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445 Cards in this Set
- Front
- Back
Hemato / poiesis
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blood production
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Blood cell production normally occurs in the__________ and is carefully regulated
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bone marrow
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The ability of the cell to go through cell cycle and make more cells
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cell proliferation
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The ability to become a functional cell
Dedicated to a cell lineage RBC, WBC, PLT |
cell differentiation
|
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Orderly process, identified by cell morphology
Carefully regulated |
functional maturation
|
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Normally occurs in fetal development or
During disease states in adult life when bone marrow cannot provide enough red cells |
extramedullary hematopoesis
|
|
splenic
hepatic both are what? |
spleen
liver extramedullary hematopoesis |
|
medullary hematopoesis occurs where?
during what ages of child? during what ages and where on a adult? |
Bone marrow
Primary site at birth 100% BM involvement until 4 yrs when fatty tissue begins to replace active BM Limited to specific bones in adults pelvis, vertebrae, ends of ribs, femur, humerus, sternum, skull |
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adult marrow is what percent adipose and what percent cellular?
what about older adult? |
50:50
older adult 70adipose and 30 cellular |
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descibe living tissue
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- soft, spongy
- very vascular - rich blood supply |
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what is a precursor to platelets
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megakaryocyte
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what makes up 5% of all cells in the blood
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hematopoietic stem cell
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what part of the BM controls the growth
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extracellular matrix
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________ _____ is released from functional cells and causes further stem cell proliferation
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c-kit ligand
|
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chemical signals
May be lineage specific Are synergistic |
cytokines
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binding sites for chemical signals
Are found on the cell surface Bind cytokines and growth factors |
Receptors
|
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Hormones
c-kit ligand is released from functional cells and causes further stem cell proliferation May be specific (EPO, G-CSF) |
Growth factors
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stem cells have what antigen
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CD 34
|
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Morphologically resemble a lymphocyte
high N:C light blue cytoplasm purple, open nucleus |
stem cell
|
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mitotic cycle
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Overall cell size doubles
State of proliferation determines relative cell size Resting state has condensed, non-active DNA resting, g1, s, g2, m, resting |
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how much blood do we have
|
6-8L
|
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plasma is composed of what and is what %
|
water, ions, nutrients
(55-60% of total blood volume) |
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plasma minus clotting factors
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serum
|
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Red cell mass measured as HCT
Spun HCT= what? |
Spun HCT= (100 – [plasma + Buffy coat])
|
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RBC
size and life span |
6-8 microns in size
Life span = 120 days |
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what does hemoglobuin do?
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binds to oxygen
carries H+ ions (buffers blood) carries some carbon dioxide |
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leukocyte size
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8-25 microns
|
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thrombocyte
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platelet
|
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what are platelets- what do they do?
whats there size and lifespan |
cytoplasmic fragments that are produced in BM
Function in hemostasis and survey the vascular system to keep it intact Provide a reaction surface for attachment of circulating clotting factors Form a stable fibrin/platelet plug Size 2-3 microns Life span = 7-10 days |
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what hematology test can be preformed from a EDTA or WB and with a cappilary specimen?
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CBC
Hgb & Hct Indices, RBCM Differential Reticulocyte count Platelet count |
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what hematology tests can be preformed with citrated plasma and not a capp specimen?
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Protime, aPTT
Factor Assays |
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What hematology test can be preformed with an EDTA, WB, or citrated plasma but can not be a capp specimen?
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ESR
|
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what kinda of stain is used for a PBS
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wright
|
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what kind of specimen can you use with PBS and where should they be collected?
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use a EDTA, whole blood specimen
venous capillary |
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what two tests should you not use with a PBS and why?
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Heparin
distorts morphology clumps platelets Na citrate causes a dilutional error |
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Blood smears should be made within __ hours
Smears made after ___ hours are unacceptable |
Blood smears should be made within 2 hours
Smears made after 4 hours are unacceptable |
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Good for bone marrow comparisons and multiple stains
|
coverslip smears
|
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what angle should you have with wedge smear
|
35-45
|
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what is WRight stain composed of
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Polychrome methylene blue
Eosin Methanol (absolute) Potassium and sodium phosphate buffers Na2HPO4 buffer KH2PO4 buffer pH = 6.4-6.8 |
|
Segs & bands
(neutrophils) Lymphocytes Monocytes Eosinophils Basophils |
Segs & bands ~60%
(neutrophils) Lymphocytes ~35% Monocytes ~10% Eosinophils ~ 3% Basophils ~0-1% |
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with a too acidic stain what do the following look like?
nucleus cytoplasm rbc eosinophils |
Nuclear characteristics are pale blue
Cytoplasm looks gray instead of blue Red cells appear bright red/orange Eosinophils granules are brilliant red-orange Cells lack contrast |
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what would cause the stain to be to acidic
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Stain time too short
Extended buffer time Excess washing Old stain Buffer or wash water have acidic pH |
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what would an alkaline stain look like?
nucleus? neutrophil? rbc? eosinophils? lymphocyte? |
Nuclear details appear deep, dark, purple-blue with little contrast in chromatin pattern
Neutrophil granules appear large and toxic Red cells are blue-green Eosinophil granules are blue-gray Lymphocyte cytoplasm is gray-lavender |
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what would cause an alkaline stain?
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Smear too thick
Extended staining time Insufficient washing Buffer or water too alkaline Fresh stain, not aged |
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causes of stain precipitate?
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unfiltered stain
PBS dried flat excessive drying during staining process insufficient washing dirty slide |
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for a PBS at 10x what should be done?
