Name: rodent blood collection methods
Uploaded: 2016-09-14T17:46:02.000Z
Duration: 11 min 51 s
Channel: JoVE

K12 Standards

Next Generation Science Standards

High School

Life Science

from-molecules-to-organisms-structures-and-processes

From Molecules to Organisms: Structures and Processes

HS-LS1-2

Blood Withdrawal I

此处所述的所有动物实验操作必须遵循机构动物伦理指南，并经IACUC批准。所有操作须遵守3R原则——替代（Replacement）、减量（Reduction）和优化（Refinement），并由经过培训的人员执行。
1. 尾静脉采血
<ol>
	<li>设备
	<ol>
		<li>塑料固定架</li>
		<li>带橡胶塞的真空采血管</li>
		<li>双头（蝶形）针头（22G）</li>
		<li>放大眼镜</li>
		<li>浸有70%乙醇的棉球</li>
		<li>温水（20–30°C）</li>
	</ol>
	</li>
	<li>固定
	<ol>
		<li>将小鼠放入塑料固定架中。</li>
		<li>前肢和后肢应舒适放置，以尽量减少应激和移动。</li>
		<li>尾部伸直并保持固定，以便采样。</li>
	</ol>
	</li>
	<li>采血
	<ol>
		<li>使用吸入性麻醉剂（如异氟烷）在钟形罩或气体麻醉诱导室中对动物进行麻醉，直至达到有效麻醉深度。</li>
		<li>动物完全麻醉后，将其取出并置于侧卧位。</li>
		<li>通过将手指置于头部上方并沿下颌线向下施力，向后下方牵拉皮肤，使眼球突出。</li>
		<li>避免对气管施加压力，以免导致气道塌陷或阻塞，引起窒息死亡。</li>
	</ol>
	</li>
	<li>尾静脉采血
	<ol>
		<li>用温水（20–30°C）清洗尾部，以促进血管扩张。切勿使用热水。</li>
		<li>随后用70%乙醇擦拭尾部表面，清洁皮肤并进一步增强静脉可见度。</li>
		<li>选择一条侧尾静脉进行采样。轻轻抓握并固定尾部下段。可使用放大眼镜以改善静脉的可视化效果。</li>
		<li>将22G蝶形针头以约10°的角度插入尾部侧面静脉，进针位置约为距尾尖一半处，针头方向朝向尾根部，以便重复采样。</li>
		<li>将针头的橡胶端插入采血管，利用真空负压收集血液。</li>
		<li>可根据批准的实验方案从尾部获取多个样本。进行微量采血时，使用细针穿刺侧尾静脉，以获得小体积且可控的血液样本。</li>
	</ol>
	</li>
</ol>
2. 尾部剪取
<ol>
	<li>设备
	<ol>
		<li>小动物固定器（已消毒）</li>
		<li>锋利的手术剪刀（或 preferably 无菌手术刀片）</li>
		<li>肝素化毛细管</li>
		<li>吸球</li>
		<li>含钾 EDTA 的微量采血管（已标记）</li>
		<li>异丙醇</li>
		<li>局部麻醉霜</li>
		<li>无菌纱布或棉签（用于止血）</li>
	</ol>
	</li>
	<li>固定
	<ol>
		<li>将小鼠轻柔地放入已消毒的固定器中。</li>
		<li>固定固定器以尽量减少操作过程中小鼠的移动。</li>
		<li>调整尾巴位置，以便于接触尾尖。</li>
	</ol>
	</li>
	<li>采血
	<ol>
		<li>使用新鲜的异丙醇清洁尾巴。根据需要在尾尖涂抹局部麻醉霜。</li>
		<li>使用锋利的剪刀（或 preferably 手术刀片），切除约 0.5–1 mm 的尾尖组织。</li>
		<li>直接将血液收集至肝素化毛细管中（约 80 µL）。</li>
		<li>使用吸球将收集的血液转移至标记好的含钾 EDTA 微量采血管中。轻轻倒置试管约 20 次，以防止凝血。</li>
	</ol>
	</li>
	<li>止血与恢复
	<ol>
		<li>通过使用无菌纱布对尾尖施加轻柔压力，直至出血停止，以控制出血。</li>
		<li>如有需要，可谨慎使用其他方法（如电灼或止血剂）以达到止血效果。</li>
		<li>在将动物放回饲养笼之前和之后，必须对其进行监测，以确保实现止血。​</li>
	</ol>
	</li>
</ol>
3. 心脏采血
<ol>
	<li>设备
	<ol>
		<li>对于小鼠，推荐使用带有22-25号×1英寸针头的3 cc注射器。较小的注射器无法提供相同的回压，可能导致采血更加困难。小于25号的针头会限制血液流动，导致凝血增加并损伤血细胞。短于1英寸的针头从膈肌进针时可能无法到达心脏水平。</li>
		<li>对于大鼠，推荐使用带有18号×1.5英寸针头的10-12 cc注射器。根据大鼠体型的不同，较小的注射器可能无法容纳全部待采集的血量，因此在操作过程中需要更换注射器。小于20号的针头会限制血液流动，导致凝血增加。短于1.5英寸的针头从膈肌进针时可能无法到达心脏水平。</li>
		<li>使用足够大小的采血管来盛装采集的血液。</li>
	</ol>
	</li>
	<li>固定
	<ol>
		<li>正确的固定对该方法的成功至关重要。应抓住动物颈背部皮肤，使身体垂直悬垂。必须保持身体笔直，以防止心脏偏移或胸腔扭曲。</li>
