Influence of decalcifying agents on immunoreactivity of formalin-fixed, paraffin-embedded tissue

Matthews, J. B.; Mason, Gillian I.

doi:10.1007/bf01095281

Cited by 49 publications

(28 citation statements)

References 16 publications

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“…7,9,96,232 In a study of melanocytic markers in canine tissues, decalcification for less than a week with formic acid did not significantly reduce immunoreactivity for Melan A, PNL2, or tyrosinase, whereas the use of HCl in the decalcifying solution reduced the immunoreactivity for Melan A and PNL2 after only 1 day of treatment with complete loss of immunoreactivity after 1 week. 295 In that study, tyrosinase was more resistant than the other 2 markers to the deleterious effects of HCl decalcification.…”

Section: Decalcificationmentioning

confidence: 99%

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: Decalcificationmentioning

confidence: 99%

When Tissue Antigens and Antibodies Get Along

2013

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“…Strong acids may affect the detection of some antigens at various degrees. 2,4,35 It is recommended to use weak acidic decalcifying solutions (e.g., formic acid) diluted in formalin. a) Primary antibody characteristics.-There are many characteristics of the antibody that affect incubation time.…”

Section: Practical Approach To Standardization Of a New Antibodymentioning

confidence: 99%

Suggested Guidelines for Immunohistochemical Techniques in Veterinary Diagnostic Laboratories

et al. 2008

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Abstract. This document is the consensus of the American Association of Veterinary Laboratory Diagnosticians (AAVLD) Subcommittee on Standardization of Immunohistochemistry on a set of guidelines for immunohistochemistry (IHC) testing in veterinary laboratories. Immunohistochemistry is a powerful ancillary methodology frequently used in many veterinary laboratories for both diagnostic and research purposes. However, neither standardization nor validation of IHC tests has been completely achieved in veterinary medicine. This document addresses both issues. Topics covered include antibody selection, fixation, antigen retrieval, antibody incubation, antibody dilutions, tissue and reagent controls, buffers, and detection systems. The validation of an IHC test is addressed for both infectious diseases and neoplastic processes. In addition, storage and handling of IHC reagents, interpretation, quality control and assurance, and troubleshooting are also discussed. Proper standardization and validation of IHC will improve the quality of diagnostics in veterinary laboratories.

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“…Although, Matthews et al reported that tissue treated with decalcifying agents may exhibit variable immunoreactivity and impaired counterstaining especially with the use of strong mineralic acids [8]. Overall, laboratories must figure out which type of decalcifying agent to use with the type of bone specimens received, accounting for the amount of mineralized tissue present, which would best ensure an accurate diagnosis.…”

Section: Discussionmentioning

confidence: 99%

Pathology of Metastatic Tumors to Bone: Effects of Decalcification as Experienced at a Single Cancer Center

Henderson-Jackson¹

2016

Oncology and cancer case reports

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IntroductionBone metastases are a frequent complication of renal cancer [1]. Breast and prostate cancer are the most common carcinomas to develop bone metastases, with an incidence of 65-75% and 68%, respectively [2]. In addition, carcinomas of lung, kidney, and thyroid give rise to bone metastases in approximately 30-40% of cases [1]. In contrast, gastrointestinal tract carcinomas rarely produce bone metastases (<10%) [3]. The primary problems that arise from bone metastases are pain, pathological fractures, spinal cord compression, hypercalcemia, and bone marrow suppression [1]. Decalcification ProcessBone, a specialized connective tissue, is composed of a collagenous and non-collagenous matrix proteins strengthened by deposits of calcium salts that play an essential role in the hard, rigid architecture and function of bone [3,4]. In order to study bone pathology, bone tissue is routinely placed within a formalin fixative followed by a decalcification procedure to remove the calcium salts. If a bone specimen is placed into a decalcification solution without prior formalin fixation the tissue will be macerated destroying tissue morphology and nuclear staining [5]. Removal of the calcium salts makes the bone tissue pliable and flexible which is necessary in order to section the tissue for microscopic evaluation. The decalcification procedure can be a lengthy process that may take several days or even weeks depending on the size of the bone. In our pathology department, once the tissue is submitted for decalcification the specimen is recorded into a log book detailing the date and time the specimen is placed into and removed from the Decal solution. Our pathology department uses a Decal solution named Nitrical which contains deionized water and nitric acid. Specimens in Decal solution are checked daily for pliability and softness. The Decal solution is changed on a regular basis due to the fact that the calcium ions that have been removed may saturate the solution preventing further decalcification. Occasionally, extremely rigid tissue may be processed within a microwave histoprocessor in order to decrease the amount of time for decalcification. Experienced histotechnologists test the completeness of decalcification through bending the tissue. Larger, denser bone specimens may be evaluated with X-ray radiographs to determine if the specimen is appropriately decalcified [5]. When the decalcification process is complete, the tissue is thoroughly washed in running tap water for 15-30 minutes to rinse off residual acid and neutralize the tissue. Appropriately size cut sections (0.5 cm in thickness overall size and volume not as critical as thickness) of bone tissue adequately formalin-fixed and decalcified are at this point able to be processed, embedded, and sectioned for light microscopy. All solutions utilized in the decalcification process are changed daily [6]. ResultsFrom January 2011 to August 2011 approximately 3076 specimens were submitted for decalcification according to the log book of which 86 specimens w...

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Influence of decalcifying agents on immunoreactivity of formalin-fixed, paraffin-embedded tissue

Cited by 49 publications

References 16 publications

When Tissue Antigens and Antibodies Get Along

When Tissue Antigens and Antibodies Get Along

Suggested Guidelines for Immunohistochemical Techniques in Veterinary Diagnostic Laboratories

Pathology of Metastatic Tumors to Bone: Effects of Decalcification as Experienced at a Single Cancer Center

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