在生物技术以及医学领域中,常常需要对细胞或组织进行破碎处理从而提取所需要的各种物质。细胞或组织破碎的方法包括机械方法、物理方法、化学方法和生物化学方法等,其中机械法主要通过机械切力的作用使组织细胞破碎,常用的装置有组织捣碎机、匀浆器、超声破碎仪等。
组织捣碎机一般用于动物组织、植物肉质种子、柔嫩的叶芽等,其转速可高达 10000rpm/M以上,由于旋转刀片的机械切力很大,制备一些较大分子如核酸则很少使用。 高压匀浆器是由可产生高压的正向排代泵和排出阀组成,排出阀具有狭窄的小孔,细胞浆液通过止逆阀进入泵体内,在高压下迫使其在排出阀的小孔中高速冲出,并射向撞击环上,由于突然减压和高速冲击,使细胞受到高的液相剪切力而破碎,此法适用于许多微生物细胞的破碎。超声破碎仪将电能通过换能器转换为声能,这种能量通过液体介质而变成一个个密集的小气泡,小气泡迅速炸裂产生的能量起到破碎细胞等物质的作用。
皮肤组织是由表皮、真皮和皮下组织三部分组成。其中,表皮的角质层处在最外面,具有韧性,能够耐受一定的摩擦;而真皮内部的细胞很少,主要由植物纤维结缔组织构成,其中有胶原纤维、弹性纤维和网状纤维等,赋予皮肤一定的弹性 和张力。皮肤组织的这些特性决定了上述常用的研磨方式均不适用于皮肤组织的破碎。研钵是实验室中常用的研磨工具,其配有钵杵,通过钵杵与研钵的挤压与磨擦来达到捣磨、研磨、破碎等目的,其虽能够较为有效地破碎皮肤组织,然而需要进行手工操作,费时费力。
大鼠的皮肤比较厚,因此用上面方法完全研磨这些组织是十分具有挑战性的,使用上海净信冷冻研磨仪,成功实现了对大鼠后爪蛋白质和纸质的研磨提取。
实验方法:
1、去大鼠皮肤组织若干。
2、加缓冲液和净信专用皮肤研磨珠。
3、放入净信冷冻研磨仪中。
4、调好时间频率。
5、得到完美破碎溶液。
6、提心提取上清溶液。
从本实验后续实验人员反馈情况来看,用上海净信冷冻研磨仪和研钵对比来看,优势有这样几个。1、研磨后样品无交叉污染。2、样品后续RNA得率很高。3、研磨样品节省了液氮,降低实验室成本。4、研磨时间得到大大提高,以前几个人工作量减少到一个人几分钟完成批量样品,大大提高了效率。
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8孔可粘腔室载玻片
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µ-Dish 35 mm, low
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New type soil drying box
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24孔板
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API ZYM/ API 50CHB/ API 50CHL
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2孔腔室载玻片
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Gradient amplification PCR detection analyzer THG-96G (gradient)
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API 10S
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灵活可植入探针
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