		<li>另一种可选体位是仰卧位，此时将针头插入动物左侧肋骨之间。该体位特别适用于体型非常大的大鼠，或需要对多只动物进行采血的情况。</li>
	</ol>
	</li>
</ol>
<img alt="动物实验、注射方法、实验小鼠、注射器使用、科研操作演示。" src="/files/ftp_upload/10246/10246fig1v3.jpg" data-altupdatedat="1784001604000" data-alt="图1"> 
图1. 小鼠垂直持握状态下的心脏采血。
<ol start="3">
	<li>采血
	<ol>
		<li>从后方入路，穿刺膈肌时，若将小鼠或大鼠用颈背部皮肤垂直抓握，操作更为方便。
		<ol>
			<li>将针头向前推进至动物剑突左侧的切迹处。</li>
			<li>针头应与脊柱平行，并置于肋骨稍下方。</li>
			<li>心脏的位置大约与肘部齐平。</li>
			<li>将针头斜面向上插入胸腔，并穿刺心脏。</li>
			<li>用注射器施加轻微的负压。若针头位于心脏内，血液将流入注射器。</li>
			<li>待血液充满注射器后，再增加额外的负压。</li>
		</ol>
		</li>
		<li>从动物左侧进行侧向入路时，需将动物置于仰卧位。
		<ol>
			<li>穿刺点的定位以胸壁上肘部对应点为参照。心脏的位置大约与肘部齐平。</li>
			<li>针头在胸壁中点处垂直于实验台平面插入，该中点为背腹方向测量所得。</li>
			<li>将针头斜面向上插入胸腔，并穿刺心脏。</li>
			<li>用注射器施加轻微的负压。若针头位于心脏内，血液将流入注射器。</li>
			<li>待血液充满注射器后，再增加额外的负压。</li>
		</ol>
		</li>
	</ol>
	</li>
</ol>
<img alt="小鼠解剖实验；用于采血的注射器；生物学研究的实验操作。" src="/files/ftp_upload/10246/10246fig2v3.jpg" data-altupdatedat="1784001604000" data-alt="图 2"> 
图 2. 小鼠仰卧位时的心脏采血。
<ol start="4">
	<li>技术提示
	<ol>
		<li>正常心脏的位置为心尖朝向左侧。在极少数情况下，心脏可能呈反位，导致穿刺心脏困难。</li>
		<li>注射器上施加过大的回抽压力可能导致心脏塌陷，堵塞针尖斜面，从而阻止血液流入注射器。</li>
		<li>反复施加并释放回抽压力将引发注射器内血液凝固。</li>
		<li>轻柔压迫肝脏可促使更多血量进入循环系统，从而可供抽取。</li>
	</ol>
	</li>
</ol>
4. 眼后静脉丛采血
眶后采血是一种侵入性技术，通常应在适当的麻醉下进行，以最大限度地减少疼痛和痛苦，因为即使由技术熟练的操作者实施，也可能出现明显的不良反应。在许多实验方案中，尤其是当该操作为终点操作时，需要进行深度（终点）麻醉。对于非终点或重复采血，可采用较浅的麻醉而非终点麻醉；然而，此类操作必须经过充分论证，并明确获得机构伦理委员会/动物照护与使用委员会（IACUC）的批准，同时需充分考虑动物福利及累积影响。
<ol>
	<li>设备
	<ol>
		<li>准备一个麻醉诱导室，用于输送麻醉气体（如异氟烷）。若使用钟形罩，必须确保液态麻醉剂不与动物接触，以避免经皮肤吸收。可使用带小孔的平台。</li>
		<li>推荐使用容量为 50–75 微升的微量压积管。采用聚酯薄膜包裹的压积管在操作者手指间更不易破裂，应视为一项安全措施。</li>
		<li>在操作台面上放置数层纸巾或其他隔热材料，以在操作过程中维持动物体温。</li>
	</ol>
	</li>
	<li>动物的准备与体位摆放
	<ol>
		<li>将动物置于钟形罩或气体麻醉诱导室内，使用吸入性麻醉剂（如异氟烷）进行麻醉，直至达到有效麻醉深度。</li>
		<li>动物完全麻醉后，将其取出并置于侧卧位。</li>
		<li>通过将手指置于头部顶部及下颌边缘，向后下方牵拉皮肤，使眼球突出。</li>
		<li>避免对气管施加压力，以防压迫或阻塞气道，导致窒息死亡。</li>
	</ol>
	</li>
	<li>采血
	<ol>
		<li>将微量压积管插入眼内眦，沿鼻部平面以 30–45° 角向尾侧方向插入。</li>
		<li>在轻柔旋转压积管的同时施加压力，以切开结膜膜并破裂眶静脉丛。</li>
		<li>血液将通过毛细作用流入压积管内。</li>
		<li>避免插入过深，以免触及眼窝后部的骨骼。</li>
		<li>一旦开始出血，应持续施压以保持眼球突出状态。</li>
		<li>若需采集多管血液，无需将下一根压积管再次插入眶静脉丛，因血液会持续从内眦流出，可直接收集。</li>
		<li>为停止出血，松开皮肤，使眼球恢复至正常位置。闭合眼睑，并对眼球轻柔加压约 30 秒，以促进止血。</li>
	</ol>
	</li>
</ol>
<img alt="小鼠眼后静脉穿刺、采血方法、实验室实验、研究过程、动物实验。" src="/files/ftp_upload/10246/10246fig3v3.jpg" data-altupdatedat="1784001604000" data-alt="图 3"> 
图 3. 小鼠眼后静脉采血。
关键术语与定义 
