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Sizing Biological Samples by Photosedimentation Techniques

机译:通过光沉降技术对生物样品进行尺寸测定

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AbstractThe performance of the Joyce‐Loebl disk centrifuge in the sizing ofEscherichia colicells, protein inclusion bodies, and cell debris is evaluated. The need for a density gradient that extends throughout the entire spin fluid is highlighted, and a set of standard conditions that fulfill this requirement is defined.E. colicells experience a reduction in their Stokes diameter when exposed to ethanol, indicating that a spin‐buffer fluid combination such as glycerol‐water is to be preferred for the sizing of bacteria. The instrument baseline is influenced by the presence of particles, and a method of estimating the baseline is described. The sizing of small particles is further complicated by baseline drift due to temperature sensitivity of the optical yoke. An analysis of diffusion in the spin fluid is conducted, and an expression for the sedimentation:diffusive flux ratio is derived. For the current samples, it is shown that diffusion within the spin fluid does not lead to significant errors for 0.15‐μm particles, whereas the phenomenon may be significant at the manufacturer's size limit of
机译:摘要评价Joyce-Loebl圆盘离心机对大肠杆菌、蛋白包涵体和细胞碎片进行大小测定的性能。强调了对延伸到整个自旋流体的密度梯度的需求,并定义了一组满足此要求的标准条件.E. 大肠杆菌在暴露于乙醇时其斯托克斯直径减小,表明自旋缓冲液组合(如甘油-水)是细菌大小的首选。仪器基线受颗粒存在的影响,并描述了估计基线的方法。由于光学轭的温度敏感性,基线漂移使小颗粒的尺寸进一步复杂化。对自旋流体中的扩散进行了分析,并推导了沉降:扩散通量比的表达式。对于当前的样品,表明自旋流体内的扩散不会导致 0.15 μm 颗粒的显着误差,而在制造商的尺寸限制下,这种现象可能很显着

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