用于流式细胞术计数的大气颗粒物中细菌的分离程序
Detachment Procedure of Bacteria from Atmospheric Particles for Flow-cytometry Counting.
作者信息
Araya Carolina M, Cazorla Alberto, Reche Isabel
机构信息
Departamento de Ecología and Instituto del Agua, Facultad de Ciencias, Universidad de Granada, 18071 Granada, Spain.
Department of Applied Physics, University of Granada, 18071 Granada, Spain.
出版信息
Bio Protoc. 2019 Jun 20;9(12):e3273. doi: 10.21769/BioProtoc.3273.
The protocol separates bacteria from atmospheric particles, obtaining with greater precision their abundance in the atmospheric deposition. This procedure is similar to the one used to separate bacteria in streambed sediments. The detachment procedure consists of a chemical treatment with sodium pyrophosphate and Tween 20 and a physical treatment with agitation and ultrasonic bath to disperse the bacteria in the liquid sample. We recover the total (free and attached) bacteria by generating a density gradient with Nycodenz by centrifugation. The techniques prior to this procedure do not include the microorganisms that are attached to the aerosol particles and, therefore, considerably underestimate the total load and deposition of airborne microorganisms.
该方案将细菌与大气颗粒物分离，从而更精确地获取它们在大气沉降物中的丰度。此程序类似于用于分离河床沉积物中细菌的程序。分离程序包括用焦磷酸钠和吐温20进行化学处理，以及通过搅拌和超声浴进行物理处理，以使细菌分散在液体样品中。我们通过离心用Nycodenz生成密度梯度来回收总细菌（游离和附着的）。在此程序之前的技术不包括附着在气溶胶颗粒上的微生物，因此，大大低估了空气传播微生物的总负荷和沉降量。
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