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BD FACSCalibur Flow Cytometry System with 488 nm laser

公司名称: BD
产品编号: FACSCalibur
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Quantification of Bacterial Polyhydroxybutyrate Content by Flow Cytometry
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2017-12-05
[Abstract]  We describe here a detailed protocol for the quantification of the intracellular content of polyhydroxybutyrate (PHB) in a population of bacterial cells by flow cytometry, which is based on a consensus of several previously reported works. [摘要]  我们在这里描述了通过流式细胞术定量细菌细胞群中的聚羟基丁酸酯(PHB)的细胞内含量的详细方案,其基于几个先前报道的作品的共识。

【背景】在营养物质耗尽和存在可利用的碳源的情况下,可以使用几种细菌(例如,Cupriavidus necator,中华根瘤菌Sinorhizobium meliloti, >假单胞菌(Pseudomonas)开启了聚羟基丁酸酯(PHB)中性聚合物的积累,用于储存碳和还原能力(Jendrossek和Pfeiffer,2014)。定量细菌PHB含量的经典方法基于其初始解聚,随后通过紫外分光光度法或气相色谱定量检测β-羟基丁酸的特定化学衍生物(Law and Slepecky,1961; Braunegg等人, 1978)。最近,荧光染料尼罗红(9-二乙氨基-5H-苯并[α]吩恶嗪-5-酮)(Greenspan和Fowler,1985)将PHB颗粒染色(Müller等人,1993)导致了开发新的快速荧光方法来定量细菌样品中聚合物的含量。随后,将基于尼罗红的PHB染色与流式细胞术偶联使得可以定量细菌群体内单细胞水平的聚合物含量(Gorenflo等人,1999; Kacmar等人2006; Tyo等人,2006; Alves等人,2017; Lagares Jr.等人,, ...

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