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Choose the correct area
50% of red cells should be touching RBC morphology Rouleaux Agglutination Cell size, shape, color Check feathered edge for platelet clumping Look for parasites-microfilaria at edges COUNT WBC |
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1 WBC (LPF) = ? cells/uL
|
200
|
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what should be done when using 100X
|
Immersion oil has the same refractive
index as glass so no light is lost by refractive scatter Higher power objectives require more light Condenser up & fully open, field aperture fully open |
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what is done on HIGHPOWER
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WBC DIFF
RBC morph platelet estimate |
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1 platelet x 15 K = units?
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1 platelet x 15 K = plt/uL
|
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to get the absolute cell count what do you do?
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Absolute Cell Count = (Relative %) x WBC count/uL
wbc you got from 10x relative if from WBC diff |
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Coarse, dense chromatin
Fine pink granules Life span = hours pink to tan color |
neutrophils
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when are banded neutrophils seen
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under stress
|
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neutrophil ranges:
relative absolute |
Relative 47-80%
Absolute 1800-7700 cells/uL |
|
cytoplasm scant
clear to medium blue cytoplasm dark, dense, lumpy chromatin nucleus usually round Life span = years |
lymphocyte
when active they are larger |
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what is specific about neutrophil chromatin?
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coarse dense
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Lymphocyte ranges:
relative absolute (children) 12months 6years |
Relative 13-40 %
Adult Absolute 1000-4800/uL Pediatric 12 months 4000-10,500/uL 6 years 1500-7000 /uL |
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Abundant, gray-blue cytoplasm, fine ground glass granules
Nucleus is round or kidney bean shaped,brain-like convolutions Chromatin is open “lacy” has vacuoles in cytoplasm |
monocyte
|
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monocyte ref ranges:
relative absolute |
Relative 2-11%
Absolute 200-800 cells/uL |
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Color is pH dependent with Wright stain
Large red/orange granules contain antihistamine Increases are found with parasitic infections and allergic reactions |
Eosinophils
|
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eosinophils are found when and what do their granules contain?
|
Large red/orange granules contain antihistamine
Increases are found with parasitic infections and allergic reactions |
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eosinophil ref ranges:
rel abs |
Relative 0-3%
Absolute 0-500 cells/uL |
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dense, segmented nucleus
large, coarse, blue-black granules cytoplasm may be degranulated granules contain heparin, histamine |
basophil
|
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basophil ref ranges
rel abs |
Relative 0-2%
Absolute 0-200 cells/uL |
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platelet ref range:
abs |
150-450 K/uL
|
|
what is platelet satellitism
|
Platelets are functionally potent and react with a variety of protein receptors and negatively charged surfaces, such as glass,
collagen, basement membrane |
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Both rely on cells settling and measuring red cell mass
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hematocrits and ESRs
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microhematocrit
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volume < 0.1ml
Spun crit Capillary collection acceptable |
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macrohematocrit
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volume 1.0 ml
ESR Capillary collection unacceptable |
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hematocrit give you direct and indirect measurements of what?
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Direct measurement of red cell mass
Indirect measurement of oxygen carrying potential of the blood |
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Volume of packed red cells/volume of WB
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hematocrit
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Expressed as L/L or % of WB
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hematocrit
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Hct is evaluated to assess _____
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anemia
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refers to a low volume of packed red cells/whole blood volume
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anemia
|
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Acute loss of red blood cells
Chronic loss of red blood cells Inability to make red blood cells all cause what? |
anemia
|
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when do you decide on a transfusion?
what is a critical call value for HCT |
when HCT < 30%
<18% |
|
HCT can tell you 4 things
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transfusion need
renal function: Erythropoiesis Function of EPO stimulating red cell production determines blood loss dehydration: HCT is elevated Loss of tissue fluids (plasma) concentrates red cell mass |
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expressed as percent
|
hematocrit
|
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if 15 g/dL is the hemoglobin value then what is the Hematocrit value
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For Hct = HGBx3 +/- 3
so for 15g/dL Hgb 15x3 +/- 3 = 45% +/- 3 You would expect a 15g/dL Hb= 42%-48% Hct |
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what is the normal % for hematocrit
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45%
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how is HCT calculated?
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Hct = MCV x RBC divided by 10
MCV is measured directly as the individual cell size and then a mean is calculated for the population of red cells |
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what are sources of error for manual HCT
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Centrifugation
Maximum cell pack requires correct centrifuge speed 1-4% plasma is trapped in the packed red cells giving a falsely elevated manual HCT reading Misreading of the red cell, clay or plasma interfaces Insufficient mixing of WB specimen before sampling |
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what are sources of error for calculated HCT
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Anything that alters the red cell size
Hyperglycemia – cells swell Hypernatremia – cells swell WBC counts > 35,000/uL affect the MCV Coincidence counting Cell size alters in storage Insufficient mixing of whole blood alters total red cell count |
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what is hyperglycemia and what does it do to RBC
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high glucose content in WB cause RBC to swell
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hypernaremia
what is it and what does it do to RBC? |
high sodium in blood and causes RBC to swell
|
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what can cause error in bother calculated and manual HCT
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Short draw has a dilutional effect with liquid anticoagulants
Incorrect anticoagulant – heparin crenates (spiky) RBCs upon standing IV fluid from central lines may dilute sample Tissue fluids alter according to time of day Patient hydration/IV status Hemolysis – destroys red cell mass |
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top 5 lab errors with HCT
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Improper mixing #1 lab error
Age of specimen Improper reading Problems with high WBC count -Buffy coat |
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Adult reference range for HCT AND HGB
Males and females |
Adult reference range
Males 41 - 53%Hct Hgb 13-17 g/dL Females 36-46% Hct Hgb 12-15.2 g/dL |
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REF RANGES FOR HCT AND HGB
BIRTH 8WEEKS 6MONTHS 12MONTHS 6 YEARS |
Birth 63% Hgb ~20 g/dL
8 weeks 31% Hgb~ 10g/dL 6 months 36% Hgb ~12g/dL 12 months 35% Hgb ~12 g/dL 6 years 38% Hgb ~13 g/dL |
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HCT DECREASE AFTER 65 YEARS OLD WHY?
