Name: separation of plasmodium falciparum late stage-infected erythrocytes by magnetic means
Uploaded: 2013-03-02T14:00:00
Description: Watch this Scientific Journal Video about Separation of Plasmodium falciparum Late Stage-infected Erythrocytes by Magnetic Means at JoVE.com

这项工作是由授予PRB-009，CS，博士生奖学金，LC，从国家的秘书CIENCIAŸTECNOLOGIA（SENACYT），巴拿马。

所有步骤的协议，除了离心，应进行发动机罩内保持无菌样品。 1。后期分离P.恶性疟原虫感染的红细胞疟原虫感染的红细胞的所有阶段的后期，可以用这种方法，分离自疟原虫色素，赋予对寄生虫的顺磁性，是一种常见的代谢物的属。甲高原虫（3-10％），建议在培养以获得更好的产率与该协议，虽然典型的培养将包含2-4％的血细胞比容（体积的红?...

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在体外培养的疟疾寄生虫P.疟原虫表现出有限的原虫，超过一半的未感染的红血细胞，在最高的增殖培养点。对于大多数的研究实验中，它是理想的工作只与感染的红细胞。为此，一个分离的技术是必要的划分所述培养感染。有用的方法包括使用的链球菌溶血素O，以透性和lyze的未感染RBC 11和一系列利用两组不同密度的变化的差离心。这些方法的一些包括明胶浮选12?...

<table border="1"><tbody><tr><td> 的试剂的名称 </td><td> 公司 </td><td> 目录编号 </td><td> 评论（可选） </td></tr><tr><td> RPMI 1640的Hepes修改</td><td> Sigma-Aldrich公司</td><td> R4130 </td><td>补充有10％人血清，2％葡萄糖和0.2％的碳酸氢钠</td></tr><tr><td> MidiMACS分离</td><td> MACS美天旎生物技术公司</td><td> 130-042-302 </td><td></td></tr><tr><td> MACS MultiStand </td><td> MACS美天旎生物技术公司</td><td> 130-042-303 </td><td></td></tr><tr><td> LS列</td><td> MACS美天旎生物技术公司</td><td> 130-042-401 </td><td></td></tr><tr><td>血球</td><td> Grafco </td><td> Grafco纽鲍尔厅</td><td>可以发现，通过其他许多供应商</td></tr></tbody></table>

separation of plasmodium falciparum late stage-infected erythrocytes by magnetic means

与其他种类的疟原虫，P.恶性疟原虫可以在实验室中培养，这有利于研究1。虽然原虫能够达到≈40％的限制，研究者通常的比例保持在10％左右。在许多情况下，它是必要的隔离含寄生虫的红血细胞（红细胞）来自未感染的，以丰富的文化和继续进行，一个给定的实验。 当P.恶性疟原虫感染的红细胞，寄生虫降解和，馈送来自血红蛋白的2，3。然而，寄生虫必须处理的一个非常有毒的含铁血红素部分4，5。寄生虫逃避其毒性称为haemozoin 6，7成惰性液晶聚合物通过转化的血红素。此含铁分子被存储在其食物泡和在它的金属有一个从一个在血红素8中不同的氧化状态。血红素铁的铁状态ozoin赋予它在未受感染的红细胞没有一个顺磁性。作为入侵的寄生虫成熟，的haemozoin含量也增加9，赋予更顺磁性的 P上的最新的阶段恶性疟原虫红细胞内。 基于此顺磁性，最新的阶段P.恶性疟原虫感染的红血细胞可通过一列含有磁珠通过培养分离。这些珠粒成为磁性的磁体保持器时，被放置在含有它们的列。感染的红细胞，由于其顺磁性，然后将被媚惑​​列，而将包含流过，在大多数情况下，未感染的红细胞和那些含有早期阶段的寄生虫。 在这里，我们描述的方法，以丰富的人口后期寄生虫与的磁性列，保持着良好的寄生虫可行性10。在执行此过程，独立的文化可以返回一个孵化器，让剩余的寄生虫将继续增长。

在图2中，示出的磁性列通过培养之前，（A）和（B）后的程序。一到两个感染的红细胞通常出现在一个放大100倍的字段，如在图2中所示，在图2A中的感染的红细胞上，箭头指向。在一个典型的过程中，在5％的寄生虫血症（ 图2A）与培养开始时，此过程的性能，通常会产生红细胞与原虫的97％-100％（ 图2B）。?...

Watch this Scientific Journal Video about Separation of Plasmodium falciparum Late Stage-infected Erythrocytes by Magnetic Means at JoVE.com

Separation of Plasmodium falciparum Late Stage-infected Erythrocytes by Magnetic Means

顺磁性质的疟原虫色素是用来隔离后期阶段<em&gt;恶性疟原虫</em&gt;感染的红血细胞在培养皿中生长。该方法简单，快速，不影响后续的寄生虫的侵袭能力。

分离<em&gt;恶性疟原虫</em&gt;晚期感染的红细胞由磁性装置

Unlike other Plasmodium species, P. falciparum can be cultured in the lab, which facilitates its study 1. While the parasitemia achieved can reach the ...

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Immunology and Infection

Separation of Plasmodium falciparum Late Stage-infected Erythrocytes by Magnetic Means

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