在采血操作之前，必须确定所需的样本类型。实验程序可能需要全血、血浆或血清。对于全血样本，必须向样本中添加抗凝剂。血浆是在红细胞分离后仍含有纤维蛋白原和其他凝血因子的液体成分，可从添加了抗凝剂的样本中提取。血清则是通过不加抗凝剂的血液采集获得。当血液形成凝块后，经离心即可得到血清。由于样本已经凝固，血清中不含纤维蛋白原及其他凝血因子。血浆和血清均需使用离心机以2200–2500 RPM转速离心至少15分钟才能获得。 
若需获取全血或血浆样本，必须选用合适的抗凝剂。实验室动物常用的抗凝剂包括肝素、枸橼酸钠和乙二胺四乙酸（EDTA）；具体选择应根据研究需求而定。Sequester——EDTA、肝素和枸橼酸钠的液态形式——可直接注入注射器内壁进行预涂层处理，使抗凝剂在采血过程中立即与血液接触，有助于防止凝血。由于大鼠血液比大多数哺乳动物血液凝固更快，因此在采血时必须使用正确比例的抗凝剂与血液。 
针头的选择取决于动物体型及穿刺部位。通常情况下，针头内径越大，采样速度越快。较大针头的另一个优点是可减少对血细胞的损伤。然而，大内径针头的主要缺点是可能对血管造成损伤。在小鼠和大鼠上，可选用20–29号、长度为0.5–1.5英寸的针头。如果针头过长，不仅使用不便，而且针管内多余的空间可能导致血液凝固。每种采血方法所适用的针头尺寸将在操作步骤部分列出。 
所需样本量也必须预先确定。由于小鼠或大鼠体型较小，进行可存活采血时必须计算最大采血量。一只平均体重为25克的小鼠总血容量约为1.8 ml；一只平均体重为250克的大鼠总血容量约为16 ml。对于未进行液体补充的小鼠或大鼠单次采血，安全可移除的最大血容量为其总血容量的10%，或7.7–8 µl/g。因此，对于一只平均体重的小鼠，其总血容量的10%相当于193–200 µl；对于一只250克的大鼠，则相当于1.9–2.0 ml。研究表明，若移除超过总血容量15%的血液，可能导致低血容量性休克。然而，在进行液体补充的情况下，最多可采集总血容量的15%（即12 µl/g）。对于一只25克的小鼠，相当于300 µl；对于一只250克的大鼠，相当于3 ml。进行液体补充时，液体应预热并皮下给予。 
若需多次采样，则每次采血量应减少。每周最大采血量不得超过总血容量的7.5%，或6 µl/g。对于一只25克的小鼠，相当于每周145–150 µl；对于一只250克的大鼠，相当于每周1.45–1.50 ml。若每两周采样一次，最多可采集总血容量的10%（8 µl/g），即平均体重小鼠每两周200 µl，250克大鼠每两周最多2.00 ml。一项针对平均体重250克大鼠的研究显示，当采血量达总血容量的15–20%时，血液指标恢复正常所需时间超过29天。对于重复采血，液体补充仅能恢复血容量，并不能允许采集更多血液或提高采血频率，因为动物需要时间重新生成血细胞。 
过去，眶后静脉丛采血是一种常见做法。然而，该操作在人道性方面引发了诸多关注。操作过程中，一旦毛细管插入眼内眦后过度移动，可能损伤周围组织，导致眼睑和/或结膜膜肿胀。肿胀组织可能使眼球突出至眼睑无法闭合的程度，进而导致角膜干燥和损伤。肿胀引起的疼痛可能引发动物抓挠和自残行为，最终导致眼球摘除。若在眶后采血过程中毛细管放置不当，可能切断视神经，造成失明。若毛细管推进角度不正确，眼球可能被推出眼眶，导致眼睑滑至眼球后方。一旦发生这种情况，很难将眼球正确复位回眼窝。其他潜在问题还包括脆弱的眼眶骨骨折、眼球穿透导致玻璃体流失，或眼球后方形成血肿，因对眼球及周围结构产生压力而引起剧烈疼痛。尽管存在上述诸多问题，但如果由技术熟练的技术人员操作，并在全身麻醉（如异氟烷吸入麻醉）下充分麻醉动物，眶后采血已被证明是啮齿类动物有效的采血方法。 
小鼠与大鼠的眼眶区域解剖结构有所不同。小鼠眼眶区域存在由血管汇集形成的眶后窦。而大鼠眼眶内虽也有血管丛流经眼球后方，但并不形成类似小鼠那样的窦状结构。因此，该操作在小鼠上更容易实施。通过眶后静脉丛进行重复采样时，两次采血之间至少需间隔10天，以确保局部组织充分愈合。虽然推荐使用全身麻醉，但在小鼠中若事先使用局部眼科麻醉剂（如丙帕卡因或丁卡因），也可在无全身麻醉下完成操作。由于大鼠不具备眶后窦结构，且其眼眶周围膜组织更为坚韧，因此进行该操作时必须实施麻醉。 
通过尾尖剪切采集的样本可能同时含有动脉血和静脉血，并可能混入组织产物污染。若通过揉搓或“挤奶”方式获取更多血液，样本质量会下降。为增加血流量，可用温敷、加热灯或温水浸泡等方式对尾部加热。采血后应对尾尖施加压力以实现止血，并应每隔5–10分钟检查动物，确保已达成止血。重复采样时常出现止血延迟。可使用止血粉辅助止血。首次截尾操作建议使用麻醉（全身或局部），但后续采血通常无需麻醉，尤其是当动物已适应该操作时。麻醉会导致血压下降，从而增加此类采血技术的操作难度。 
许多研究需要通过心脏穿刺放血（致死性采血）获取大量非存活样本。通过心脏穿刺，可从小鼠或大鼠体内采集约一半的总血容量。对于一只平均体重25克的小鼠，相当于40 µl/g或约1 ml血液；一只250克的大鼠则可获得约10 ml血液。放血操作必须在动物麻醉状态下进行。熟练技术人员可使用吸入麻醉，也可使用注射麻醉。但麻醉可能导致血压和循环下降，从而减少采血量。 
心脏穿刺法可在动物麻醉后手动固定进行（闭合法），也可按照尾腔静脉采血法的方案外科暴露心脏后进行（开放法）。闭合法中，针头定位的解剖标志为动物左侧胸廓下缘与剑突交界处形成的凹槽。
本视频将帮助您解答的问题 
小鼠和大鼠的采血可通过多种技术实现。尽管采样量、采样频率以及动物的体型和年龄等多种因素均会产生影响，但最重要的因素是执行采样的技术人员的操作技能。对于本文所述的方法，正确使用麻醉剂对于获得高质量样本以及保障动物福利也至关重要。

