Mechanisms of Action and Proper Use of Common Fixatives

Dapson, Richard W.

doi:10.1002/9780470875612.ch12

Cited by 5 publications

(4 citation statements)

References 39 publications

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“…142,231 Changes in the tertiary structure may not be the direct result of formaldehyde fixation but rather the subsequent interaction of cross-linked proteins with ethanol or clearing agents during tissue processing. 77,108,261 More energy (eg, antigen retrieval) is needed to reverse formalin fixation cross-links after processing tissues in ethanol, confirming that antigen immunoreactivity is affected not only by formalin fixation but also by postfixation procedures. 108…”

Section: Preanalytical Phase Of Ihcmentioning

confidence: 87%

“…Fixation with formaldehyde is a 3-step process of penetration, covalent bonding, and cross-linking. 51,77 Formation of cross-links between target peptides and irrelevant proteins reduces or blocks their immunoreactivity. 39 Whereas these steps happen simultaneously, they do so at different rates, with penetration being about 12 times faster than bonding and the latter 4 times faster than cross-linking.…”

Section: Preanalytical Phase Of Ihcmentioning

confidence: 99%

“…89,113,129 There is also evidence for a cumulative effect of fixation and tissue-processing factors in the failure of antigen recognition in FFPE from a study of Ki67 and proliferating cell nuclear antigen (PCNA) immunoreactivity in formaldehyde-fixed tissues processed with alcohol and xylene. 77,264 Shifts in the tertiary structure of proteins so that hydrophobic areas are oriented outward and hydrophilic regions inward (the so-called hydrophobic inversion) 77 during dehydration and clearing steps of tissue processing can reduce or abolish antibody binding without antigen retrieval, especially with poorly stabilized (unfixed or suboptimally fixed in formalin) tissues/proteins exposed to a weakly polar or nonpolar solvent. 264 This negative effect varies with the dehydrating and clearing agent used.…”

Section: Preanalytical Phase Of Ihcmentioning

confidence: 99%

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When Tissue Antigens and Antibodies Get Along

2013

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Once focused mainly on the characterization of neoplasms, immunohistochemistry (IHC) today is used in the investigation of a broad range of disease processes with applications in diagnosis, prognostication, therapeutic decisions to tailor treatment to an individual patient, and investigations into the pathogenesis of disease. This review addresses the technical aspects of immunohistochemistry (and, to a lesser extent, immunocytochemistry) with attention to the antigen-antibody reaction, optimal fixation techniques, tissue processing considerations, antigen retrieval methods, detection systems, selection and use of an autostainer, standardization and validation of IHC tests, preparation of proper tissue and reagent controls, tissue microarrays and other high-throughput systems, quality assurance/quality control measures, interpretation of the IHC reaction, and reporting of results. It is now more important than ever, with these sophisticated applications, to standardize the entire IHC process from tissue collection through interpretation and reporting to minimize variability among laboratories and to facilitate quantification and interlaboratory comparison of IHC results.

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Section: Preanalytical Phase Of Ihcmentioning

confidence: 87%

Section: Preanalytical Phase Of Ihcmentioning

confidence: 99%

Section: Preanalytical Phase Of Ihcmentioning

confidence: 99%

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When Tissue Antigens and Antibodies Get Along

2013

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“…The formaldehyde binding can occur with any group containing a reactive hydrogen atom, but the rate varies considerably with amine reactivity being fastest. Cross‐linking then occurs progressively with potential functional groups forming methylene bridges in a process that can continue for months or years (Dapson, 2010). Thus, the increase in fluorescence intensity of L. salmonis samples with storage time can be explained by the slow process of cross‐linking.…”

Section: Discussionmentioning

confidence: 99%

Illuminating the planktonic stages of salmon lice: A unique fluorescence signal for rapid identification of a rare copepod in zooplankton assemblages

Thompson

Bron

Bui

et al. 2021

Journal of Fish Diseases

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Monitoring of planktonic salmon louse (Lepeophtheirus salmonis salmonis) abundance and parameterization of key life‐history traits has been hindered by labour‐intensive and error‐prone quantification using traditional light microscopy. Fluorescence illumination has been proposed as a means of improving visualization, but prior to this study adequate investigation of the relevant fluorescence profiles and measurement conditions has not been undertaken. We investigated the fluorescence profiles of L. salmonis and non‐target copepod spp. with excitation and emission matrices (200–600 nm) and identified unique fluorescence signals. Fluorescence microscopy using excitation wavelengths of 470 ± 40 nm, and emission wavelengths of 525 ± 50 nm, showed that after 90 days of formalin storage salmon lice have a mean fluorescence intensity that is 2.4 times greater than non‐target copepods (copepodid and adult stages). A 7‐day heat treatment of 42°C in formalin increased the difference between salmon louse copepodids and non‐target copepods to a factor of 3.6, eliminating the need for prolonged storage. Differences in the fluorescence signal and endogenous fluorophores were investigated with respect to variation in sea lice species, age, stage and host fish origin. Under the conditions outlined in this paper, the fluorescence signal was found to be a reliable means of visualizing and differentiating salmon lice from non‐target zooplankters. Adaptation of the fluorescence signal would greatly expedite traditional methods of enumerating salmon louse larvae in plankton samples and could provide a means of automated detection.

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Immunohistochemistry

Ramos‐Vara

Borst

2016

Tumors in Domestic Animals

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Mechanisms of Action and Proper Use of Common Fixatives

Cited by 5 publications

References 39 publications

When Tissue Antigens and Antibodies Get Along

When Tissue Antigens and Antibodies Get Along

Illuminating the planktonic stages of salmon lice: A unique fluorescence signal for rapid identification of a rare copepod in zooplankton assemblages

Immunohistochemistry

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