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What cell types does Myeloperoxidase stain?
Primary granules containing peroxidase; Promyelocytes, myelocytes, metamyelocytes, PMN/Band
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What cell types does Sudan Black stain?
Primary and Secondary granuls, all granulocyte precursors. Differentation of ALL from AML
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What cell types does napthol AS-D Chloroaetate Esterase stain?
Myeloblast primary granules; to a lesser degree PMN, Basos, and Mast cells; Eos, Monos, and Lymph stain negative.
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Alpha-Naphthyl butyrate or Alpha Naphthyl acetate esterase stain which types of cells?
Monoblasts; monocytes, hisitocytes, macrophages, and megakaryoblasts. Granulocytes stain negative.
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TdT or Terminal deoxynucleotidyl transferase stains what cell type?
- Stem Cells, Immature lymphs in BM, DX Lymphoblastic leukemias
- Not found in Mature B cells
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Acid Phosphatase stains what cell types?
T-Lymphoblasts
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PAS (Periodic Acid-Schiff stains what types of cells?
Ereythroblasts; glycogen and glycoproteins; DX erthroleukemia
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What stains Ereythroblasts?
PAS, Periodic Acid-Schiff
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What stains B- Lymphoblasts?
TRAP- Acid Phosphatase; good for hairy cell leukemia
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What stains B-lymphoblasts?
TRAP-Tartrate resistant acid phosphatase
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What stains T-Lymphoblasts?
Acid Phosphatase
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What stains immature lymphs?
TdT- Terminal deoxynucleotidyl transferase - DX lymphoblastic leukemias
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What stains monoblasts?
Alpha Napthyl butyrate or Alpha naphthyl acetate esterase, negative for ganulocytes, positive for monoblasts/monocytes/M0/megakaryoblasts
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What stain is used for myeloblast primary granules?
Napthol AS-D Chloroacetate esterase; stainsmyeloblasts, PMN, Basos, and mast cell primary granules; not Eos, Monos, or lymphs
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What is used to stain all granulocyte precursors, both primary and secondary granules?
Sudan Black; differentiates AML from ALL
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What stain is used on primary granules that contain peroxidase?
Myeloperoxidase; MPO, stains promeyelocytes, myelocytes, metamyelocytes and PMN/Band
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Hunter's and Hurler's (MPS)
Mono azurephilic granules, accumulations of vacuoles containing MPS mucopolysaccharides
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Tay-Sachs
Vacuolated lymphocytes; foam cells
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Neimann-Pick
Histiocytes packed with lipids; foam cells and "sea blue histiocytes"
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Gaucher's
Monocyte/M0/Histiocyte have crumpled tissue paper cytoplasm
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May-Hegglin anomaly
Dohle bodies, giant platelets, thrombocytopenia
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Chediak-Higashi syndrome
- Hepatospleenomegaly
- lymphs and monos have a single red granule
- Giant gray/green cytoplasmic granules in PMN
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Alder's anomaly - Alder-Reilly
- Coarse dark granules in WBCs (all lines possible)
- Lipid deposits in cytoplasm
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Pelger-Huet anomaly
- Heterozygoes - dumb-bell nucleus
- homozygotes - round nucleus
- "pince nez" cells
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M0/Monocyte/Histiocyte have crumpled tissue paper cytoplasm
Gaucher's
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Histiocytes packed with lipids; foam cells present, and can see "sea blue histiocytes"
Neimann-Pick
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Vacuolated lymphocytes; foam cells
Tay-Sachs
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Monos have azurephilic granules and there are MPS accumulations
Hunter's and Hurler's (MPS)
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Pince nez cells
Pelger-Huet
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Coarse dark granules in all WBCs
Lipid deposits in cytoplasm
Alder's -Reilly
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Giant gray/green cytoplasmic granules
Lymph and mono have single red granule
hepatosplenomegaly
Chediak-Higashi syndrome
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Thrombocytopenia
giant platelets
dohle bodies in PMN
May-Hegglin anomaly
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If: WBCs too high
Then dilute, if 3x rule is on, WBCs probably ok
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If: Instrument ++++WBCs