血液采集 i

来源： Kay Stewart, RVT, RLATG, CMAR；Valerie A. Schroeder, RVT, RLATG。印第安纳州，圣母大学
血液采集是涉及小鼠和大鼠的研究中常见的操作需求。小鼠和大鼠的采血方法取决于所需血液的体积、采样频率、动物的健康状况以及技术人员的操作熟练程度。本文讨论的所有方法——眶后静脉窦采血、初次尾尖剪切采血和心脏内采血——均需使用全身麻醉。

血液采集 I

来源：Kay Stewart，RVT，RLATG，CMAR；Valerie A. Schroeder，RVT，RLATG。美国印第安纳州圣母大学  
血液采集是……的常见需求

从动物体内采集血液是许多研究项目中的常见需求。在这些动物中选择采血方法时，需考虑多种因素，例如所需血液的体积、采样频率以及待采血动物的健康状况。&#160; &#160; &#160; &#160; &#160; 
本文将回顾这些影响因素，并概述几种血液采集方法，包括尾静脉采血、尾尖剪切采血、心脏穿刺采血以及眶后静脉丛采血。有关其他方法，请参见本系列的第二部视频。&#160; &#160; &#160; 
在深入讨论具体的采血操作规程之前，我们首先回顾一些通用注意事项，包括样本类型、针头选择以及可采集的最大血液体积。&#160; 
在从小鼠或大鼠采集血液前，必须明确实验所需的血液样本类型。实验程序可能需要全血、血浆或血清。 
若采集全血，必须向样本中添加抗凝剂以防止凝固。常用的抗凝剂包括肝素、柠檬酸钠以及乙二胺四乙酸（EDTA）。抗凝剂可直接装入注射器内，以涂布其内表面。&#160; 
这样可在血液进入时立即与抗凝剂接触，有助于防止凝血。由于啮齿类动物血液凝固迅速，必须使用正确的抗凝剂与血液比例。&#160; 
血浆采集需将含抗凝剂的全血进行离心。离心后，位于白细胞和血小板层上方的透明液体即为血浆，其中含有纤维蛋白原及其他凝血因子。&#160; 
而血清则来自不含抗凝剂的全血样本。由于样本已发生凝固，位于上层的血清不含有纤维蛋白原及其他凝血因子。 
针头的选择取决于动物体型及穿刺部位。通常，大口径针头对血细胞造成的损伤较小，且能更快地采集血液，但更易导致血管损伤。 
&#160;针头长度也应予以考虑。若针头过长，操作可能不便，或血液可能在针管内开始凝固。针头选择应基于动物体型和采样部位。&#160; 
最后，由于啮齿类动物体型较小，单次采血所能获取的最大血量存在上限，以避免对机体造成严重伤害。采血时可选择是否进行液体补充——通常使用0.9%生理盐水。&#160; 
各种情况下的上限值列于下方文字版操作规程中。此外，对于需要重复采样的实验，应在每次采血之间提供足够的恢复时间，并考虑进行液体补充，以保障动物福利及血细胞的再生。 
&#160;在回顾了一些通用注意事项后，接下来我们将详细介绍具体的采血技术，首先从尾静脉采血开始。&#160; 
为获取血液样本，先用20至30摄氏度的温水清洗尾部，必要时加热尾部以使侧尾静脉清晰可见，但应使用最低限度的加热。选择右侧或左侧尾静脉进行采血。&#160; 
轻柔地握住尾部下段，并在采血过程中保持尾部伸直。将针尖插入尾尖远端约一半位置处的一条侧尾静脉，进针角度约为10度，方向朝向尾根。&#160; 
通过将针头的橡胶端插入真空采血管，使血液流入真空采血管中。&#160; &#160; &#160; &#160; &#160; &#160; &#160; &#160; &#160; &#160; &#160; &#160; &#160; &#160; &#160; &#160; &#160; &#160; &#160; &#160; &#160; &#160; &#160; &#160; &#160; 
静脉穿刺是另一种允许微量采血的方法。操作时，用针头穿刺尾静脉。一旦进入静脉，使用注射器或毛细管缓慢采集血液，直至达到所需体积。 
尾静脉采血的一种较少使用的替代方法是尾尖剪切法，该方法允许在一段时间内进行多次采血。&#160; 
取一只已麻醉的小鼠，使用锋利的手术剪从尾尖剪去约1至5毫米，并将少量外周血收集至肝素化毛细管中。&#160; 
尾尖剪切法仅应在科学上确有必要、且无更少侵入性替代方法可行的情况下使用，且必须符合机构动物福利指南。采血后应监测动物是否出现任何问题，并提供相应护理，必要时实施人道安乐死。&#160; &#160; &#160; &#160;    
对于需要大量非存活性血液样本的研究，可采用心脏穿刺法。这是一种终末性操作，必须在深度麻醉下进行，并随后实施安乐死，或遵循机构指南执行。&#160; &#160; &#160; &#160; &#160; 
对于心脏穿刺法，应根据物种选择合适的注射器和针头尺寸，以确保高效且安全地完成心脏采血。详情请参见文字说明。 
在实施安乐死后，用一只手抓住啮齿类动物颈背部皮肤，使其身体垂直悬挂。这种固定方式至关重要，因为身体必须保持笔直，以防止心脏偏移或胸腔扭曲。&#160; 
请注意，心脏位置大约与肘部齐平。进针点位于剑突左侧的凹陷处，与脊柱平行并位于肋骨下方。 
将针头斜面朝上插入胸腔并穿刺心脏。用注射器施加轻微回抽压力。若针头位于心脏内，血液将流入注射器。待血液充满针筒后再施加额外的回抽压力。&#160; 
通过心脏穿刺，可从小鼠或大鼠采集约一半的总血量。这相当于从一只普通小鼠采集约1 mL血液，或从一只普通大鼠采集约10 mL血液。 
另一种可选体位是采用侧向进针时的仰卧位。此时，将针头插入动物左侧肋骨之间。进针点应与胸壁上肘部尖端对齐。&#160; 
将针头斜面朝上，垂直于操作台平面，插入胸壁中点位置。用注射器施加轻微回抽压力。若针头位于心脏内，血液将流入注射器。&#160; 
同样，待血液充满针筒后再施加额外回抽压力。请注意，无论何种体位，过大的回抽压力可能导致心脏塌陷，堵塞针头斜面，从而阻止血液流入注射器。 
另一种技术是科学家用于从眼部附近血管采集血液的眶后采血法。请注意，小鼠与大鼠的眼眶区域解剖结构有所不同。&#160; 
大鼠具有位于眼球后方的血管丛，而小鼠则具有形成眶后窦的血管集合，这使得该操作在小鼠中更容易实施。 
首先准备用于采血的毛细管。推荐使用容量为50–75微升的微量血细胞比容管。该操作应在终末麻醉下进行，或遵循机构指南。将已麻醉的动物侧卧放置，呈侧卧体位。 
&#160;接着，用手指按住头部顶部及下颌线，向后下方牵拉皮肤以诱导眼球突出。随后，将微量血细胞比容管插入眼内眦，并以与鼻平面成30至45度角的方向向尾侧推进。&#160; 
在轻轻旋转毛细管的同时施加压力。这将切开结膜膜并破裂眼血管丛或眶后窦。血液将通过毛细作用流入血细胞比容管。一旦血液开始流动，应持续施压以保持眼球突出。&#160; 
为停止出血，松开皮肤，使眼球恢复至正常位置。止血的最佳方法是闭合眼睑并施加轻柔压力约30秒至1分钟。 
由于存在严重眼部损伤和疼痛的风险，眶后采血应被视为最后手段。仅当科学上确有必要、无合适且侵入性更小的替代方法可用、且由经过培训的人员按照机构指南操作时方可使用。&#160; &#160; &#160; &#160; &#160; &#160; &#160; &#160; &#160;    
最后，我们来看这些采血技术的一些应用实例。免疫肿瘤学是一个新兴领域，该领域的研究人员常通过采血来研究癌症发展不同阶段的免疫细胞。 
&#160;此外，干细胞研究人员利用血液样本评估供体细胞在受体系统中整合的成功程度。在此例中，研究人员首先通过尾静脉注射，将雄性小鼠的骨髓细胞移植至野生型和基因修饰的雌性动物体内。&#160; 
随后，他们从受体小鼠的眶后窦采集血液，利用聚合酶链式反应（PCR）分析血细胞的基因组DNA，从而获得供体细胞在两种类型动物体内的嵌合百分比。 
您刚刚观看了JoVE关于采血技术的第一部分内容。请继续观看本系列的下一部视频，了解如何在实验动物中实施其他常用的血液采集技术。&#160;