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BM becomes less productive
70% adipose tissue |
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what does high altitudes do to HCT
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increases
|
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pregnancy _____ hct.... explain why?
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decreases
Increase in red cell production Greater increases in tissue fluids Net decrease in red cell mass/total blood volume |
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smokers hct is ________ explain why
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increased
Carbon monoxide binds to Hgb so that O2 cannot Therefore, more red cells must be produced to carry the same amount of O2 |
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newborns have an ______ HCT why?
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increased
Intrauterine life is hypoxic |
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what is ESR? what is it measured in and how is it measured?
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Rate at which cellular elements settle out of a sample of whole blood – no centrifugation
measured in mm distance fallen in 60 min |
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What is the Principle of the ESR... how does it work?
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RBCs have a negative charge
-Sialic acid residues -RBCs repel each other -Zeta potential Asymmetric plasma proteins decrease negative charge, thus decrease zeta potential As zeta potential is overcome, cells stack together Cells stack in Rouleaux Density overcomes surface area Clumps of cells fall |
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what are the phases of ESR... in mins
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10 min Rouleaux formation takes place
40 min sedimentation phase 10 min packing phase |
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agglutination happens due to what?
|
antigen antibody interaction
|
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what do increases in ESR indicate?
is esr a good screening tool? |
Increases indicate an active inflammatory process is occurring
yes |
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Hyperfibrinogenemia does what to ESR and is caused by what?
|
– increases ESR
Tissue necrosis Infection Pregnancy |
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Haptoglobin does what to ESR and is caused by what?
|
Haptoglobin increases ESR
increase Chronic infection increase Malignancy |
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what is CRP
|
c reactive protein factor in ESR
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High RBC counts/plasma volume result in less plasma and therefore a net ____ in proteins present to reduce zeta potential
|
decrease
|
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Biconcave discs Rouleaux best
because ____ and _____ are slower at compacting... |
Spherical cells are slower
Sickle cells are slower |
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what are some sources of error with ESR
|
Tube length and bore diameter
Tube position Tilt from vertical 3% tilt causes a 30% change in result Temperature -Use ambient -Refrigerated, stored samples have increased viscosity -Samples must come to room temperature before setting up Surface vibrations Centrifuge on counter with sed rate rack Refrigerator motor Hemolysis Bubbles don’t allow cells to settle out |
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the older the patient the _____ in the ESR
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increase
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with ESR what anticoags should be used
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EDTA
Na citrate Oxalate shrinks cells Heparin alters zeta potential |
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ESR is _______ in females
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higher
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westergren, wintrobe and mini ves are all what?
|
ESR tests
|
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westergren
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EXTRA DILUTION STEP and we used in lab
Tube: 230mm x 2.65mm 180 gradations Sample: 1.0 ml of diluted blood Time 1 hour Normal ranges 0-15 mm/hr male < 50yrs 0-20 mm/hr female < 50 yrs 0-10 mm/hr children > 65 years increased normal |
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wintrobe
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Tube: 115mm x 3.0 mm
100 gradations Sample: 1.0 ml WB Time 1 hour Normal ranges 0-15 mm/hr males 0-25 mm/hr females |
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mini ves
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Velocita Erthrocyte Sedimentation
Semi-automated method decreases time to settle Results read in 20 minutes Tube deliberately set at 18 degree tilt |
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what is good about westergren and what is is closely compared to?
|
Westergren uses a larger scale
More sensitive to slight changes Ideal for following changes in rheumatological disease monitoring Decreased, or normal ESR indicates medication is working or indicates disease remission Mini-VES compares closely to Westergren ESR |
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buffers blood by controling pH and is ____ the RBC volume and ____% the RBC weight, and is a protein
|
hemoglobin
1/3 90% |
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Both the _______ and ______keep Hgb functional
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RBC membrane and metabolic pathways
|
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three things that could effect Hgb structure
|
Hb polymerization
Hb crystallization Hb denaturization |
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describe hemoglobin structure
|
Hemoglobin is a tetramere
Heme + globin 4 units /hgb molecule; two dimer protein pairs A heme unit is attached to each globin chain Oxygen attaches to the heme unit |
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Each globin chain is a ____structure of folded segments of amino acids.
|
tertiary
|
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what are the adult Hb Dimers
|
alpha2beta2 HbA1 97%
alpha2 gamma2 Hb F 1% alpha2 delta2 HbA2 2.5% |
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when would you see:
Epsilon chains Zeta chains Hb Gower I Gamma chains Hb Portland |
first 20 days - yolk sac
|
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when would you see:
Gamma chains Alpha chains HbF |
6 weeks gestation- fetal liver
|
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when would you see:
Alpha chains Beta chains HbA1 |
20 weeks gestation -fetal BM
|
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when would you see:
Alpha with Beta-Gamma-Delta chains |
birth- BM
|
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what chain does HBF have and where is it produced?
|
fetal hemoglobin
predominant in fetus made in liver and spleen 2alpha and 2 gamma chains |
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___ comprises 90 -95% of HGB in fetus
|
HBF
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____ has higher affinity for oxygen then HB A1
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HBF
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what is HGB - A1 composed of and who has it>
|
adults and alpha 2 and beta 2 chains
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Beta chain synthesis does not exceed gamma chain synthesis until _____
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after birth
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Following birth, _____ will increase for the next 12 months
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HB A
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a small amt of _____ is produced after birth
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HBF
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when is HGB A2 found and what chains is it made of?
|
alpha and delta....
