骨髓细胞学检查英文ppt课件(第二部分)

Testing of bone marrow cytology(2)主讲人：Testing of bone marrow cytolog.The content,method and clinical significance of the bone marrow cytomorphologic examination.The content,method and cliThe test of bone marrow slides is carried out as follows:1.low power scan(1)make sure if the material-drawing and the film preparation of bone marrow aspiration is satisfied:The specimen should be consisted of 3 parts:the head,the body and the tail.The test of bone marrow slides(2)judge the extent of bone marrow hyperplasia:The extent of bone marrow hyperplasia is reflected by the amount of karyocytes in bone marrow generally.Observe the ratio of karyocyte to mature erythrocyte under low power.(2)judge the extent of bone maThe extent of bone marrow hyperplasia is posited using the 5 grades classification.The extent of bone marrow hype(3)observe megakaryocytes:Four principal aspects about megakaryocytes should be inspected:1)amount2)degree of maturity3)function of generating platelets4)shape of megakaryocytes and platelets(3)observe megakaryocytes:(4)check whether abnormal cells exist:Abnormal histiocyte Metastatic carcinoma cellGaucher cellNiemann-Pick cellOnce abnormal cells were found,they should be inspected under oil immersion.(4)check whether abnormal cell2.oil immersion assayChoose the boundaries of the films coating body and tail.(1)count the karyocytes differentially:Calculate the ratio of the granulocyte to the erythrocyte(G:E).(2)observe the morphology of cells:Including the shape of mature erythrocyte and whether special abnormal cell and parasitic agents abound.2.oil immersion assay3.the clinical significance of the resultThe extent of bone marrow hyperplasia:(1)extremely active hyperplasiaReflect the hyperfunction of bone marrow hematogenesisConditions:leukemia,esp.chronic myelocytic leukemia(CML)3.the clinical significance o(2)the manifestly active hyperplasiaReflect the vitality of bone marrow hematogenesisConditions:hyperplastic anemia,leukemia,proliferating bone marrow diseases,idiopathic thrombocytopenic purpura(ITP),hypersplenia(2)the manifestly active hyper(3)the active hyperplasiaReflect the basically normal condition of bone marrow hematogenesisConditions:normal bone marrow,hyperplastic anemia,chronic aplastic anemia with local compensatory hyperplasia(3)the active hyperplasia(4)the decreased hyperplasiaReflect the decreased function of bone marrow hematogenesisConditions:chronic aplastic anemia,granulocytopenia,agranulemia,myelofibrosis(4)the decreased hyperplasia(5)remarkably decreased hyperplasiaReflect the nonfunction of bone marrow hematogenesisConditions:acute aplastic anemia,bone marrow necrosis(5)remarkably decreased hyperp骨髓细胞学检查英文ppt课件(第二部分)骨髓细胞学检查英文ppt课件(第二部分)The ratio of cells in each series at each developing phase in bone marrow(1)Granulocyte seriesAlmost 50%-60%of karyocytes are granulocytes.With cell maturity,the ratio of each developing phase increased.Myeloblast1%,promyelocyte5%Myelocytes+metamyelocytessegmented granulocytesMost of the eosinophilic granulocytes and the basophilic granulocytes are usually mature in bone marrow.eosinophilic granulocytes 5%basophilic granulocytes 1%The ratio of cells in each ser(2)Erythrocyte seriesNormoblasts20%Pronormoblast1%Basophilic normoblast5%Polychromatic normoblast10%Orthochromatic normoblast10%G:E is defined as the ratio of the percentage of granulocyte series to erythrocyte series.Reference value:(2-4):1(2)Erythrocyte series(3)Lymphocyte seriesThe percentage of cells of lymphocyte series is approximately 20%of karyocytes.Higher in childhood,it even goes up to 40%.Mature lymphocytes:majorityLymphoblasts and prolymphocytes:rare(3)Lymphocyte series(4)Monocyte series4%,almost all of the cells are mature monocytes.(5)Plasmacyte seriesUsu.2%,mainly mature plasmacytes.(4)Monocyte series(6)Megakaryocyte systemIn a film preparation of 1.5cm 3.0cm,the normal value is defined as 7-35 megakaryocytes can be seen.Granular megakaryocytes and thrombocytogenous megakaryocytes:commonly seenMegakaryoblasts:seldom be seenAmong the megakaryocyte system:Promegakaryocyte5%Granular megakaryocytes 10%-27%thrombocytogenous megakaryocytes 44%-60%Naked megakaryocyte 8%-30%(7)Other cellsA few non-hemapoietic cells can be seen,e.g.reticulocytes,endotheliocytes,tissue basophilic cells,etc.