By Gareth Griffiths
Electron microscopy within the organic sciences will be divided into disciplines. the 1st, considering excessive solution element of debris or periodic constructions, is usually according to sound theoretical rules of physics. the second one, via some distance the bigger self-discipline, is attracted to the data available from skinny sections. The theoretical again floor to these teams of options for getting ready and looking out at skinny sections is usually inexact and "loose", for wish of a higher be aware. What will be chemistry is frequently in the direction of alchemy. this type of electron microscopy is frequently enshrined with mystical recipes, passed down from new release to new release. Admittedly, a number of the tactics concerned, similar to these required to embed tissue in epoxy resins, contain a number of interconnected steps, which make it tough to stick with the main points of somebody of those steps. If these kinds of steps are shrouded in a few secret, even though, can one quite 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 larger approach to do it than to categorize the parameters with ++, +/-, and so on? 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 influence that a number of the multi-authored textbooks that existed then (and that experience seemed given that) tended to exacerbate or at the very least perpetuate this
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Additional resources for Fine Structure Immunocytochemistry
Thirty-five years later, however, despite tremendous progress in recent years, there is still no comprehensive model of the Golgi complex which can completely explain why it needs a set of flattened cisternae. A number of new structures have "appeared" along with improvement in specimen preparation techniques, especially during the era following the introduction of glutaraldehyde as a fixative (Sabatini et al. 1963). The previous "absence" of such organelles can now, in a sense, be interpreted as artefactual.
In practice, there are techniques that allow this to be carried out, at least for low resolution studies, and there are also guidelines which allow the quality of the preservation to be subsequently assessed. At all stages in the process caution must be exercised, and if doubt exists it makes sense to use as many independent methods of specimen preparation as possible. This conceptual framework for fine-structural evaluation, the essence of which most microscopists would probably accept as being important for conventional fine-structural studies, must not be abandoned when the additional complication of antibody labelling is thrown into the equation.
J Cell Bioi 82: 114-139 Chapter 3 Fixation for Fine Structure Preservation and Immunocytochemistry Before any cell or tissue section can be viewed in the electron microscope it is always necessary to stabilize or "fix" the sample. The ultimate aim of "fixation" is to "freeze" cell and tissue organization in a particular time frame so that every molecule and, ideally, every ion present in that cell or tissue remains in their original location during visualization. The use of "freeze" is appropriate because the only fIxation procedure which could come close to satisfying the above conditions would be rapid cryo-fixation (= physical fixation of native tissue) followed by cryosectioning and observation of the frozen hydrated sections by cryoelectron microscopy (Chang et al.
Fine Structure Immunocytochemistry by Gareth Griffiths