Occurs late in fetal life Delta chain production begins 7.5-8 months gestation 2.5% found after birth |
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what is HGB is found in babies at birth
|
HBF - 70%
HGB A1 30% |
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what hgb is found in adults
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Hb F - 1%
Hb A1 - 97% Hb A2 – 2.5% |
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Once the red cell nucleus (DNA) is removed, no additional _________is made
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hemoglobin
|
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____% of hemoglobinization is complete while the nucleus is intact
____% of Hgb is assembled after the nucleus is extruded |
65% of hemoglobinization is complete while the nucleus is intact
35% of Hgb is assembled after the nucleus is extruded |
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how is HGB still made after nucleus extruded
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Assembly of chains occur while still attached to the ribosome
|
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Subunits combine in the ______, pyrrole ring insertion occurs in the ________
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Subunits combine in the cytoplasm, pyrrole ring insertion occurs in the mitochrondria
|
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a porphyrin ring + ferrous iron (Fe+2) atom.
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heme
|
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heme is made of what?
|
a porphyrin ring + ferrous iron (Fe+2) atom.
|
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Heme's pyrrole ring sits in a _________pocket of the globin chain, near the surface.
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hydrophobic
|
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Iron can only bind oxygen in the ________ state.
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ferrous
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Once the heme unit is complete, it goes into the cytoplasm to combine with the__________
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globin chains
|
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heme synthesis occurs where?
|
mitch and cytoplasm
|
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ferrous iron is the ____ form
Ferric iron is the _____ form... which can bind to oxygen? |
reduced
oxidized Ferrous |
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what is the rate limiting step in HEME synthesis>
|
Glycine + Succinyl Coenzyme A condensate to for 5 delta aminolevulinic acid
|
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where does the rate limiting step happen>
|
mitch
|
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where does heme synthesis begin?
|
begins in mitch-cyto-mitch(iron insertion)-
then heme moves to cyto for globin chains |
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protoporphyrin IX with iron in center is what? where does this occur?
|
heme
|
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Hemoglobin ref range:
male and female |
13-17 g/dl male
12-15 g/dl female |
|
Hgb] is regulated by the control of _________
|
Hgb] is regulated by the control of erythropoiesis
|
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Patients have shortness of breath, headache, lethargy, dizziness, increased heart rate
sign of what and what is there HGB |
anemia
Hb < 10 g/dL |
|
anemia is associated with what HGB
|
Hb < 10 g/dL
|
|
mild loss of red cell 20% 12g/dL
mod loss of red cells 30-40% 10g/d severe loss of red cells> 50% 7g/dL what do each lead to? |
SOB
SHOCK DEATH |
|
Renal patients have Hb = 7-10 g/dL range
Chronic GI bleeders have Hb = 10 g/dL Slow blood loss patients are admitted to ER with Hb < 4 g/dL HOW IS THIS POSSIBLE? |
Adaptations to red cell loss permit life below a 50% decrease in hematocrit/hgb (red cell mass)
|
|
HGB conc depend on :
|
age gender diet
|
|
HGB RANGES:
g/dL g/L Newborns: Children: Male : Female: Critical call: Critical call: |
g/dL g/L
Newborns: 16-23 160-230 Children: 10-14 100-140 Male : 13-17 130-170 Female: 12-15 120-150 Critical call: < 6.6 < 66 Critical call: >19.9 >199 |
|
Conditions of increased red cell need:
|
high altitude
emphysema cigarette smokers long distance runners pregnancy |
|
Measured Hb < 10 g/dL
physiological need for more O2 associated with what? |
anemia
|
|
with anemia what are some immediate physiological responses?
|
Increased heart rate
Increased respirations Increased circulation |
|
with anema what are some Slower physiological response--long term adaptations
|
Varying environmental conditions or physiologic demand for O2 changes the function of Hb and directly affects the release of oxygen. Physiological changes increase O2 utilization by the body.
|
|
shifting the Hb-O2 curve to the right does what?
|
allows HGB to give up O2 more easily
|
|
what would give the HGB - O2 curve a right shift?
pH pO2 temp pCO2 2,3 DPG |
dec pH lactic acid build up
dec pO2 lack of oxygen in the tissues inc temp higher body temperature inc pCO2 build up of carbon dioxide inc 2,3 DPG places Hgb tetramere in “stressed” configuration |
|
Hb-O2 ~ __% saturated at normal pH in the tissues
Hb-O2 ~ __% saturated at acid pH in the tissues |
50%
25% |
|
what would cause a left shift?
pH pCO2 2,3 DPG temp p02 |
Inc pH (dec H+)
Dec pCO2 Dec 2,3DPG Dec temp Inc in pO2 |
|
_______is produced from the glycolytic pathway.
|
2,3 DPG
|
|
what does 2,3 DPG do?
|
When inserted into the heme unit, Hb bonds are stressed, placing the pyrrole ring at the surface.
