Automated staining was performed using the DAKO Omnis system according to the manufacturer’s protocol. For antigen retrieval, paraffin-embedded sections were heated in Envision Flex Target Retrieval Solution. Primary antibodies were diluted in Envision Flex Target Retrieval Solution and incubated at 32°C for 1 hour (refer to Key Resources Table). Fluorescent secondary antibodies purchased from Invitrogen were incubated for 30 minutes. Slides were mounted with ProLong® Gold Antifade Mountant containing DAPI as a nuclei counterstain. Images were collected using the Zeiss Axio Observer microscope.
Omnis system
Manufactured by Agilent Technologies
The Omnis system is a modular and automated analytical platform designed for laboratories. It integrates multiple analytical techniques, including spectroscopy and chromatography, to provide comprehensive sample analysis capabilities. The Omnis system is designed to deliver reliable and consistent results, facilitating efficient laboratory workflows.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using omnis system
1
Automated Immunofluorescence Staining Protocol
2
Histologic Diagnosis of Essential Thrombocythemia
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Histologic confirmation of the clinical diagnosis of ET as defined by the 2017 WHO classification was performed by two experienced pathologists (G.A.C and U.G.). Formalin-fixed, paraffin-embedded BM biopsy samples obtained at diagnosis were available for all patients. Sections were stained with hematoxylin-eosin, Giemsa, and Gomori's silver impregnation for evaluation of morphologic features and BM fibrosis. Immunohistochemistry for the evaluation of CD34+ precursors and micro-vessel density was performed via automated Dako OMNIS system, by means of the streptavidin-biotin-peroxidase-conjugated method.
Morphologic parameters were assessed according to the WHO classification, as follows: age-related cellularity, quantitative assessment of granulopoiesis, erythropoiesis and megakaryocytes, left-shifting of granulopoiesis and erythropoiesis (relative increase in immature to intermediate forms or proerythroblasts), presence, size (small vs. large) and density (loose vs. dense) of megakaryocyte clusters, morphologic features of megakaryocytes (hyperlobulated nuclei, cloud-like/bulbous nuclei, naked nuclei, maturation defects), grade of BM fibrosis, presence of osteosclerosis, density of vessels, presence of vascular ectasia and endoluminal hemopoiesis, presence of reactive lymphoid aggregates.
Morphologic parameters were assessed according to the WHO classification, as follows: age-related cellularity, quantitative assessment of granulopoiesis, erythropoiesis and megakaryocytes, left-shifting of granulopoiesis and erythropoiesis (relative increase in immature to intermediate forms or proerythroblasts), presence, size (small vs. large) and density (loose vs. dense) of megakaryocyte clusters, morphologic features of megakaryocytes (hyperlobulated nuclei, cloud-like/bulbous nuclei, naked nuclei, maturation defects), grade of BM fibrosis, presence of osteosclerosis, density of vessels, presence of vascular ectasia and endoluminal hemopoiesis, presence of reactive lymphoid aggregates.
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