Download Fine Structure Immunocytochemistry by Gareth Griffiths PDF

By Gareth Griffiths

Electron microscopy within the organic sciences could be divided into disciplines. the 1st, excited by excessive solution aspect of debris or periodic buildings, is generally according to sound theoretical ideas of physics. the second one, by way of a ways the bigger self-discipline, is drawn to the knowledge available from skinny sections. The theoretical again­ floor to these teams of concepts for getting ready and searching at skinny sections is usually inexact and "loose", for wish of a higher be aware. What will be chemistry is frequently toward alchemy. this type of electron microscopy is frequently enshrined with mystical recipes, passed down from new release to new release. Admittedly, a few of the methods concerned, equivalent to these required to embed tissue in epoxy resins, contain a number of interconnected steps, which make it tricky to keep on with the main points of somebody of those steps. If these types of steps are shrouded in a few secret, even though, can one relatively belief the ultimate photo that emerges at the EM reveal? once we current the knowledge in a few semi­ quantitative shape is there particularly no higher technique to do it than to categorize the parameters with ++, +/-, and so forth? What occurs whilst one labels the sections with antibodies? Does the full company necess­ arily must be extra of an "art" than a "science"? Upon reflecting on those difficulties in 1981, I had the impact that the various multi-authored textbooks that existed then (and that experience seemed given that) tended to exacerbate or at the least perpetuate this

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According to dogma, formaldehyde either does not cross-link proteins at all or it cross-links them ineffectively compared with glutaraldehyde. The explanation given is that formaldehyde, unlike glutaraldehyde, has only one functional aldehyde group. Yet many examples where the formaldehyde has given better immunocytochemical labelling than glutaraldehyde have, in recent years, led to a rebirth of interest in its use both for light- and electron microscope immunocytochemistry. 1 Formaldehyde Chemistry At room temperature pure, dry formaldehyde is a colorless gas.

It is not always that easy, however. Many mistakes have been made, and are probably still being made, both in interpreting an artefactual image as a real structure and, conversely, in calling what is in fact a true structure an artefact. This problem can best be illustrated by reference to some historical examples. The Golgi complex is recognized as a central organelle in eukaryotic cells with a pivotal role in secretion and membrane biogenesis (Palade 1975). Described by Camillo Golgi in 1898, it was viewed with some scepticism for several decades, culminating in the light microscopical study by Palade and Claude (1949), claiming it to be an artefact of osmium fixation.

In B, the cells were vitrified without fixation. Note the uniform appearance ofribosomal areas in the cytosol and complete absence of meso somes. x47000, bar I J-lm. Courtesy of Dr. Alasdair McDowall (Howard Hughes Institute, Dallas, Texas) and Dr. Jaques Dubochet (EM Institute, University of Lausanne, Switzerland) from their paper in J. Bact. lA,B 27 28 Fixation for Fine Structure Preservation and Immunocytochemistry definitive or even comprehensive treatise on this subject. Studies in vitro and in vivo have indicated parameters affecting the quality of fixation.

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