O2 is more easily delivered to the tissues |
|
the bohr effect:
As [H+] builds up in the tissues, O2 is more easily released from Hgb. Excess [H+] is then bound to the ____________ |
deoxyhemoglobin
|
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The _________accounts for 70% of CO2 transport to the lungs. This is an_____ transport.
|
The chloride shift accounts for 70% of CO2 transport to the lungs. This is an indirect transport.
|
|
___% of CO2 is directly bound to Hb and transported.
|
25%
|
|
_____% of CO2 is dissolved and carried by the plasma
|
5%
|
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when measuring hemoglobin what are the steps....
|
Whole blood is added to modified Drabkin’s reagent [K3Fe(CN)6].
The red cell is lysed; Hgb is released. All Hgb is converted to the ferric state (Fe+3) of methgb. MetHgb is converted to cyanmethemoglobin by KCN. Absorbance of Hb is measured at 540 nm. Concentrations are read off the standard curve. |
|
true or false standard curve is not prepared for each lot of reagent
|
false
|
|
controls of _____ values are run along with each patient
|
known
|
|
what is done special with patients of unknown hemoglobin values.....
|
run duplicate
|
|
what do you all put on the Hemoglobin standard curve?
|
Your name, date, lot # of std
Title graph: Hgb Std curve X&Y axis; Abs and Hgb conc. Place abs readings on back. Place any comments on back. |
|
dilute _____ g/dL standard to make a curve
|
20
|
|
how do you find the conc of the unknown without a curve
|
Conc unk = (conc of stand x abs of unk )/ abs of stand
|
|
Oxyhemoglobin –
Deoxyhemoglobin – |
Oxyhemoglobin – O2 attached
Deoxyhemoglobin – O2 released |
|
Methemoglobin –
|
Methemoglobin – Heme iron is oxidized (Fe+3
|
|
who is more like to have METHEMOGLOBIN and why?
|
Infants are more susceptible
-HbF is more readily converted to MetHb -Infant red cells lack enzymes to reduce Fe+3 |
|
what are signs of methemoglobin and what percentage will cause cyanosis and what percentage will cause death.
|
Blue color to lips and nails
10% methgb = cyanosis 60% methgb = death (hypoxia) |
|
hypoxia
|
without oxygen
|
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Sulfhemoglobin
|
Sulfhemoglobin – sulfur permanently bound to heme
|
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Carboxyhemoglobin
|
CO bound to heme
|
|
what are signs of carboxyhemoglobim?
what is the affinity difference from regular O2 affinity? what would happen if acute? what do smokers have? |
Lips and nails have a cherry red color
CO:heme binding is 200% stronger than 02:heme Acute carboxyhemoglobinemia = death Smokers have increased CO levels |
|
types of hemoglobin to be measured....
|
Oxyhemoglobin
Deoxyhemoglobin Carboxyhemoglobin Methemoglobin Sulfhemoglobin |
|
what hemoglobin cant be measured with the cyanmethemoglobin?
|
sulfhemoglobin
|
|
which hemoglobin requires a longer reaction time to oxidize the hemoglobin to methgb.
|
carboxyhemoglobin
|
|
hemoglobin is recycled.... it breaks down into _____ and _____. where does they iron go?
|
heme(iron) and globin
|
|
rbc life span is ____ and then senescent cells are what?
|
120 days
engulfed by macrophages or they degrade extra or intervascullary..... |
|
which is good
extravascular degradation or intravascular degradation of hemoglobin/rbc? |
extravascular since heme is toxic when intravascular....
|
|
extravascular degradation
|
hemoglobin catabolized outside of the vascular space (RES cells);
normal mode of recycling |
|
intravascular degradation
|
red cell is lysed intravascularly;
heme, free iron are released into blood stream |
|
Dec haptoglobin levels
Inc urine Hb Inc methemalbumin; Positive Schumm’s Test are all signs of what? |
intravascular degradation
|
|
Inc serum bilirubin
Inc urine urobilinogen Inc fecal urobilinogen Dec H&H levels are all signs of what? |
extravascular degradation
|
|
what are signs of extravascular degradation
|
Extravascular
Inc serum bilirubin Inc urine urobilinogen Inc fecal urobilinogen Dec H&H levels |
|
what are signs of intravascular degradation?
|
Dec haptoglobin levels
Inc urine Hb Inc methemalbumin; Positive Schumm’s Test |
|
MCV-
MCH- MCHC - RDW- |
MCV- mean corpuscular volume
MCH- mean corpuscular hemoglobin MCHC - mean corpuscular hgb concentration RDW- (average) red cell distribution width |
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true or false MCV is not patient specific
|
false
|
|
changes in MCV may indicate what?
|
lab error since it is patient specific
|
|
_____ help categorize anemias and indicate iron storage levels
|
Indices
|
|
what is the MCV equation and what are the units?
|
(HCt/ RBC)x10
femtoliters |
|
what does MCV tell you?
|
red cell size
|
|
MCV ref ranges:
normocytic microcytic macrocytic |
normocytic 80-100 fL
microcytic < 80 fL macrocytic > 100 fL |
|
what causes macrocytosis?
|
Young blood cells (shift cells)
Newborns Post surgical Post partum Pregnancy Iron replacement therapy Hemolytic anemia HDFN B12 or Folate deficiency Alcoholism Liver disease Chemotherapy |
|
what causes microcytosis?
|
Iron deficiency
Thalassemia Decreased iron utilization RBC fragmentation |
|
MCH tells you what and what is the equation, and what is the units?
|
hemoglobin content
(HGB/RBC) x 10 picograms |
|
not sex dependent
related to HGB within a red cell |
MCH
|
|
ref range for MCH
|
26-34pg
|
|
what are the times three rules?
