Manual Differential (Exam 2)

components of Wright's stain

methylene blue

eosin

glycerin

Methylene blue

absorbs onto the acidic portion of the cell

a staining dye/indicator that interacts with nucleic acid molecules and proteins, turning them to a very dark blue color

Eosin

Absorbs onto the basic portion of the cell

eosinophils

granules that stain orange-pink in a good stain

basophils

granules that stain dark blue-black in a good stain

segmented WBCs and lymphs

nuclei that stain dark purple in a good stain

neutrophil

granules stain pink-purple in a good stain

Determine pH of the stain

Phosphate buffer purpose in staining

characteristics of cell staining when stain too acidic

RBCs will be too red

Nuclei will stain pale gray-blue

Caused by excess amount of buffer to stain, insufficient staining time, over-washing

characteristics of cell staining when stain too basic

RBCs stain blue

causes: excess stain to buffer, not adequately rinsed

causes of too pink/light stain

Insufficient staining

Prolonged washing

Too high acidity of the stain/buffer

six components of manual differentials

100-cell differential

RBC morphology

WBC morphology

Platelet morphology

WBC estimate

Platelet estimate

10x

Objective magnification that is used to look for even distribution in the differential procedure

40x

Objective magnification used to perform the WBC estimate in the differential procedure

100x

Objective magnification used to perform a 100-cell differential and perform the platelet estimate in the differential procedure

RBCs, WBCs, Platelets

three morphologies included in the differential

five characteristics to be considered for WBC differentiation

Cell size

Nucleus

Cytoplasm

Granules

Nuclear: Cytoplasm ratio

Chromatin/non-chromatin pattern, nucleolus, shape of nuclear membrane

three characteristics to be considered of the nucleus to differentiate the cell

Color, inclusions, vacuoles

three considerations need to be made for the cytoplasm characteristics

Nuclear to cytoplasmic ratio

Amount of space occupied by the nucleus in relationship to the space occupied by the cytoplasm

Differs from cell size

Polymorphonuclear

PMN

pertaining to a white blood cell with a multi-lobed; neutrophil

phagocytosis

Function of neutrophil

A type of endocytosis in which a cell engulfs large particles or whole cells

Neutrophilia

increased neutrophil count

Conditions associated: Leukemia reactions, bacterial. infections, chronic. myelogenous leukemia, myelofibrosis, pregnancy

Neutropenia

decreased neutrophil count

Conditions associated: viral illness, malaria, thyroid disorders, aplastic anemia, typhoid fever

Horeshoe

shape of the nucleus in the band neutrophil

Band neutrophil

immature neutrophil

seen in peripheral smear when cells are being pushed out in increased numbers as a response to specific conditions

T-cell lymphocytes

cellular immunity

B-cell lymphocytes

antibody production

Lymphocytosis

increased lymphocyte count

Conditions associated: viral infections, pertussis, toxoplasmosis, TB, syphilis, chronic lymphocytic leukemia

Lymphocytopenia

decreased lymphocyte count

Conditions associated: AIDS, Hodgkin's lymphoma, lupus, renal failure, administration of certain drugs

RBC

cell generally used to compare size of normal lymphocyte

monocyte

an agranulocytic leukocyte that performs phagocytosis to fight infection (Vacuoles can be observed during phagocytosis)

cytoplasm: blue/purple

Nucleus: purple

Monocytosis

increased monocyte count

Conditions associated: recovery phase of acute infection, ulcerative colitis, SLE, RA, RB, syphilis, and many hematologic neoplasms such as myelodysplastic syndrome

True

True/False

Eosinophilic granules are secondary granules

Eosinophils

a white blood cell containing granules that are readily stained by eosin

Function: enhance/suppress the immunoinflammatory reaction, mildly phagocytic

Eosinophilia

increased eosinophil count

Conditions associated: CML, polycythemia vera, myelofibrosis, Hodgkin's lymphoma, essential thrombocytopenia, allergic disorders, parasitic infections

Eosinopenia

decreased eosinophil count

Conditions associated: shock, trauma, surgery, administration of corticosteroids/epinephrine

purple-black

color of the secondary granules in the basophil

heparin and histamine

two chemicals contained in the granules of the basophil

Basophil

white blood cell containing granules that stain blue

Function: enhance the immunoinflammatory response

Basophilia

increase in basophils

Conditions associated: allergic reactions, chicken pox, ulcerative colitis, various myeloproliferative disorders (CML, Polycythemia vera, myelofibrosis)

50-70%

relative range of neutrophils

20-40%

relative range of lymphocytes

1-10%

relative range of monocytes

0-5%

relative range of eosinophils

0-2%

relative range of basophils

5000-10000/uL

normal range of WBC

myelocyte stage

Stage of granulopoiesis where secondary granules appear