Education

教育

JoVE Science Education

Basic Biology

基础生物学

Lab Animal Research

实验动物研究

Rodents

啮齿类动物

rodent blood collection methods

Blood collection in mice and rats is a common part of research studies. The method used depends on how much blood is needed, how often samples must be taken, the animal's health, and the technician's skill level.

This topic includes three blood withdrawal methods: retro-orbital sinus bleeds, initial tail snip bleeds, and intracardiac bleeds. Each method requires general anesthesia, which means the animal is made unconscious before the procedure begins.

Rodent Blood Collection Methods

rodent handling and restraint techniques

Minimizing Stress in Mouse and Rat Handling

Mouse and rat handling can create stress, even when the procedure is brief. Cage changing and other noninvasive tasks can raise heart rate, blood pressure, and serum corticosterone levels. These changes may last for several hours. Restraint for injections and blood withdrawal can also affect the animals and may influence scientific data. Because of this, proper training is needed when working with mice and rats. The goal is to reduce the effect of handling on the animals as much as possible. <br...

basic care procedures

Caring for Research Mice and Rats

Research mice and rats need careful housing, feeding, and handling. These animals make up over 90% of the animals used for biomedical research, so their care can strongly affect experiment results. Both immunocompetent animals, which have a normal immune response, and immunocompromised animals need proper support during studies. Some research animals have weakened immune systems and need extra steps to stay healthy. Common examples include naturally occurring immunocompromised mice and mice...

fundamentals of breeding and weaning

Mouse and Rat Breeding Methods

Mouse and rat breeding methods are used in biomedical research to supply animals for laboratory studies. Many research facilities buy mice from commercial breeders, but some also breed mice and rats in-house. In-house breeding can reduce costs and give researchers more options for their studies. Breeding animals in the facility also lets researchers manage genetics and timing. They can plan pregnancies to match research needs and work with embryos and newborns when needed. Mice and rats can be...

rodent identification i

Mice and Rats: Ear Tags and Punch Codes

Proper identification of laboratory mice and rats is important in biomedical research. The right animal must be used for each procedure and for data collection. Clear identification helps keep research records accurate and reliable. This page focuses on permanent identification methods for mice and rats. These methods include ear tags, ear punch codes, microchip implantation, tail tattoos for adult mice, and toe tattoos for neonates. Temporary methods such as dyes and marking pens can also be...

rodent identification ii

Rodent ID Methods for Accurate Records

Accurate rodent identification supports reliable animal records and data collection. Research animal records may include simple cage cards or a detailed database with full information on each animal. Individual identification is the key part of keeping those records correct. Mice and rats can be identified by several methods. These include tattooing, microchip placement, and temporary identification methods. Each method serves a different need in research animal management. The procedure also...

compound administration i

Rodent Injection Routes and Restraint

Rodent injection routes and restraint methods are key parts of compound administration in mice and rats. When a substance must be injected into an animal, the delivery route and the amount of compound must be chosen with care. The right choice helps match the dose to the protocol. Several factors guide the route of administration. These include the pH of the solution, the volume needed for the desired dose, and the viscosity of the solution. Viscosity means how thick or resistant to flow a...