(6)Megakaryocyte systemClinical significance of changes in ratio of each seriesClinical significance of changChanges of ratio of granulocytes and erythrocytes(G:E)Changes of ratio of granulocyt骨髓细胞学检查英文ppt课件(第二部分)骨髓细胞学检查英文ppt课件(第二部分).Cytochemical staining of the blood cellThe cytochemical staining is a kind of assay,based on cell morphology,which uses chemical reaction principles,staining bone marrow slides according to a certain procedure,and at last observing the cytochemistry composition and its changes as seen on the microscope.It can be used in the identification of category of blood cells.Cytochemical staining of th1.Peroxidase staining(POX)Result:no blue or black particles in the cytoplasm:(-)Tiny and diffusely distributed granules in the cytoplasm:weakly positiveThick and dense granules:strongly positiveSignificance:identify the category of acute leukemiaAcute myeloblastic leukemia:strongly positiveAcute monocytic leukemia:weakly positive or(-)Acute lymphoblastic leukemia:(-)POX is most valuable in the identification of AML and ALL.1.Peroxidase staining(POX)Res2.Sudan black B staining(SB)A kind of fat-soluble stain,can make the intracytoplasmic lipid turn to brownish-black or deep black particles.Result:almost the same as POX.Granulocyte series:navemature,the positive reaction becomes strongerMonocyte series:weakly(+)Lymphocyte series:(-)Significance:the same as POX.2.Sudan black B staining(SB)A3.Neutrophil alkaline phosphatase staining(NAP)Result:ALP mainly exists in the mature neutrophilic granulocyte(stab or segmented),other cells(-)Positive result:gray to brownish-black particles in the cytoplasmThe intensity of response can be divided into 5 grades:-,1+,2+,3+,4+.The result is presented as the percentage of the positive cells and the score.Reference valueAdult:positive ratio of NAP:10%-40%,the score:40-80 pointsSignificance:acute festered infectious diseases:remarkably,viral infections:normal of slightlyCML:remarkably,usu.score 0,leukemoid reaction:extremelyAML,ALL,acute monocytic leukemia:normal or Aplastic anemia,PNH3.Neutrophil alkaline phospha4.Acid phosphatase staining(ACP)Result:positive reaction:brownish-black particles in the cytoplasm.Significance:assist in the diagnosis of hairy cell leukemia:hairy cell(+)or strongly(+),the activity can not be inhibited by L-tartaric acid.Identify T lymphocyte and B lymphocyte:T cell(+),B cell(-).Gauchers disease(+),Niemann-Pick disease(-)monocyte(-),histiocyte(-),reticular cell(-),megakaryocyte(-).4.Acid phosphatase staining(A5.Naphthol AS-D chloroacetate esterase staining(AS-D NCE)Also called specific esterase(SE),granulocyte esteraseResult:positive reaction:red sediment in the cytoplasm.SE mainly exists in granulocyte series,but microleukoblast(-)or weakly(+),from promyelocyte to mature neutrophilic granulocyte(+),promyelocyte:strongly(+),activity weakened with the maturity of the cells.Eosinophilic granulocyte,lymphocyte,plasmacyte,normoblast:(-)Monocyte(-)or weakly(+)Significance:AML:strongly(+),acute monocytic leukemia and ALL(-)Acute myeloblastic-monocytic leukemia:cells of granulocyte series(+),cells of monocyte series(-)5.Naphthol AS-D chloroacetate6.Alpha-naphthol acctate esterase staining(-NAE)Also called nonspecific esterase(NSE)or monocyte esteraseResult:gray-black or brownish-black sediment in the cytoplasm.NSE mainly exists in monocyte series.Monoblast(-)or weakly(+),promonocyte and monocyte(+),granulocyte series(-)or weakly(+),lymphocyte(-)Significance:identify the acute monocytic leukemia and AML.6.Alpha-naphthol acctate este7.Staining for glycogenAlso called the periodic acid-Schiffs reaction(PAS)Result:positive:the cytoplasm stains red,red substance can be granular,block-like or diffused red.The degree of positive reaction can be divided into 4 grades:strongly(+),(+),weakly(+)and(-).Myeloblast(-),from promyeloblast to segmented neutrophilic granulocyte(+),the positive strengthened with the maturity of cells.Monocyte:weakly(+),lymphocyte(-)or weakly(+)Normoblast and erythrocyte(-)megakaryocyte and platelet(+),with the maturity of cells,positive reaction strengthened7.Staining for glycogenAlso cSignificance:normoblast of erythremic myelosis