Some aspects of fixation with glutaraldehyde. A biochemical and histochemical comparison of the effects of formaldehyde and glutaraldehyde fixation on various enzymes and glycogen, with a note on penetration of glutaraldehyde into liver.
Some aspects of fixation with glutaraldehyde. A biochemical and histochemical comparison of the effects of formaldehyde and glutaraldehyde fixation on various enzymes and glycogen, with a note on penetration of glutaraldehyde into liver.
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作者：
D. Hopwood
戊二醛是由Sabatini， Bensch和Barrnett（1963）引入的，作为电子显微镜技术的固定剂。虽然这些工作者对戊二醛对各种酶的作用做了一些粗略的定量估计，但直到最近似乎还没有做进一步的细胞学工作。事实上，制革行业先前对戊二醛最感兴趣，他们已经证明戊二醛是一种强大的交联物质。戊二醛迅速使蛋白质变性的能力也被利用作为消毒剂，它比甲醛或各种苯酚衍生物更有效（Snyder & Cheatle, 1965）。最近，醛类固定剂与金属固定剂的比较已经发表（Ericsson, Saladino & Trump, 1965; Torack, 1965 a; Trump & Ericsson, 1965）。研究发现，醛类物质比四氧化锇更能固定蛋白质。缓冲液的性质、浓度和pH值似乎对含金属固定液比醛类固定液更重要，以获得良好的电子显微照片。Janigan（1964,1965）系统地研究了各种醛对组织磷酸酶和糖苷酶的影响。本工作的目的是扩大戊二醛固定的基本知识。物质的固定性质通常由贝克（1960）概述的各种简单测量来评估。这些指标包括对大鼠肝块的渗透率以及肝块和子宫托的体积变化。由于戊二醛在组织化学和电镜中都有应用，我们从生物化学和组织化学两方面研究了戊二醛对某些酶的影响。在相同的条件下，这些效果与甲醛的效果进行了比较。此外，由于戊二醛将希夫阳性作用于固定在戊二醛中的组织，这一现象得到了进一步的研究。这项工作的初步报告已经出现（Hopwood, 1965）。
Glutaraldehyde was introduced as a fixative for electron microscopic techniques by Sabatini, Bensch & Barrnett (1963). Although these workers did some rough quantitative estimates of the action of glutaraldehyde on various enzymes, little further cytological work seems to have been done until recently. In fact most interest has been shown previously by the tanning industry, who have demonstrated that glutaraldehyde is a powerful cross-linking substance. The ability of glutaraldehyde to rapidly denature protein has also been exploited by using it as a disinfectant, where it is more efficient than formaldehyde or various phenol derivatives (Snyder & Cheatle, 1965). Recently, comparisons ofaldehyde fixatives with metal fixatives have been published (Ericsson, Saladino & Trump, 1965; Torack, 1965 a; Trump & Ericsson, 1965). It was found that aldehydes fix proteins better than osmium tetroxide. The nature, concentration and pH of the buffer seem more important with metal-containing fixatives than with the aldehydes for obtaining good electron micrographs. The effects of various aldehydes on tissue phosphatases and glycosidases have been systematically studied by Janigan (1964, 1965). The aim of the present work is to extend the basic knowledge of glutaraldehyde fixation. The fixative properties of substances are usually assessed by various simple measurements as outlined by Baker (1960). These include rate of penetration into rat liver blocks and volume changes in pessaries and blocks of liver. As glutaraldehyde is used both in histochemistry and electron microscopy its effect on certain enzymes was studied, both biochemically and histochemically. These effects, under similar conditions, were compared with those of formaldehyde. Further, as it was shown, not unexpectedly, that glutaraldehyde introduced Schiff positivity to tissues fixed in it, this phenomenon was further investigated. A preliminary report of this work has already appeared (Hopwood, 1965).