compound administration ii

Choosing the Right Route for Compound Delivery

Compound delivery is a key part of many animal studies. The route used to give the compound can change how it is absorbed. Careful planning helps make sure the compound is delivered correctly. The substance itself also matters when a route is chosen. Its pH, viscosity, and concentration can all affect which method is best. These properties help guide the choice of administration route in a study.

compound administration iii

Specialized Injection Routes in Mice and Rats

Specialized injection routes in mice and rats are sometimes used when standard compound administration methods are not enough. Some study protocols call for less common routes such as intradermal, intranasal, and intracranial injections. These methods are used in laboratory animals and require careful technique. Intradermal injections place a compound into the skin. Intranasal injections deliver a compound through the nose. Intracranial injections are given into the brain. Because these...

compound administration iv

Specialized Injection Routes in Rodents

Specialized injection routes in laboratory mice and rats include intracardiac, footpad, and retro-orbital administration. These methods are used when a protocol calls for a route that is less common than standard compound delivery methods. Each route requires specialized training to perform correctly. Intracardiac injections deliver into the heart, footpad injections target the paw area, and retro-orbital injections are given around the eye region. Because these procedures are more technical,...

blood withdrawal i

Blood collection in mice and rats is a common part of research studies. The method used depends on how much blood is needed, how often samples must be taken, the animal's health, and the technician's skill level. This topic includes three blood withdrawal methods: retro-orbital sinus bleeds, initial tail snip bleeds, and intracardiac bleeds. Each method requires general anesthesia, which means the animal is made unconscious before the procedure begins.

blood withdrawal ii

Rodent Blood Sampling Methods

Blood collection from mice and rats can be done with several methods. The choice depends on the restraint needed, how invasive the procedure is, and whether a general anesthetic is required. These factors help determine which method is best for a given analysis. One older method uses the retro-orbital sinus cavity, which is located behind the eye. This approach has been debated because it may cause tissue damage or even blindness. For that reason, facial vein and submandibular vein bleeding...

anesthesia induction and maintenance

Laboratory Anesthesia and Pain Control

Laboratory anesthesia and pain control are important parts of veterinary care for research animals. The Guide for the Care and Use of Laboratory Animals says that pain assessment and pain relief must be part of that care. Anesthesia means the loss of feeling or sensation.

considerations for rodent surgery 

Aseptic Steps in Rodent Survival Surgery

Aseptic technique is essential for rodent survival surgery. It is used to keep the surgical site as clean as possible and to reduce contamination during the procedure. This approach includes patient preparation, surgeon preparation, and sterilization of instruments and other supplies. It also depends on a clean, controlled environment. Careful planning before surgery helps set up these steps for success. During the operation, animals need close monitoring. After the surgery, postoperative...

diagnostic necropsy and tissue harvest

Rat Necropsy for Tissue Collection

Rat necropsy is a key step for collecting organs and tissues at the end of many animal studies. These final samples are used for data collection time points and later testing. Because the samples are used for analysis, the way they are taken can affect sample quality. Careful collection methods help preserve the organs and tissues for testing. The method of euthanasia, or the procedure used to humanely end the animal's life, can also affect sample quality. This article outlines proper necropsy...

sterile tissue harvest

Aseptic Organ Collection in Rats and Mice

Aseptic organ collection in rats and mice is used to keep harvested tissues free from contamination. The process is important when tissues will be used for cell and tissue cultures. It also supports later protein and RNA analysis, as well as metabolic profiling of tissues. The transcript connects this work to the 3 R's, which were introduced in 1959 by W.M.S. Russell and R.L. Burch in The Principles of Humane Experimental Technique. The 3 R's are replacement, reduction, and refinement in the...

early intervention and pain management

早期干预与疼痛管理

此处所述的所有动物实验操作必须遵循机构动物伦理指南，并经IACUC批准。所有操作均须遵守3R原则——替代（Replacement）、减少（Reduction）和优化（Refinement），且必须由经过培训的人员执行。  监测动物疼痛与痛苦的迹象 识别疼痛的早期征象对于符合伦理的动物研究至关重要。通常通过反映动物整体健康状况的行为、生理和物理参数来对动物进行常规监测。这些参数包括体重变化，持续性体重下降或未能维持预期的体重增长可能提示存在疼痛、疾病或慢性应激。体重评估需结合动物自身的基线数据及预期生长模式，而非仅依据单一数值。其他指标还包括筑巢行为，筑巢不良提示可能存在不适；以及水合状态，通常通过皮肤弹性或黏膜外观进行评估。例如，实验操作后停止理毛、体重下降或忽视筑巢的小鼠可能正处于疼痛状态。活动量减少、摄食或饮水模式改变以及社交互动减少等其他表现也提示动物福利受到损害。 使用小鼠面部表情量表评估疼痛 面部表情评分提供了一种更为敏感的疼痛评估方法。小鼠疼痛面容量表（Grimace...

humane euthanasia and confirmation of death in rodents

啮齿类动物的人道安乐死与死亡确认

此处描述的所有动物实验操作必须遵循机构动物伦理指南，并经IACUC批准。所有操作须遵守3R原则——替代（Replacement）、减少（Reduction）和优化（Refinement），并由经过培训的人员执行。 啮齿类动物的仁慈终点是预先定义的临床、行为或生理指标，用于提示动物正在经历显著的疼痛、痛苦或健康状况下降。当观察到这些体征时，需及时采取干预措施，以最大程度减轻动物痛苦并保障其福利。  化学安乐死 该操作通常通过注射给予过量的批准麻醉剂来开始，具体选择取决于物种和研究方案。注射用麻醉剂（如戊巴比妥钠）可迅速抑制中枢神经系统活性。给药后，动物将失去意识和反射活动。深度麻醉会进一步发展为呼吸抑制，随后心脏和脑功能停止。在诱导前，轻柔操作和适当固定对于最大限度减少动物恐惧和应激至关重要。 吸入剂使用方法 此外，也可使用吸入性麻醉剂实施安乐死。将动物置于受控环境中，通过诱导室或面罩输送麻醉蒸气。诸如异氟烷或七氟烷等试剂可在动物感知疼痛前诱导其失去意识。该方法通过逐步镇静并尽量减少操作，从而降低动物的痛苦。<br...