or erythroleukemia:strongly(+)7.Staining for glycogenGaucher cell:strongly(+),adenocarcinoma cell:strongly(+)Significance:normoblast of er8.Iron stainingAlso called Prussian blue reactionResult:(1)Extracellular ferritin:observe the ferritin inside monocyte-phagocyte series in the bone marrow parvule(outside the normoblast).Positive:light bluish-green and homogeneous shapeless or pearl-like,coarse grain-like,blue or deep blue,or blue-black block-like substance.The positive reaction can be divided into 5 grades:-,1+,2+,3+,4+.8.Iron stainingAlso called Pr(2)Intracellular ferritin:the ferritin in normoblast.Sideroblast:1 to 5 blue and tiny ferritin granules surrounding the nucleus of normal normoblast(mainly orthochromatic normoblast)Crico-sideroblast:one sideroblast with over 10 thick and deep stained ferritin granules,which is surrounding the nucleus for over 2/3 of the nucleonic perimeter.8.Iron staining(2)Intracellular ferritin:theReference value(1)extracellular ferritin:1+2+,mostly 2+(2)intracellular ferritin:20%90%,the average is 65%Significance:(1)iron deficiency anemia:extracellular ferritin(-),percentage of sideroblast,ofter below 15%,even 0%.(2)non-iron deficiency anemia:including thalassemia,sideroblastic anemia,hemolytic anemia,megaloblastic anemia,aplastic anemia,displacement anemia,the extracellular ferritin,often 3+4+(3)sideroblastic anemia:sideroblast,crico-sideroblast constitutes over 15%of the normoblast.8.Iron stainingReference value8.Iron stainin骨髓细胞学检查英文ppt课件(第二部分).Grouping according to cellular immunologyAlso called cell immunity mark(phenotype)assay.It makes use of monoclonal antibody and immunology techniques to test the specific antigen on the surface of cytomembrane and/or in the cytoplasm.Grouping according to cellu1.Assay methods(1)Immunofluorescence method(2)immunoenzymatic staining1)APAAP(alkali phosphatase anti-alkali phosphatase)2)PAP(peroxydase anti-peroxydase)3)ABC(avidin-bioepiderm-multienzyme complex)1.Assay methods(1)Immunofluo2.The application of cell immunophenotyping(1)recognition of cells in different series(2)Identify different lymphocytes2.The application of cell imm(3)Test the T lymphocyte subgroup(4)Identify cells at various stages of differentiation(5)Identify cells in different functional states(6)Isolation and research cells in different series and stages of differentiation(7)Immunophenotypic analysis of various kinds of leukemic cells(8)Test of the leukemic tiny residue(3)Test the T lymphocyte subg3.The immunophenotypic characteristics of acute leukemia cell(1)The immunophenotypic characteristics of cells in ALL and AML3.The immunophenotypic charac(2)The immunophenotypic characteristics of ALL(3)The immunophenotypic characteristics of AML(2)The immunophenotypic chara.Cytogenetic analysis1.Chromosome analysisThe abnormal clone of chromosome is defined as follows:The same chromosome is tested for structural abnormalities in 2 or more cell division phases.The same chromosome is lost in 3 or more cell division phases.Cytogenetic analysisChromos(1)Denomination of chromosomeThere are 46 chromosomes in one human somatic cell,22 pairs(44)of which are euchromosomes,1 pair(XY)of which is sex chromosome.The 46 chromosomes are divided into Group A-G according to the length and the position of centromere.According to the banding characteristics of each chromosome in differential staining technique,chromosomes are divided into regions and bands.Denomination of chromosomeHuman chromosome fluorescence stainingHuman chromosome fluorescence(2)Karyotyping analysis and writingKaryogram is defined as a complete set of chromosomes in metaphase of cell division which are cut out and ranked according to Danvers system after magnification by photograph.The sequence of karyotype writing:chromosome number,accessory chromosome,chromosomal abnormality,separated by comma.(2)Karyotyping analysis and w(3)Chromosomal aberrationIncluding numerical aberration and structural aberration.There are 2 sets of chromosomes in a normal somatic cell,so it is called a diploid(2n).Chromosomal aberrationeuploidaneuploidhaploidtriploidhypodiploidhyperdiploidtetraploid(3)Chromosomal aberrationChro2.Common chromosomal changes in acute leukemia2.Common chromosomal changes 谢谢谢谢谢谢