|
Hb X 3 = HCT
RBC X 3 = Hb |
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MCH does not consider overall ___________, so it not as useful as the MCHC for QA
|
MCH does not consider overall cell size, so it not as useful as the MCHC for QA
|
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what does MCHC tell you... what is the equation and what are the units
|
color of red cell
(HGB/HCT) X 100 grams/dL % |
|
Reference Ranges for MCHC
Normochromic Hypochromic Hyperchromic |
Reference Ranges (color)
Normochromic 32%-36% Hypochromic < 32% Hyperchromic > 36%* |
|
Tightest RBC parameter
Best index for QA |
MCHC
|
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Markedly increased with RBC agglutination or spherocytes
|
MCHC
|
|
MCHC > 36 typically indicates
|
analytical error and RARELY spherocytes
|
|
RDW what is the equation and units?
|
Units = (%)
(SDt/MCV) x 100 |
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RWD ref range:
|
11.5-14.5%
|
|
RDW > 16 indicates ?
|
anisocytosis
|
|
individual size difference of RBC
|
RDW
|
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The bigger the RDW, the _____ size variance you will see under the microscope.
|
more
|
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indices are used when reporting what?
|
Descriptors of red cell size and color are reported when the patient has anemia
|
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closeness of the analytical result to the true value
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accuracy
|
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the ability to reproduce the result
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precision
|
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preanalytical error
|
any error that occurs before the sample is analyzed
Specimen problems |
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systematic error
|
analytical error that occurs because of variances in the procedure
Instrument problems Reagent problems QC material with known values ID systematic error |
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radom error
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sporadic systematic error that occurs without an identifiable source
Duplicate testing eliminates random error |
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post analytical error
|
errors that occur during the reporting phase of laboratory testing.
Data utilization errors TAT too long Reported result uses incorrect units |
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______ provide inside into the expected red cell morphology
|
red cell indices
|
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mean cell volume measured in what?
|
fL
|
|
hematocrit units
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%
|
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mean cell hemoglobin units
|
pg
|
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mchc units
|
g/dl %
|
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rdw units
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%
|
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diffusely basophilic erythrocytes also known as
|
reticulocytes
|
|
reticulocytes describe color and properties
|
immature RBC
staing bluish and slightly bigger than RBC carry less O2 since not fully hemoglobinized and hemoglobin assembly still takes place due to lots of ribosomes even though nucleus may be gone. you dont want to see in a PBS |
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anisocytosis
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variation in RBC size
|
|
Microcytosis causes:
|
iron def
thalassemia sideroblastic anemia |
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macrocytosis causes:
|
megabloblastic anemia
hemolytic anemia hemorrhage liver disease asplenia endocrinopathies |
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poikilocytosis
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alterations in red cell shape
|
|
burr cells formal name
|
echinocytes
|
|
what do echinocytes look like
|
evenly distributed pointy projections, crenated
|
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why do echinocytes form?
|
increased pH (from slide)
septiciema renal insufficiency artifactual |
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why are echinocytes shaped the way they are?
|
outer layer has more lipids giving it a rough shape
|
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mouth cells formal name
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stomatocytes
|
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uniconcave cells, increased cell membrane surface are. inner inner layer has more lipids than outer
|
stomatocytes
|
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what causes stomatocyte formation
|
alchoholism
liver disease |
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these can be genetic of acquired .... no central pallor and have a MCHC over 36%
|
spherocytes
|
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rbc fragments due to mechanical damage are called
|
schistocytes
|
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true of false... schistocytes can still deliver oxygen
|
true
|
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what are schistocytes caused by...
|
fibrin strands
severe burns mechanical heart valves glomerular nephritis |
|
hemolysis makes cells ____ so there are no _____
|
explode
fragments |
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formal name of spur cells
|
acanthocytes
|
|
describe acanthocytes
|
thorn like spikes
and they are irregular bublbous projections |
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what causes acanthocytes?
|
changes in plasma lipids
alchoholic cirrhosis, lipid metabolism disorders |
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formal name for target cells
|
codocytes also called bulls eye or mexican hat
|
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what causes codocytes
|
increase cholestrol and lipids
|
|
the skin is to lose for the body... increased surface area to volume ratio
|
codocytes
|
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rbc fragments due to mechanical damage are called
|
schistocytes
|
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true of false... schistocytes can still deliver oxygen
|
true
|
|
what are schistocytes caused by...
|
fibrin strands
severe burns mechanical heart valves glomerular nephritis |
|
hemolysis makes cells ____ so there are no _____
|
explode
fragments |
|
formal name of spur cells
|
acanthocytes
|
|
describe acanthocytes
|
thorn like spikes
and they are irregular bublbous projections |
|
what causes acanthocytes?
|
changes in plasma lipids
alchoholic cirrhosis, lipid metabolism disorders |
|
formal name for target cells
|
codocytes also called bulls eye or mexican hat
|
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what causes codocytes
|
increase cholestrol and lipids
|
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the skin is to lose for the body... increased surface area to volume ratio
|
codocytes
|
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what environmental factor can cause target cells?
|
high humidity
|
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what is the formal name for tear drop cells
|
dacryocytes
|
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why are dacryocytes formed?
|
inclusion are removed by spleen
|
|
what caues dacryocyte formation
|
myelodysplastic syndromes
|
|
another name for sickle cells
|
drepanocytes
|
|
genetic abnormality that result in HbS formation
|
sickle cell disease
drepanocytes |
|
HbS tactoids and polymerization of Hb
|
sickle cell disease
drepanocytes |
|
what causes sickle cells to unsickle
|
reoxygenation but can only do it a few cycles
|
|
what would Na metabisulfite be used for?
