Hematology: Manual Differential

What are the components of the Wright's stain (aka Romanowsky stain)?

methylene blue

eosin

glycerin

Methylene blue absorbs onto the (acidic/basic) portion of the cell.

Eosin absorbs onto the (acidic/basic) portion of the cell.

Methylene blue: acidic

Eosin: basic

What granules stain:

orange-pink?

dark blue-black?

dark purple nuclei?

pink-purple granules?

orange-pink: eosinophils

dark blue-black: basophils

dark purple nuclei: segmented WBCs and lymphs

pink-purple granules: neutrophil

What is the purpose of the 6.4 phosphate buffer when staining?

used to determine the pH of the stain

If red cells are too red and the nuclei stain a pale gray-blue, what happened to the Wright stain?

too *acidic*-- excess buffer to stain ratio, insufficient staining time, over-washing

If red cells stain blue and nuclei stain too dark blue, what happened to the Wright stain?

too *alkaline*-- excess stain to buffer, not adequately rinse

What causes a too pink or light Wright stain?

insufficient staining, prolonged washing, too high acidity of the stain or buffer

What cell lines are denoted?

CFU-G

CFU-E

CFU-M

CFU-M

granulocytes

erythroid cells

macrophage/monocyte

megakaryocyte

What are the 6 components of manual differentials?

100 cell differential

RBC morphology

WBC morphology

PLT morphology

WBC estimate

PLT estimate

What objective powers are the following performed on?

perform a 100-cell differential

look for even distribution

perform the WBC estimate

perform the platelet estimate

perform a 100-cell differential= *100x*

look for even distribution= *10x*

perform the WBC estimate= *40x*

perform the platelet estimate= *100x*

What 3 morphologies are included in the differential?

RBC, WBC, PLT

What are the 5 characteristics to be considered for WBC differentiation?

cell size, nucleus, cytoplasm, granules, nuclear:cytoplasm ratio

What are the 3 characteristics to be considered of the nucleus to differentiate the cell?

chromatin/non-chromatin pattern

nucleolus

shape of the nuclear membrane

What 3 considerations need to be made for the cytoplasm characteristics?

color, inclusion, vacuoles

What does PMN stand for?

polymorphonuclear cells (segmented neutrophils)

What is the function of the neutrophil?

phagocytosis

What conditions are associated with neutrophilia?

increased neutrophil ct

leukmoid reactions, bacterial infections, chronic myelogenous leukemia, myelofibrosis, pregnancy

What conditions are associated with neutropenia?

decreased neutrophil ct

viral illness, malaria, thyroid disorders, aplastic anemia, typhoid fever

What is the shape of the nucleus in a band/Stab shape?

horse shoe (not segmented out yet)

What is the reason for the finding of the band neutrophil in the peripheral smear?

they are not fully mature neutrophils yet-- cells are being pushed out in increased numbers as a response to a condition/disease process

What are the two types of lymphocytes and their functions?

T-cell: cellular immunity

B-cell: antibody production

What conditions are associated with lymphocytosis?

increased lymphs

viral infections, pertussis, toxoplasmosis, TB, syphillus, chronic lymphocytic leukemia

What conditions are associated with lymphocytopenia?

decreased lymphs

AIDS, Hodgkin's lymphoma, lupus, renal failure, certain drugs

What cell is generally used to compare the size of a normal lymphocyte?

about the same size as an RBC

What is the main function of the monocyte? During this process, what can be observed in this cell?

phagocytosis; vacuoles can be observed

T/F: eosinophilic granules are secondary granules

true

What is the function of eosinophils?

enhance or suppress the immunoinflammatory reaction, also mildly phagocytic

What conditions are associated with eosinophilia?

increased eosinophils (NAACP)

neoplasms

asthma

allergic

connective tissue disorders

parasites (esp. pinworm)

(CML, polycythemia vera, myeolofibrosis, Hodgkin's lymphoma, essentail thrombocytopenia, allergic disorders, parasitic infections)

What conditions are associated with eosinopenia?

shock, trauma, surgery, corticosteroids, epinephrine

What two chemicals do the granules of the basophil contain?

heparin and histamine

What is the function of the basophil?

enhance immunoinflammatory reaction

What conditions are associated with basophilia?

increased basophils

allergic reaction, chicken pox, ulcerative colitis, various myeloproliferative disorders (CML, polycythemia vera, myelofibrosis)

What is the difference between relative range and absolute range?

relative range is the % specific WBC to total WBC ct.

absolute range expresses the quantification of the specific WBC type

What is the normal range for WBCs?

5,000-10,000 /uL

What are the relative ranges for the following?

neutrophils

lymphocytes

monocytes

eosinophils

basophils

neutrophils: 50-70%

lymphocytes: 20-40%

monocytes: 1-10%

eosinophils: 0-5%

basophils: 0-2%

Secondary granules show up in which stage?

myelocyte stage