Name: instillation and fixation methods useful in mouse lung cancer research
Uploaded: 2015-08-31T14:00:00
Description: Watch this Scientific Journal Video about Instillation and Fixation Methods Useful in Mouse Lung Cancer Research at JoVE.com

下面的协议说明，在20-35克鼠效果很好的系统。该方法可以很容易地适应于更大或更小的小鼠通过简单地改变导管的大小。所有动物方案都经约翰霍普金斯大学动物护理和使用委员会。 1.肺灌注<ol><li>选择市售一英寸长的20个克静脉插管以用于插管。 </li><li>修改导管尖端手动弯曲它产生如图1在前端具有微小曲率。 </li><li>麻醉小鼠用氯胺酮（100毫克/千克）和赛拉...

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此处描述的方法有若干优点。第一所需设备是简单且廉价的。第二，该插管可迅速用很少的错误进行。第三，要修复肺以恒定压力，然后测量所述固定肺容积的能力允许结构或细胞在肺10正确定量。 一个可能的缺点插管是小手术。这可能会限制重复该过程如果需要2次滴注的能力。然而，随着粘接剂的精心手术和申请中，我们已经能够每周定期做第二次滴注或?...

由于一些原因，肺癌还没有被广泛研究，在小鼠。其中一个原因是，获得的肺是非常困难的体内 ，并固定肺部定量分析操作并不常见。本文介绍的方法旨在改变这种状况。这里的目标是描述简单的方法，将在设置和小鼠肺分析肺癌或其它病理极大地帮助。而所有这些方法是全新的，它们没有被一起呈现如在这里所描述的简化方式的独立的方法。 已经有许多已描述的方法进行小鼠肺插管主要做重复肺功能或支气管肺泡灌洗在纵向研究单个老鼠的目的手稿。由于该原始文件，但已在该所描述的不同的方法来谅解备忘录几个其他文件Ë插管1-9。虽然所有这些方法都可以成功地被使用，它们通常需要相当大的训练，并且通常不是没有一个非平凡的故障率。另外，在为了进行肺功能测量中，套管需要适应气管得足够紧以便没有空气泄漏。然而，另一个实际使用气管插管是直接提供特定的药物（如癌细胞或其他侮辱），或治疗药物到肺。这样的程序并不需要紧密配合插管，也没有任何复杂的肺功能设备。此处示出了该方法的新颖特征包括一个小的外科手术，其允许插管而不进入食道插管的任何可能性。这个简单的方法使插管成功相对较少的培训或经验。多达30只小鼠/小时可以使用此方法具有故障率接近零对待。 一旦吨他老鼠是准备来牺牲，受伤或癌肺然后可以进行组织学和病理分析中删除。然而，为了正确地量化任何组织学变量与其它的肺相比，有必要标准化的固定程序和适当地量化固定肺容积10。本文详细介绍了简单的程序，使标准化的固定程序，以及一个方法来衡量固定的肺容积。该卷在组织学的量化的重要指标，因为如果没有这样的容积测定，只有相对密度可以测量10。一旦肺容积是已知的，然而，细胞和肺中的其他结构的测量绝对测量然后可以定量。

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>Laboratory Balance</td>
			<td>Ohaus</td>
			<td>Adventurer Pro Model AV 313&#160;</td>
			<td>Other balances can be used if they have a range of 1 - 300g</td>
		</tr>
		<tr>
			<td>20 g Luer intravenous catheter</td>
			<td>Insylte</td>
			<td></td>
			<td>Several other possible vendors, e.g., Jelco Optiva</td>
		</tr>
		<tr>
			<td>500 mL laboratory bottle</td>
			<td>Various</td>
			<td></td>
			<td>Several other possible vendors</td>
		</tr>
	</tbody>
</table>

instillation and fixation methods useful in mouse lung cancer research

灌输活剂，细胞或化学物质直接进入肺而不伤害或杀死小鼠的能力是在肺癌的研究的一个重要工具。虽然有许多方法，这些方法已经发表，显示如何插管老鼠肺功能的测量，没有一个是不为小鼠大同伙快速气管滴注的潜在问题。在本文中，一个简单快捷的方法进行说明，使调查员进行这种滴注以有效的方式。该方法不需要任何特殊的工具或照明，并且可以用很少的实施而得知。它涉及麻醉小鼠，使一个小切口，在颈部以可视化的气管，然后直接插入静脉导管。小切口被快速关闭用组织粘合剂，并且将小鼠允许恢复。一个熟练的学生或技术人员可以在2分钟/小鼠的平均速率做滴注。一旦吨他癌症成立时，经常有必要在肺部的定量组织学分析。传统病理学家平时也懒得固定在规范肺胀，并分析往往是基于一个计分制度，可以说是相当主观的。虽然这有时可能是足够足够的肺肿瘤的大小，总的估计，肺结构或细胞的任何适当的体视学定量分析需要一个可重复的固定程序和后续的肺容积测量。在这里，我们描述了两个固定在压力下的肺部，然后精确测量固定肺容积简单可靠的方法。唯一的要求是一个实验室的平衡，是准确的范围内为1mg-300克。这里提出这样的程序可以大大提高创建，处理和分析肺癌小鼠的能力。

该过程描述在第一协议本身不导致任何广义的结果。这里只描述一个非常可靠的手段来灌输物质直接进入气管。 图7示出了其中锥虫蓝滴注用这里描述的方法的肺的一个例子。有染料的广泛分布，类似于已经看到的直接给出进入气管或小鼠11,12其他染料或示踪剂。我们也用这种方法来提供或者博莱霉素或弹性蛋白酶对肺，这导致广泛纤维化或肺气肿，分别。 <p class="jove_conten...

Watch this Scientific Journal Video about Instillation and Fixation Methods Useful in Mouse Lung Cancer Research at JoVE.com

Instillation and Fixation Methods Useful in Mouse Lung Cancer Research

本文的目的是描述简单的方法，将在设置和小鼠肺分析肺癌或其它病理极大地帮助。我们提出3个协议能够简单，可靠地进行肺滴注，固定，和肺容积的测量。

灌输和固定方法有用的小鼠肺癌研究

The ability to instill live agents, cells, or chemicals directly into the lung without injuring or killing the mice is an important tool in lung cancer ...

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