|
to test for sickle cell disease
When a fresh solution of sodium metabisulfite (reducing soln) is used, positive cases should show 10 to 75 percent sickling within 15 minutes. There is a positive correlation between the degree of sickling and the severity of the disease. (3) Normal blood treated in this way shows no sickling. |
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formal name for cigar/pencil cells
|
elliptocytes or ovalocytes
|
|
what are elliptocytes/ovalocytes caused by?
|
hereditary membrane abnormality or can be acquired artifactual
|
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formal name for helmet cells
|
keratocytes
|
|
another name for blister cells
|
keratocytes of helmet cells
|
|
hornlike projections cells
|
keratocytes
|
|
what causes keratocytes
|
cause by removing LARGE inclusions by spleen
or cells care caught upon intravascular fibrins strands |
|
HbC crystals
|
Hemoglobin C comprises 2 normal alpha chains and 2 variant beta chains in which lysine has replaced glutamic acid at position 6. This unstable hemoglobin precipitates in red blood cells to form crystals. These intracellular crystals lead to a decrease in red blood cell deformability and an increase in the viscosity of the blood. The spleen effectively removes these crystal-containing cells.
Much like the mechanism in sickle cell hemoglobin, the amino acid change in the hemoglobin C molecule impairs malaria growth and development. It reduces parasitemia and confers protection against mild malaria attack.1, 2 Therefore, persons who are heterozygous for hemoglobin C have a survival advantage in endemic areas. The risk of malaria is lower still in persons who are homozygous for hemoglobin C. |
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when would crystals and lots of target cells be seen?
|
when a patient has inherited Hb C
few target cells seen in Hb AC which is a carrier state |
|
Hbc is acquired how? and what gets rid of them?
|
genetically
and are removed by spleen |
|
Polychromasia
|
prescence of reticulocytes normal to a certain extent
A slight degree of polychromasia is normal. Increases in polychromasia suggest an increased bone marrow response. No polychromasia correlates with non-responsive, anemic patients, which is characterized by all cells exhibiting the same coloration. This finding presents a guarded prognosis, pending resolution of the cause of the non-regeneration. |
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which cell types follow these guidelines?
occ: 0-2 moderate: 2-5 many: >5 |
stippling
howell jollu bodies pappenheimer bodies spherocytes tear drops ovalomacrocytes |
|
which cells follow these RBC quidelines?
occ: 2-5 moderate:5-20 many: >20 |
burr cells
elliptocytes target cells |
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which RBC types follow these guidelines?
occ: 2-3 moderate:4-6 many: >6 |
polychromasia
|
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which RBC types follow these quidelines:
occ:5-20% moderate:20-40% many: >40% |
macro/microcytes
|
|
what is dyserythropoiesis?
|
when rbc production goes wrong and nucleus hasnt been fully removed
|
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what is left in howell jolly body cells and why?
|
DNA fragment....
from dyserythropoiesis |
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what is a cause for howell jolly bodies?
|
Common causes of asplenia are splenectomy due to trauma, and autosplenectomy caused by sickle cell anemia.
|
|
cabot rings are what and are caused by what?
|
Cabot rings are thin, red-violet staining, threadlike strands in the shape of a loop or figure-8 that are found on rare occasions in erythrocytes. They are believed to be microtubules that are remnants from a mitotic spindle.
Cabot rings have been observed in a handful of cases in patients with megaloblastic anemia, lead poisoning and other disorders of erythropoiesis. such as dyserythropoiesis |
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what causes Heinz Bodies?
|
denatured Hb
|
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how can you see Heinz Bodies?
|
supervital stains
|
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how many heinz bodies per cell?
|
one
|
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how many howell jolly bodies inclusions are in a cell?
|
few
|
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pappenheimer bodies have what inclusions?
_______ deposits _______ granules |
iron deposits
siderotic granules |
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what are siderotic granules
|
iron deposits
|
|
how many pappenheimer bodies inclusions are seen in cells?
|
many
|
|
what are the two intracellular parasites we talked about
|
malaria - africa
babesia - USA |
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how is malaria and babesia acquired?
|
malaria - misquito
babesia- tick |
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what would you see with intracellular parasites?
|
ring froms with dark spot inside red blood cell
|
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what stains can nucleated RBC be seen in and what is the inclusion
|
wright and supervital
dna |
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what stains can reticulocyte be seen in amd what is the inclusion?
|
supervital only
rna ribosomes |
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what stains can basophilic stippling being seen in and what is the inclusion?
|
can be seen with both the supervital and wright stain
the are RNA |
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what is basophilic stippling and what is it caused by?
|
rna inclusions
black to dark purple stuff with in RBC caused by lead poisoning |
|
with what stain can howell jolly bodies be seen with and what is the inclusion made of?
|
supervital and wright
DNA |
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with what stains can Heinz bodies be seen and what is the inclusion made of?
|
supervital only
hgb |
|
what two cell type inclusions can only be seen with supervital stain?
|
reticulocytes and heinz bodies
|
|
with what stains are pappenheimer bodies seen and what are the inclusions made of?
|
supervital and wright
iron siderotic granules |
|
what results from inc plasma proteins.....(rbc)
|
rouleaux
|
|
what disease causes rouleaux
|
multiple myeloma
|
|
this results from IgM cold Ab
|
agglutination
|
|
manual counts are counted in ___ ______ volumes
|
one microliter
|
|
to convert one microliter to one liter what do you multiply by?
|
10^6
5.5 x 10^3 = 5.5 x 10^9 cells/L |
|
Normal WBC count
|
4500 - 11500 cells/microliter
|
|
how should WBC counts be reported?
