PolymorphPrep™多型核细胞分离液

PolymorphPrep™分离液(即用型)

订购货号:1114683

规格:250ml

品牌:Axis-Shield

Polymorphprep™ is a ready-made, sterile and endotoxin tested solution used to isolate polymorphonuclear granulocytes from blood. It is the ideal medium for neutrophils and eosinophils. 

The mononuclear and the polymorphonuclear leucocytes are separated into two distinct bands free from red blood cells.

The temperature of the blood sample and the medium should be kept between 18-22°C. 

Temperature variations may affect the density and the viscosity of Polymorphprep(TM) solution which may lead to poor outcome. 

Please note: The method is effective only with whole undiluted blood not with a leukocyte-rich fraction.

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How Polymorphprep™ works

Polymorphonuclear leukocytes (PMNs) has a lower buoyant density than erythrocytes (1.09-1.11 g/ml) but a higher buoyant density than the mononuclear cells (higher than 1.085 g/ml). Therefore, it is difficult to isolate polymorphonuclear granulocytes using a density barrier similar to that used for mononuclear cell. Several procedures have been developed to overcomes these issues. 

The high osmolality of Polymorphprep™ causes erythrocytes to lose water and shrink, thus increasing their effective buoyant densities. This allows the dextran aggregated erythrocytes to sediment rapidly through the dense medium. 

As the cells sediment further into the medium, the osmotic gradient between the medium and the erythrocytes declines. This means that water loss from the erythrocytes is greater at the top of the Polymorphprep(TM) and progressively decreases as they sediment further. A gradient of diatrizoate is formed within the density barrier. The PMNs band within this density gradient while the mononuclear cells remain at the sample/medium interface.

PolymorphPrep™ 是一种即用型、无菌、内毒素含量很低的分离人外周血中多形核粒细胞(polymorphonuclear granulocytes)的分离液。

分离原理:

Polymorphprep™ 分离液的高渗透压可使红细胞失水而缩小,从而增大红细胞的有效浮力密度;分离液中的右旋糖苷能使红细胞聚集并在溶液中快速沉降,随着离心过程红细胞的逐渐沉降,Polymorphprep™ 分离液也形成连续梯度,这样只要离心力和离心时间选择合适,即可将多叶核白细胞停留于分离液中,而红细胞沉降在分离液底部,并且单个核细胞(PBMC)仍就停留于分离液和血液的界面,从而达到同时分离获得多叶核白细胞和单个核细胞的效果。

注意事项:  

  • 该分离液仅适用于人的未作稀释的血液,不适用于动物。

  • 待分离血液和待用分离液的温度需要保持在18度-22度之间,因为温度的变化会影响分离液的密度和粘度,从而影响分离效果。

  • 分离液的保存应置于无菌环境,并避光保存。

相关技术参数:

成分:泛影葡胺钠(Sodium diatrizoate):13.8%(w/v);

右旋糖苷500(Dextran 500):8.0%(w/v)

密度(Density):1.113±0.001g/ml

渗透压(Osmolality):445±15mOsm

内毒素含量(Endotoxins):<1.0EU/ml

分离所得多叶核白细胞中红细胞的含量:整个细胞数目的2-6%。

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