Lecture 1

hemostasis system (clotting)

increased leukocyte ct (usually in infection)

reduction in leukocytes

-philia increased neutrophils
-penia reduced neutrophils

increase in immature neutrophils

leukemia-like blood cancer (but not leukemia)

granules of neutrophils that look abnormal due to toxicity

lack of blood and blood components
decreased O2 carrying capacity
iron deficiency

increase in RBC, overproduction of RBC

leukocyte increase abnormally and unable to function normally

extreme reduction in granulocytes
* -osis usually means increase but a- negates the increase, therefore it becomes a reduction (?)

peripheral blood film / smear

multicolored RBC in PBS, indication of RBC being released prematurely from bone marrow

more staining than normal; usually indicating increased immature RBCs (reticulocytes)

variation in color intensity of the stain

variation in cell size as measured by RDW

increase in abnormal RBC of any shape that make up 10%+ of the total population

RBC rupture in peripheral blood

increase in blue stain intensity ? (of basophils?)

study of blood cells (and coagulation system)

5L blood

1. formed elements (cells)
2. liquid portion (plasma)

1. granulocytes
2. mononuclear cells
3. platelets
4. RBCs ?

neutrophils, eosinophils, and basophils

phagocytes, kill bacteria

kills parasites

allergic reactions

release of histamine by basophils

lymphocytes and monocytes

lymphocytes are important to the immune system;
kills tumor cells and make antibodies

phagocytes, kill intracellular pathogens

stop bleeding and help repair vascular injuries

contain hemoglobin which is crucial for O2 delivery form lungs→tissues and transporting CO2 from tissues→lungs

Athanasius Kircher described worms in the blood (parasite or extended WBC)

Antony van Leeuwenhoek gave an account of RBCs
(unsure of what they do, only ID)

Giulio Bizzozero described platelets as petite plaques

James Homer Write developed the Wright stain and opened a new world of visual examination through a microscope

non-nucleated biconcave discs

hemoglobin that contains iron (that binds to O2)

salmon-pink, measures 6-8 microns in diameter with a zone of central pallor

less hemoglobin

count, shape, color, central pallor, size, content
(CSCCSC)

automated or manual using hemocytometer

inclusions (through examination of a stained PBS)

RBC indices from analyzer or PBS examination

PBS examination

RBC indices, PBS examination (Hb content)

anemia and polycythemia

whole blood mixed with Drabkin reagent

solution of potassium cyanide + potassium ferricyanide

Hb gets converted to cyanmethemoglobin

540 nm

ratio of packed RBC volume to whole blood volume (% or L/L)

induces (MCV, MCH, MCHC)

status of bone marrow response to anemia
high: responding efficiently
low: failed, cannot respond to demand

components and blood smear

cell counts, indices, WBC diff, RBC parameters

preparation, estimates, and cell morph

ratio of packed volume of RBCs to the volume of whole blood

45-50%

whole blood taken from veins

smears are made at bedside before blood clots

patient identification

anticoagulant (EDTA lavender tube top)

must be mixed immediately and gently to prevent clotting

within 4 hours of collection

within 5 hours of collection,
kept at room temp until analysis (facility dependent)

too little blood, too much EDTA

1. low blood cts and HCT
2. staining alteration
3. morph changes to RBCs and WBCs on blood film

1. RBC / WBC shrinkage
2. Crenated RBCs
3. WBC membrane damage

+ / - 10% of the stated draw volume

when filling EDTA tube from syringe

prevents proper mixing of sample → insufficient EDTA for blood volume

may lead to platelet clumping and clotting
(clotting sample = rejected!)

cells in EDTA for >5 hrs at room temp

1. artifacts
2. MCV increased
3. HCT increased
4. necrobiotic / necrotic WBCs (decrease WBC cts)
5. progressively vacuolated WBCs (esp. Neutrophils)

crenated RBCs, spherocytes

MCV increased due to RBC swelling

increased HCT due to increase in MCV

storage at 4C for up to 12 hours to minimize effects of overexposure to EDTA

slow / difficult draws or unproperly mixed tubes (blood many not get exposed to EDTA in time)

small clots or clumping platelets may affect pt results

must make PBF

may be unexpectedly (falsely) decreased

observing for obvious clots in tube

polychromatic stain

eosin Y and methylene blue

basic stain that stains acidic cellular components from blue-grey to dark blue

RNA. DNA, basophil granules

acidic stain that stains basic cellular components from light to deep pink to red or orange in color

hemoglobin and eosinophilic granules

neutrophils have cytoplasmic granules with neutral pH and pick up both stains (thiazine-eosinate complex)

appear pink / violet

1. stain / smear quality
2. sample integrity
3. WBC and PLT estimates
4. WBC differential
5. abnormal WBCs or reactive lymphs
6. alterations in RBC and PLT morph
7. look for parasites, bacteria

check for PLT clumps, fibrin strands, cell distribution, etc

to find correlation with analyzer report in case of flags

RBC diameter (volume) on a Wright stained smear

average size of RBC

80-100 fL (10^-15 L)

between microcytic (MCV <80), normocytic (MCV 80-100) and macrocytic (MCV >100) anemia

weight of the hemoglobin in the average RBC

26-34 pg (10^-12 g)

concentration of Hgb in average RBC

32-36 g/dL (Hgb/Hct)

hypochromic (MCHC <32) and normochromic (MCHC 32-36) anemias

degree of volume variation

variation in diameter (anisocytosis)

variation in size of RBC as reflected by RDW

the RDW is low

high cell size variation

between anemias with minimal anisocytosis

12-14.5

young RBCs that contain RNA

1.5-2.5%

slightly larger in diameter than RBC and stain slightly blue-gray with Wright stained smear
(polychromasia)

RNA is broken down and erythrocytes become normochromic

suggest bone marrow is responding appropriately to anemia

supra vital staining (methylene blue) reticular pattern on PBS

relative count, absolute ct, immature reticulocyte fraction (IRF) using fluorescence-based tech

total number of leukocytes in blood

adult: 4.5-11 x10/L
child over 1: 5-17
newborn: 9-30

decreased WBC count

increased WBC count

segmented neutrophils
band neutrophils
eosinophils
basophils
lymphocytes
monocytes

infection and inflammation
leukemia

neutrophils 62%
lymphocytes 30%
monocytes 5%
eosinophils 2.5%
basophils 0.75%

150-450 x10^3L

splenomegaly, disseminated intravascular coagulation, bone marrow failure

reactive thrombocytosis (iron-deficiency anemia) and essential thrombocythemia

stop blood system (coagulation)

coagulation factors
platelets
vasculature
fibrinolytic system

aspirates
biopsies

myeloperoxidase
Sudan black B stain
terminal deoxynucleotidyl transferase
leukocyte acid (TRAP) and alkaline phosphatase

myeloperoxidase stain
Sudan black B stain
TRAP and Alkaline phosphatase

terminal deoxynucleotidyl transferase

molecular genetic tests

karyotyping (Philadelphia chromosome translocation t(9;22)

CSF, synovial, pericardial, plural, and peritoneal

always STAT, always acute