Dilute, rerun, multiply
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If: Ictirus/Lipemia
- Hgb, MCH, MCHC increased and RBC low
- Then: look at slides, replace plasma with saline and/or notify physisian
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If: MCHC, MCH, and Hgb high, and RBC low
Then: Ictirus or Lipemia
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If: Cold Agglutenin
- Then: Low RBC, High MCV, MCHC
- Warm to 37 and rerun
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If Low RBC, High MCV and MCHC
Then: Cold agglutenin
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If: increased WBC count, and low platelet count
Then: could be satellite platelets; look at slide, redraw in blue top and multiply result by 1.1 for the 10% dilution
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Spleen Functions
- 1. Hematopoiesis 2-7mo fetal
- 2. Granulocyte and platelet storage
- 3. Culling fo old WBCs
- 4. Processing of Blood Borne pathogens
- 5. Part of the RES and a lymph organ
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MCV
- Mean Corpuscular Volume
- 80-100fL, HCT/RBC x10
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MCH
- Mean Corpuscular Hgb
- 27-31pg
- Hgb/RBC x10
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MCHC
- Mean Corpuscular Hgb Concentration
- 32-36%
- Hgb/HTC x100
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RDW
- RedCell Distribution Width
- 11.5-14.5%
- Anisocytosis
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Hgb
- 14-18 Males
- 12-16 Females
- g/dL
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HCT
- 42-52% Males
- 37-47% Females
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WBC Normal Numbers
4-11 x10^9g/L
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WBC Diff
- Segs -50-70
- Band- 2-6
- Lymph - 20-44
- Mono 2-9
- Eos 0-4
- Besos 0-2
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RBC
- 4.7-6.1 Males
- 4.2-5.4 Females
- x10^12/L
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Anemic Color Classification
- MCHC
- Normochromic- 32-36%
- Hypochromic- <32%
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Anemic Size Classification
- MCV
- Microcytic- <80fL
- Normocytic- 80-100fL
- Macrocytic- >100fL
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WBC Diff Count
- 10x Low
- Average 5 fields x1000 /4
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LAP (Leukocyte alkaline phosphatase) Stain
- Chronic Myeloproliferative Disorders (CMPDs)
- ---not high in CML
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Stain for CMPDs
LAP (Leukocyte alkaline phosphatase) Stain
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CML
- Phidelphia chromosome; 9:22 translocation
- Clonal myeloproliferative disorders - Granulocytes
- Myeloid Leukocytosis- all stages of development
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3 Phases of CML
- Chronic- huge increased WBC count
- Accelerated- 6-18mo long
- Blast Crisis- looks like myelogenous or acute lymphoblastic leukemia
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Polycythemia Vera
- Old people
- Hypercellular BM
- Overproduction of mature RBCs and Platelets
- JAK 2 mutation
- More sensitive to EPO
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PV RBC description
- Microcytic hypochromic, exhausting of Fe
- Blood EPO levels are low
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PV Diagnosis
- JAK2 mutation, or other
- Thromobocytosis > 400,000/uL, WBC>12,000/uL
- Low serum EPO
- Treatment: Phlebotomy
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AML causes
Chromosomal translocation
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M1
Myeloblastic -without differentation
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M0
Myeloblastic - minimally differentiated
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M2
Myeloblastic with maturation
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M3
Promyelocytic leukemia
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WHO AML Classification
- I - Cytogenic abnormalities
- II - Multilineage dysplasia
- III - Myelodysplastic syndromes (therapy)
- IV - Not categorized
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ALL
- Acute lymphoblastic leukemia- CD10 abd TdT positive, >100 x 10^9/L
- Anemia
- Thrombocytopenia
- Neutropenia
- Overpopulation of lymphoblasts - often clefting
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Mononucleosis, Cytomegalovirus, Acute Viral Hepatitis
Increased WBC, Decreased PMN, Increased lymphs, Reactive lymphs on smear
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Mononucleosis
- Reactive T-cells react to infected B-cells
- EBV infected cells can have thrombocytopenia or anemia due to Ab (IgG IgM)
- >10% reactive lymphs
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CMV
- Similar to EBV
- IgG and IgM Ab to CMV Ag
- Reactive lymphocytes with negative heterophil antibody
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Acute Viral Hepatitis
- Hepatitis Ab findings diagnostic
- Absolute lymphocytosis and reactive lymphs
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