一种简便的植物单细胞显微操作分选方法

A simple method of sorting plant single cell by micromanipulation

植物细胞具有刚性细胞壁,是植物进行单细胞分析的主要阻碍,选择适当的酶解方法去除细胞壁以后,分选出具有活性的原生质体是进行植物单细胞分析的关键。本文以84 K杨叶肉组织为材料,酶解4 h以后,稀释原生质体至适当密度。在光学显微镜下,使用微吸管、口吸管相互配合,直接进行显微操作分选。移动微吸管针口靠近目标原生质体,缓慢给予负压,将原生质体吸到微吸管内,给予正压释放,成功地分选出目标细胞,分选效率可达100%。该方法可以为多种植物的不同组织单细胞初步分选提供一种简便、快速的方法。

Plant cells have rigid cell walls,which are the main obstacles for the single-cell analysis of plants.After the removal of cell walls by appropriate enzymatic hydrolysis,the separation of active protoplasts is the key to the single-cell analysis of plants.Here,we used 84 K poplar as materials.After 4 hours of enzymatic hydrolysis,we dilute the protoplasts to the appropriate density.Under the optical microscope,the micropipette and mouth pipette are used to cooperate with each other to conduct microscopic separation directly.Move the tip of the micropipette close to the target protoplast,slowly give negative pressure,draw the protoplast into the micropipette,and then give positive pressure to release.As a result,the target cells can be successfully sorted and the sorting efficiency can up to 100%,which provides a simple and fast method for the preliminary separation of plant single cells.