP) when they are stressed. Blood and blood components may contain cellular fragments from a variety of cells, most notably from activated platelets. When performing their roles as innate immune cells and major players in coagulation and hemostasis, platelets change shape and generate microparticles. With dynamic light scattering (DLS)-based microparticle detection, it is possible to differentiate activated (high microparticle) from non-activated (low microparticle) platelets in transfusions, and optimize the use of this scarce blood product. Previous research suggests that providing non-activated platelets for prophylactic use in hematology-oncology patients could reduce their risk of becoming refractory and improve patient care. The goal of this screening method is to routinely differentiate activated from non-activated platelets. The method described here outlines the steps to be performed for routine platelet inventory management in a hospital blood bank: obtaining a sample from a platelet transfusion, loading the sample into the capillary for DLS measurement, performing the DLS test to identify microparticles, and using the reported microparticle content to identify activated platelets.

Platelet inventory management based on screening microparticle content in platelet concentrates is a new quality improvement initiative in hospital blood banks. The goal is to differentiate activated from non-activated platelets to optimize the use of platelets. Providing non-activated platelets to hematology-oncology patients might reduce their high risk to become refractory.

血小板输血中微粒的常规筛选方法

Platelet inventory management based on screening microparticle content in platelet concentrates is a new quality improvement initiative for hospital blood ...

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 ...

Simultaneous Electrocardiography Recording and Invasive Blood Pressure Measurement in Rats

For studies related to cardiovascular physiology or pathophysiology, blood pressure (BP) and electrocardiography are basic observational parameters. Research focusing on cardiovascular disease models, potential cardiovascular therapeutic targets or pharmaceutical agents requires assessment of systemic arterial pressure and heart rhythm changes. In situations where radio telemetry systems are not available or affordable, the technique of femoral artery cannulation is an alternative way to obtain intra-arterial pressure waveform recordings and systemic BP measurements. This technique is economical and can be performed with standard equipment in animal facilities. However, invasive arterial pressure recording requires cannulation of small arteries, which can be a challenging surgical skill. Here, we present step-by-step protocols for femoral artery cannulation procedures. Key procedures include the calibration of the data acquisition system, tissue dissection and femoral artery cannulation, and setup of the arterial cannulation system for pressure recording. Surface electrocardiography recording procedures are also included. We also present examples of BP recordings from normotensive and hypertensive rats. This protocol allows reliable direct recordings of systemic BP with simultaneous electrocardiography.

Here, we describe a setup for simultaneous recording of electrocardiography and intra-arterial blood pressure (BP) in experimental rats, which can be done with standard equipment in animal facilities and can be applied to physiological or pharmacological studies to investigate pathogenic or therapeutic mechanisms in cardiovascular medicine.

大鼠同时心电图记录及侵入性血压测量

For studies related to cardiovascular physiology or pathophysiology, blood pressure (BP) and electrocardiography are basic observational parameters. ...

A New Best Practice for Validating Tail Vein Injections in Rat with Near-infrared-Labeled Agents

Intravenous (IV) administration of agents into the tail vein of rats can be both difficult and inconsistent. Optimizing tail vein injections is a key part of many experimental procedures where reagents need to be introduced directly into the bloodstream. Unwittingly, the injection can be subcutaneous, possibly altering the scientific outcomes. Utilizing a nanoemulsion-based biological probe with an incorporated near-infrared fluorescent (NIRF) dye, this method offers the capability of imaging a successful tail vein injection in vivo. With the use of a NIRF imager, images are taken before and after the injection of the agent. An acceptable IV injection is then qualitatively or quantitatively determined based on the intensity of the NIRF signal at the site of injection.

Here we present a method to validate tail vein injections in rats by utilizing near-infrared fluorescence imaging data from dyes incorporated into agents or biological probes. The tail is imaged before and after the injection, the fluorescent signal is quantified, and an assessment of the injection quality is made.

近红外标记剂对大鼠尾静脉注射的新的最佳验证

Intravenous (IV) administration of agents into the tail vein of rats can be both difficult and inconsistent. Optimizing tail vein injections is a key part ...

Blood Collection Through Subclavian Vein Puncture in Mice

The mouse is the foremost mammalian model for studying human disease and human health. However, blood sample collection from mice is challenging in research work. Tail blood collection is a popular method when a small amount of blood sample is needed. Orbital artery could be considered if a large amount of blood is needed but this blood collection method has ethical issues. Formerly, we demonstrated the feasibility and safety of blood sample collection through subclavian vein puncture in rats, and here we investigate whether this method could be used in mice. We report that this method is safe and practical for blood collection in mice. Blood collection through the subclavian vein puncture in mice can be a convenient method in daily research works.

Here, we present a protocol to take blood samples from the subclavian vein of mice.

小鼠锁骨下静脉穿刺采血

The mouse is the foremost mammalian model for studying human disease and human health. However, blood sample collection from mice is challenging in ...

Simultaneous Flow Cytometric Characterization of Multiple Cell Types Retrieved from Mouse Brain/Spinal Cord Through Different Homogenization Methods

Recent advances in viral vector and nanomaterial sciences have opened the way for new cutting-edge approaches to investigate or manipulate the central nervous system (CNS). However, further optimization of these technologies would benefit from methods allowing rapid and streamline determination of the extent of CNS and cell-specific targeting upon administration of viral vectors or nanoparticles in the body. Here, we present a protocol that takes advantage of the high throughput and multiplexing capabilities of flow cytometry to allow a straightforward quantification of different cell subtypes isolated from mouse brain or spinal cord, namely microglia/macrophages, lymphocytes, astrocytes, oligodendrocytes, neurons and endothelial cells. We apply this approach to highlight critical differences between two tissue homogenization methods in terms of cell yield, viability and composition. This could instruct the user to choose the best method depending on the cell type(s) of interest and the specific application. This method is not suited for analysis of anatomical distribution, since the tissue is homogenized to generate a single-cell suspension. However, it allows to work with viable cells and it can be combined with cell-sorting, opening the way for several applications that could expand the repertoire of tools in the hands of the neuroscientist, ranging from establishment of primary cultures derived from pure cell populations, to gene-expression analyses and biochemical or functional assays on well-defined cell subtypes in the context of neurodegenerative diseases, upon pharmacological treatment or gene therapy.

We present a flow cytometry method to identify simultaneously different cell types retrieved from mouse brain or spinal cord. This method could be exploited to isolate or characterize pure cell populations in neurodegenerative diseases or to quantify the extent of cell targeting upon in vivo administration of viral vectors or nanoparticles.

通过不同的均质化方法从小鼠大脑/脊髓中检索到的多种细胞类型的同步流细胞测量特征

Recent advances in viral vector and nanomaterial sciences have opened the way for new cutting-edge approaches to investigate or manipulate the central ...