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[噬菌体感染细菌超薄切片的定量研究]

[Quantitative studies on ultrathin sections of bacteria infected with bacterio phage].

作者信息

SECHAUD J, RYTER A, KELLENBERGER E

出版信息

J Biophys Biochem Cytol. 1959 May 25;5(3):469-78.

Abstract

A quantitative relationship has been established between the number of particles, for example bacteriophages, counted in ultrathin sections of bacteria and the total number present in the whole bacterial cells. The factor relating particles counted per section with the total number of these particles per entire bacterium could be arrived at by two methods, which proved to give results in close agreement. The first involves knowledge of the average volume of a bacterial section in proportion to the average volume of a whole bacterium; if the mean number of appearances of the same particle on consecutive sections is also known, may then be calculated. The thickness of sections and, therefore, their volume, as well as the average number of times a single particle is sectioned could be learned by examination of serial sections. By counting the relative number of T2 phage particles which had been intersected once or twice, and relating this proportion to the known phage dimensions, the thickness of the sections was determined to be about 400 A. The second measurement of could be made in a particular case of late phage development where the number of particles per cell was countable or titratable directly in the bacterial lysate, this number being compared with the number seen in sections of the bacteria just before lysis. The different sources of errors are discussed. The statistical error is under 20 per cent, while the systematic errors are higher and cannot yet be indicated precisely. After a very cautious estimation of the upper limits, we can state, however, that the counts made with this method are certainly reliable to well within a factor of two.

摘要

已在细菌超薄切片中计数的颗粒(如噬菌体)数量与整个细菌细胞中存在的颗粒总数之间建立了定量关系。将每切片计数的颗粒与每个细菌中这些颗粒的总数相关联的因子可以通过两种方法得出,事实证明这两种方法得出的结果非常一致。第一种方法需要了解细菌切片的平均体积与整个细菌平均体积的比例;如果还知道同一颗粒在连续切片上出现的平均次数,那么就可以计算出来。通过检查连续切片,可以了解切片的厚度,进而了解其体积,以及单个颗粒被切片的平均次数。通过计数被切片一次或两次的T2噬菌体颗粒的相对数量,并将此比例与已知的噬菌体尺寸相关联,确定切片的厚度约为400埃。在噬菌体发育后期的特定情况下,可以进行的第二次测量,此时每个细胞中的颗粒数量可以在细菌裂解物中直接计数或滴定,将这个数量与裂解前细菌切片中看到的数量进行比较。讨论了不同的误差来源。统计误差在20%以下,而系统误差较高且尚不能精确指出。然而,在非常谨慎地估计了上限之后,我们可以说,用这种方法进行的计数肯定可靠,误差在两倍以内。

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