habituation and positive reinforcement for rodent handling

啮齿类动物操作中的习惯化与正向强化

此处所述的所有动物实验操作必须遵循机构动物伦理指南，并经动物护理和使用委员会（IACUC）批准。所有操作必须遵守“3R”原则——替代（Replacement）、减少（Reduction）和优化（Refinement），并由经过培训的人员执行。 习惯化和正向强化是有助于减少啮齿类动物在常规操作和实验过程中应激的优化策略。这些方法可改善动物福利，促进动物自愿配合，并减少可能影响研究结果的与应激相关的效应。 习惯化 习惯化是一种非联合型学习形式，指动物对重复出现的无威胁性刺激的反应随时间推移而逐渐减弱。通过反复接触操作人员、操作工具或实验程序环境，啮齿类动物会逐渐熟悉这些原本可能引发恐惧或回避行为的经历。习惯化训练应短暂、一致，并逐步引入，以使动物能够按照自身的节奏适应。 正向强化<br...

breeding management in rodent studies

啮齿类动物研究中的繁殖管理

此处描述的所有动物实验操作必须遵循机构动物伦理指南，并经机构动物护理和使用委员会（IACUC）批准。所有操作须遵守“3R”原则——替代（Replacement）、减少（Reduction）和优化（Refinement），并由经过培训的人员执行。 繁殖管理通过减少不必要的动物生产，确保仅产生研究所需的动物数量，从而支持“3R”原则。有效的繁殖策略有助于减少过剩动物，提高动物资源的利用效率，同时保证科学目标的实现。 影响繁殖成功率的因素 育种计划应考虑影响种群繁殖效率的品系特异性及生物学变量，包括交配成功率、每窝产仔数、仔鼠存活率、基因型频率、性别分布、生育力以及繁殖寿命。准确预测这些变量有助于研究人员获得所需数量的实验动物，同时避免过度繁殖。 育种计算<br...

transport and acclimatization of rodents for research

实验用啮齿类动物的运输与适应性驯养

此处描述的所有动物实验操作必须遵循机构动物伦理指南，并经动物护理和使用委员会（IACUC）批准。所有操作须遵守“3R”原则——替代（Replacement）、减少（Reduction）和优化（Refinement），并由经过培训的人员执行。 为科研目的运输啮齿类动物时，需采取相应措施，以尽量减少运输过程中的动物应激并保障动物福利，从而降低对实验结果的干扰。运输笼具应提供充足的通风、垫料、食物以及水合物来源（如凝胶补液包）。小鼠应轻柔操作，放入笼内时避免过度拥挤，以减少应激、攻击行为及受伤风险。笼具应牢固关闭，标明“活体动物”及相关运输信息，并覆盖透气材料，以减少外界刺激，同时确保良好的通风和温度控制。 运输与应激减轻 动物应通过机构指定的路线运输，避免经过公共区域和公共交通系统。公共区域会使动物暴露于过度的噪音、振动、陌生气味、温度波动和人类活动之中，这些因素均可能引发应激反应。运输过程中，应保持笼具直立、水平并固定稳妥，以防止不必要的移动和损伤，从而进一步降低与运输相关的应激。 笼具消毒与动物转移<br...

environmental enrichment for rodents

啮齿类动物的环境富集

此处所述的所有动物实验操作必须遵循机构动物伦理指南，并经IACUC批准。所有操作须遵守3R原则——替代（Replacement）、减少（Reduction）和优化（Refinement），并由经过培训的人员执行。 环境丰容是指采用各种物品和方法来促进实验啮齿类动物的自然行为，改善其心理福祉，并提升动物福利。环境丰容应根据啮齿类动物的物种、品系、年龄、性别及研究设计进行定制，并依照机构的动物护理规程提供。 小鼠的环境富集 筑巢材料与隧道 对于小鼠而言，筑巢材料是最重要的环境丰富化资源之一。垫料和筑巢材料，如木丝、木屑、撕碎的纸张、纸条或巢垫块，均能强烈激发雄性和雌性小鼠的筑巢行为。筑巢行为还可作为健康和福利状况的重要指标，因为处于应激、患病或热应激状态的小鼠可能无法构建充分的巢穴。在笼内添加藏身处，例如隧道，可为小鼠提供安全的躲藏、休息和探索空间。 环境丰富化区域与啃咬物体<br...

rodents

Learn how blood collection is done in mice and rats, and why method choice depends on sample size, timing, animal health, and technician skill.

Rodent Blood Collection Methods | JoVE K12

new-york-state-science-learning-standards

New York State Science Learning Standards

Structure and Function

HS-LS1-2.

HS-LS1-3.

Kay Stewart

Overview

General Considerations for Blood Withdrawal

Tail Vein Bleed

Tail Snip

Cardiac Blood Collection

Retro-Orbital Bleed

Applications

Characteristics of Life

Biology is a natural science that studies life and living organisms, including their structure, function, development, interactions, evolution, ...

https://cloudfront.jove.com/files/media/science-education/science-education-thumbs/10647.jpg

Cell Structure

Background
Cells represent the most basic biological units of all organisms, whether it be simple, single-celled organisms like bacteria, or large, ...

Visualizing Onion and Cheek Cells
ExpandImmediately before the experiment, wash and peel onion bulbs for the class.
Remove the entire brown outer skin and ...

Visualizing Onion and Human Cells
ExpandIn this experiment you will prepare different types of cells from a plant and animal, onion and human ...

What are Cells?

Cell Diversity

The concept of a cell started with microscopic observations of dead cork tissue by Robert Hooke in 1665. Hooke coined the term &#34;cell&#34; based on the ...

Cytoplasm

The cytoplasm consists of organelles and a framework of protein scaffolds called the cytoskeleton suspended in an aqueous solution, the cytosol. The ...

