Hematology 1 - Cytochemical Stains

HEMATOLOGY 1 – CYTOCHEMICAL STAINS STAIN Leukocyte Alkaline Phosphatase (LAP) CLASSIFICATION Enzymatic Myeloperoxidase Enzymatic Periodic Acid Schiff (PAS) Nonenzymatic Terminal Deoxyribonucleotidyl Transferase Nonenzymatic PURPOSE Used to detect alkaline phosphatase activity in the neutrophils and bands; used to differentiate a leukemoid reaction from chronic myelogenous leukemia Useful in differentiating acute myeloid leukemia and acute lymphoid leukemia. Also useful in diagnosing congenital deficiency of neutrophil myeloperoxidase. Recognizes some cases of erythroleukemia and acute lymphoblastic leukemia. STRUCTURES STAINED Cytoplasm of bans and segmented neutrophilic granulocytes. Glycogen or glyco-containing cellular elements. POSITIVE for AML M6 erythroleukemia, Gaucher Disease, Neoplastic lymphoblasts, neoplastic myeloblasts. Used to identify primitive myeloid leukemia cells, TdT is considered a marker for primitive lymphoid cells (Lymphoblasts) DNA polymerase, immunoperoxidase POSITIVE for myeloid leukemia (AML) Myeloperoxidase is present in primary granules of myeloid cells. Early meyloblasts are negative, with granular positivity appearing progressively as they mature. INTERPRETATION Leukocytes are scored (0 to 4+) corresponding with the intensity of blue color; the sum of scores is called LAP score; which results from 0 to 400, with a normal score being between 20 and 100. A score higher than normal may indicate leukemoid rection. POSITIVE for neutrophilic granulocytes except blast forms. NEGATIVE for basophils, lymphocytes, and erythrocytes. NEGATIVE for leukemia (ALL) acute lymphocytic REMARKS Also known as Neutrophil alkaline phosphatase (NAP). Use of EDTA is NOT recommended because it inhibits enzyme activity. Azide is used as an inhibitor of MPO. It is most abundantly seen in neutrophil granulocytes and antimicrobial activity. Reacts with aldehyde groups in glycogen, mucoproteins, and other high molecular weight carbohydrates. Glycogen is predomiant in leukocytes. Catalyzes the irreversible addition of deoxynucleotides to the 3’-hydroxy groups on the end of DNA; marker for immature leukocytes; Estrase Enzymatic Used in hematopathology to differentiate the monocytic lineage from lymphoid and other myeloid lineage cell types. Monocytes and their precursors stain a red color with this stain. Sudan Black B Nonenzymatic Useful in differentiate AML and ALL. Stains certain phospholipids and lipoproteins. It stains positive in granulocyte and monocytes-macrophages and negative in lymphocytes Perl’s Prussian Blue Nonenzymatic Iron Tartrate-Resistant Acid Phosphatase Enzymatic Determines the amount and location of iron, an important red cell component, stores in the body that aids in diagnosis of hematologic disorders. Detectes tartrate-resistant acid phosphatase in hairy cell leukemia. Granules in lymphocytes. Transes Material S.Y. 2023-2024 – CREATED BY IAMNUMBEREIGHT | The struggle is real and so is God. Non-specific esterase-staining mononuclear phagocytes are easily distinguished because of red or reddish brown staining pattern produced in esterase-positive cells. The cells that do not have non-specific esterase activity appear unstained. Myeloblasts are negative. Neutrophil precursors contain increasingly numerous granules. Monocytes contain slight to moderate scattered positivity that may be obscured by the counterstain. It is useful in Acute Leukemia, M1, M2, M3, M4. M5a, and M5b POSITIVE for thalassemia, alcoholism liver disease, lead poisoning, anemia. In hairy cell leukemia, majority of leukemic cells show positivity in the presence of tartaric acid. differentiates AML from ALL Use of substrate component for stain identity is thought to be a more precise manner to communicate which stains were performed. User must be aware that there may be false-positive reactions in disorders characterized by cytoplasmic lipid vacuoles, such as Burkitt’s lymphoma. Its reaction precipitates free iron into small blue or blue-green granules in RBCs. Free iron is NOT identifiable on Wright or WrightGiemsa-stained blood smears. None. HEMATOLOGY 1 – CYTOCHEMICAL STAINS Toulidine Blue Nonenzymatic Useful for enumeration of basophils and mast cells. Acid mucopolysaccharides in granules of basophils and mast cells. Transes Material S.Y. 2023-2024 – CREATED BY IAMNUMBEREIGHT | The struggle is real and so is God. Granules of basophils and mast cells stain bright red and purple. Nuclei stain blue and cells abundant in RNA may show a blue tint to the cytoplasm. Malignant basophils or mast cells may not show positive reaction.