Advanced Diagnostic Techniques

Ramos‐Vara, José A.; Avery, P.; Avery, Anne C.

doi:10.1016/b978-1-4557-4083-3.00017-6

Cited by 13 publications

(11 citation statements)

References 151 publications

(134 reference statements)

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“…In following these rules, the current optimization process followed the quality assurance ICC guidelines approved by the ASVCP and those for immunohistochemistry . Validation of the antibodies used in this study for canine and feline specimens was determined with previous literature reports or through author experience with immunohistochemistry of formalin‐fixed tissues and ICC of unstained cytologic samples …”

Section: Discussionmentioning

confidence: 99%

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Optimized immunocytochemistry using leukocyte and tissue markers on Romanowsky‐stained slides from dogs and cats

Raskin

Vickers²,

Ward³

et al. 2019

Veterinary Clinical Pathol

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Background: Romanowsky staining is often the initial method used to stain hematologic and cytologic materials. While immunocytochemistry (ICC) is a well-established method on air-dried smears, there are rare veterinary reports of ICC involving Romanowsky-stained slides.Objectives: This study aimed to compare immunoreactivity of unstained vs Romanowsky-stained specimens, evaluate reactions over time, and assess ICC associations with confirmatory tests of 50 lymphoma cases. Another goal aimed to optimize manual ICC protocols with cellular and tissue immunomarkers to detect CD3ε, CD20, Pax5, MHCII, lysozyme, MUM1, vimentin, cytokeratin, and Melan-A antigens on Romanowsky-stained specimens. Materials and methods:Cytologic specimens from cases of lymphoid and nonlymphoid neoplasms were stained with a methanolic Romanowsky method. Additional unstained slides from these cases were used for comparison with the stained materials. Antigen retrieval involved a citrate buffer pH6 or Tris/EDTA pH9 at 95°C for 25 minutes in a decloaking chamber. Immunocytochemistry used known positive and secondary antibody-only negative cytologic controls. Immunoreactivity of unstained and prestained lymphoma slides was graded by the intensity and percent of stained cells. Signal grading was monitored over time for diagnostic differences.Results: Unstained and Romanowsky-stained slides had similar membrane/cytoplasm graded reactions, but unstained slides produced stronger signals. Romanowskystained blood films from B-cell and T-cell leukemias showed minimal loss of signal when monitored over 20 weeks. Signal differences did not change the diagnosis. There was a significant association between ICC and confirmatory tests. Optimization involved antibody dilution and antigen retrieval methodology for each antibody tested. Conclusions:Immunocytochemistry of Romanowsky-stained material can be successfully performed using antibodies against CD3ε, CD20, cytokeratin, lysozyme, Melan-A, MHCII, MUM1, Pax5, and vimentin. K E Y W O R D Santigen retrieval, CD20, CD3, lymphoma, lysozyme, Melan-A, MHCII, Pax5 | 89 RASKIN et Al.

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Section: Discussionmentioning

confidence: 99%

“…Immunocytochemistry (ICC) is routinely performed on blood and cytologic specimens for phenotyping lymphoid neoplasms in veterinary medicine and works well on unstained slides . Round cell neoplasms might be difficult to distinguish by morphology alone and would benefit from advanced diagnostic testing .…”

Section: Introductionmentioning

confidence: 99%

Optimized immunocytochemistry using leukocyte and tissue markers on Romanowsky‐stained slides from dogs and cats

Raskin

Vickers²,

Ward³

et al. 2019

Veterinary Clinical Pathol

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“…Storage times before antigen loss is limited, 32 maintaining a consistent bank of positive control slides is difficult, cell numbers can be low, and the amount of background stain can be high. 33 In addition, as the number of slides could be limited, further material is unlikely to be available from the sample for future diagnostic investigations, for example, further immunostaining. 31 The limitations of ICC have led to the introduction of cell block techniques that allow the advantages of IHC to be combined with a relatively noninvasive method of collection.…”

Section: Discussionmentioning

confidence: 99%

Comparison of effusion cell block and biopsy immunohistochemistry in mesothelial hyperplasia, mesothelioma, and carcinoma in dogs

Milne

Piviani

Hodgkiss-Geere

et al. 2021

Veterinary Clinical Pathol

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Background: Determining the cause of effusions is challenging and might require a biopsy. Whether cell blocks from effusions are representative of biopsies requires investigation. A previously developed immunohistochemical panel aids in the differentiation of hyperplastic and neoplastic mesothelium in canine biopsies but has not been investigated in effusions. Objectives:The study aimed to assess cell blocks as an alternative to biopsies and determine whether immunohistochemistry helps distinguish hyperplastic mesothelium, mesothelioma, and carcinoma.Methods: Effusions and biopsies were collected from five dogs with mesothelial hyperplasia (group MH), six with mesothelioma (group M), and five with carcinoma (group C). Immunohistochemistry (IHC) for cytokeratin, vimentin, Wilm's tumor protein 1 (WT1), desmin, glucose transporter 1 (GLUT1), and insulin-like growth factor II mRNA-binding protein 3 (IMP3) was performed. Sections were scored for staining intensity and the percentage of positively stained cells. Results:In paired cell blocks and biopsies, vimentin and WT1 staining were positively correlated for intensity and the percentage of positive cells, although not all paired results were identical. The intensity of IMP3 staining in cell blocks was higher in group M than in group C (P = 0.012), and WT1 staining was higher in group MH than in group C (P = 0.020). For biopsies, the intensity of WT1 staining was higher in group MH than in group C (P = 0.031). In group C, WT1 was negative in all cell blocks and biopsies, and desmin was negative in four of five cases. Conclusions:IHC results for the cell blocks and biopsies were comparable for potentially useful markers, such as WT1, which helped discriminate between groups. IHC provided additional information, although results were not always definitive. Further studies on a larger population are required.

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“…A malignancy of lymphoid or myeloid origin was also considered but judged less probable. Immunohistochemistry for mammalian hematopoietic cells, such as CD3/CD79a immunostaining for lymphocyte markers, and CD34 immunostaining for hematopoietic cell origin, could be helpful in supporting a cell line of origin; however, immunohistochemistry is not yet validated in amphibians . The sex of the frog was unknown at the time of cytologic evaluation, and therefore, a dysgerminoma or a seminoma were both considerations.…”

Section: Discussionmentioning

confidence: 99%