The Nucleus

The nucleus is a membrane-bound organelle that acts as a control center in a eukaryotic cell. It contains chromosomal DNA, which controls gene expression ...

The Central Dogma

The DNA Helix

Nucleic Acids

DNA as a Genetic Template

Structure of a Gene

A gene is the fundamental unit of heredity. Every individual has two copies of each gene, one inherited from each parent. Although most people contain the ...

Chromosome Structure

Base-pairing and DNA Repair

What is Gene Expression?

A gene is a stretch of DNA that serves as the blueprint for functional RNAs and proteins. Since DNA is comprised&#160; of nucleotides and proteins are ...

RNA Structure

The basic structure of RNA consists of a string of ribonucleotides attached by phosphodiester bonds. Although most RNA is single-stranded, it can form ...

Protein Families

Types of RNA

Three main types of RNA are involved in protein synthesis: messenger RNA (mRNA), transfer RNA (tRNA), and ribosomal RNA (rRNA). These RNAs perform diverse ...

Conservation of Protein Domains

Transcription

Transcription is the synthesis of RNA from a DNA sequence by RNA polymerase. It is the first step in producing a protein from a gene sequence. ...

Globular and Fibrous Proteins

Translation

Translation is the process of synthesizing proteins from the genetic information carried by messenger RNA (mRNA). Following transcription, it constitutes ...

Intrinsically Disordered Proteins

Ribosomes

Ribosomes translate genetic information encoded by messenger RNA (mRNA) into proteins. Both prokaryotic and eukaryotic cells have ribosomes. Cells that ...

Protein Complex Assembly

Protein and Protein Structures

Conjugated Proteins

Proteins: From Genes to Degradation

Amyloid Fibrils

What are Proteins?

Proteins are polymers of amino acids linked together by peptide bonds. Proteins and polypeptides are interchangeably used to refer to long chains of amino ...

Ligand Binding Sites

Protein Organization

Proteins are polymers of amino acid residues. They are versatile and responsible for different cellular functions, including DNA replication, molecular ...

Protein-Protein Interfaces

Protein Folding

Proteins are chains of amino acids linked together by peptide bonds. Upon synthesis, a protein folds into a three-dimensional conformation, critical to ...

Conserved Binding Sites

Cofactors and Coenzymes

Enzymes are proteins made of amino acids. The functional group of each constituent amino acid catalyzes a wide variety of chemical reactions via ionic ...

Cooperative Allosteric Transitions

Protein Kinases and Phosphatases

GTPases and their Regulation

Covalently Linked Protein Regulators

protein-complexes-with-interchangeable-parts

Protein Complexes with Interchangeable Parts

Mechanical Protein Function

Structural Protein Function

Protein Networks

Construct an explanation based on evidence for how the structure of DNA determines the structure of proteins, which carry out the essential functions of life through systems of specialized cells. [Assessment Boundary: Assessment does not include identification of specific cell or tissue types, whole body systems, specific protein structures and functions, or the biochemistry of protein synthesis.]

HS-LS1-1

Levels of Organization

Biological organization is the classification of biological structures, ranging from atoms at the bottom of the hierarchy to the Earth&#x27;s biosphere. ...

Animal and Plant Cell Structure

Animal and plant cells not only differ in their structure, function, and mode of nutrition but also in how they reproduce, specialize, and organize into ...

structural-organization-of-the-human-body-an-overview

Structural Organization of the Human Body: An Overview

It is convenient to consider the body&#39;s structures in terms of fundamental levels of organization that increase in complexity: subatomic particles, ...

Overview of Human Digestion

The human body contains a monogastric digestive system. In a monogastric digestive system, the stomach only contains one chamber in which it digests food. ...

Anatomy of the Intestines

Although digestion of proteins, carbohydrates, and lipids may begin in the stomach, it is completed in the intestine. The absorption of nutrients, water, ...

Accessory Organs

Accessory organs are those that participate in the digestion of food but do not come into direct contact with it like the mouth, stomach, or intestine do. ...

Lipid Digestion

Lipids are large molecules that are generally not water-soluble. Since most of the digestive enzymes in the human body are water-based, there are specific ...

Protein Digestion

Protein digestion begins in the stomach, where the highly acidic environment can easily disrupt protein structure by exposing the peptide bonds of ...

Carbohydrate Digestion

Carbohydrate digestion and metabolism break down simple and complex carbohydrates from food into saccharides (i.e., sugars) for the body to use as energy. ...

What is a Nervous System?

Overview

The nervous system is the collection of specialized cells responsible for maintaining an organism&rsquo;s internal environment and coordinating ...

The Parasympathetic Nervous System

Overview

The parasympathetic nervous system is one of the two major divisions of the autonomic nervous system. This parasympathetic system is responsible ...

The Sympathetic Nervous System

Overview

The sympathetic nervous system&mdash;one of the two major divisions of the autonomic nervous system&mdash;is activated in times of stress. It ...

Oogenesis

In human women, oogenesis produces one mature egg cell or ovum for every precursor cell that enters meiosis. This process differs in two unique ways from ...

What is a Sensory System?

Sensory systems detect stimuli&mdash;such as light and sound waves&mdash;and transduce them into neural signals that can be interpreted by the nervous ...

Fertilization

During fertilization, an egg and sperm cell fuse to create a new diploid structure. In humans, the process occurs once the egg has been released from the ...

What is the Skeletal System?

Overview

The adult human skeleton comprises 206 bones that are connected through cartilage, tendons, and ligaments. The skeleton provides a rigid ...

Develop and use a model to illustrate the hierarchical organization of interacting systems that provide specific functions within multicellular organisms. [Clarification Statement: Emphasis is on functions at the organism system level such as nutrient uptake, water delivery, and organism movement in response to neural stimuli. An example of an interacting system could be an artery depending on the proper function of elastic tissue and smooth muscle to regulate and deliver the proper amount of blood within the circulatory system.] [Assessment Boundary: Assessment does not include interactions and functions at the molecular or chemical reaction level.]

Rodent Blood Collection Methods

Description

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