|
##.# x 10^3/microliter
##.# x 10^9/liter sometimes ##.# thou/microliter |
|
what kind of blood should be used for manual cell counts
|
edta whole blood
body fluid samples |
|
Manual counting procedure requires three things....
|
1. diluting fluid
2. dilutiong pipette 3. hemacytometer |
|
what diluting fluids would you want to use when looking for WBC?
|
turks
1% HCl 2% glacial acetic acid AMMONIUM OXALATE -lyses RBC |
|
ammonium oxalate is used when?
|
diluting fluid used to lyse rbcs and read WBC or PLATELET counts
|
|
what diluting fluids would you use to read RBC counts
|
saline -reduces plasma proteins and stabalizes rbc
|
|
with the unopette system what dilutions do you wanna make for
RBC WBC PLATLET and with what? |
for RBC you wanna make a 1:200 dilution with saline
for platelet and WBC you wanna make a 1:100 dilution with ammonium oxalate |
|
how do you calculate # of cells counted?
|
# cells counted/microliter = [(avg # of cells)x dilution]/(area x depth)
|
|
for unopette WBC count what is the equation?
|
(avg cells x 100)/(9mm^2 x.1mm)
|
|
what to do in high # of cells during manual count
|
increase dilution
|
|
what to do in low # of cells during manual count?
|
use larger volume
smaller dilution count more areas |
|
Neubauer Chamber
|
H chamber
used for manual counts groud glass precision instrument depth is constant two counting sides |
|
for WBC counting how many of the squares do you count?
|
you count all the 9 lrg squares which are 1x1mm each
|
|
for rbc counting what squares do you count?
|
5 small squares within large center square
|
|
for platelet counting what squares do you count?
|
all 25 small squares within lrg center square
|
|
other Hemocytometers
FUCHS ROSENTHAL SPEIRDS-LEVY |
FR: 4mmx4mm depth=.1mm volume=1.6mm^3
SL: 5mmx2mm depth=.2mm volume=1.6mm^3 |
|
Cell counting procedure:
8 steps |
1. make dilution
2. clean chamber 3. set coverslip 4. charge both sides 5. let PLT counts sit in humidity chamber while they settle to avoid dehydration 6.count cells that fall at top and left line 7.exclude right bottom line cells 8. count both side and avg within 10% |
|
with cell counting what must the avg of both sides must be within what percent?
|
10%
|
|
sources of error with Manual counting
|
1.poorly mixed sample
2.not whipping off uniopettes 3.overzealous mixing and spilling dilutant 4.overfilling chamber 5.evaporation 6.uneven cell distribution due to ling or uneven charge 7.not letting PLT settle 8.using too much light 9.incorrecting identify cells 10.not using formula correctly |
|
what does overfilling a chamber do?
|
falsely lowers cell count
-excess fluid flows into moat pulling cell particles out of counting area |
|
what does evaporation do to a manual count?
|
falsely increases cell count
-large cell # takes to long to count -fluid evaporates w/ light intensity |
|
when doing manual count what must you do with the light...
|
low light and lower diaphragm
|
|
Coulter principle with counting....
|
impedence couting uses interruptions in an electrical current to detect cell particles
-voltage spikes = # of cells counted -larger spikes (cell volume) is proportional to size of resistance -relative frequency of occurence is plotted in a histogram plot of MCV |
|
COULTER AcT
-sample size -WBC and RBC dilutions done together or seperate? -cell counts repeated how many times? -what is the HGB measured from? |
100 microliters of whole EDTA blood
seperate dilutions cell counts done in triplicate hgb measured from WBC dilution |
|
Laser technology how does it work with cell counting
|
the way the cells flow through and the light scatters lets you know different characteristics of a cell
|
|
RALS
FALS neutrophil would have what? |
RALS measures how complex the interior is
FALS measures size high fals and high rals |
|
what would be Coulter machine errors?
|
1. poor mixing
2. fibrin/platelet clots 3. instrument error 4. patient specimen interferences -lipemia -cold agglutinins -difficult draw -short draw -IV contamination |
|
what is lipemia?
|
cloudy fatty plasma
|
|
what kind of counts can be done on peritoneal(dialysis fluid) and ventricular(brain ventricular space) fluid types?
|
RBC, WBC, Bacteria
|
|
what kind of counts can be done on Thoracic(upper chest) and Pleural(lung) fluid types
|
RBC, WBC, tumor cells
|
|
what kind of counts can be done on cerebrospinal fluid?
|
RBC, WBC, bacteria, tumor cells
|
|
what kind of counts can be done on Synovial joint fluid types
|
rbc,wbc,bacteria,crystals
|
|
what kinda of counts can be done on pericardual(heart lining) fluid counts
|
WBC, tumor counts
|
|
how are the following labeled:
WBC RBC HGB HCT MCV MCH MCHC PLT |
WBC- #.# x 10^9/L or ####/microliter
RBC- #.## x 10^12/L HGB- ##.#g/dL HCT- ##% MCV- ## fL MCH- ## picograms pg MCHC- ## % PLT- ### x 10^9/L |
|
Rel cell type
absolute cell type what are the units? |
relative cell ##%
absolute cell #,###/microliters |
|
what does the Hemocue measure?
|
HGB
|
|
what is an advantage of the hemocue over the coulter
|
smaller is to travel with and only needs cappillary blood.
much faster as well |
|
when make a cyanmethemoglobin curve what is labeld on the axis's
|
Hemoglobin g/dL on X
Absorbance at 540nm on Y |
|
the bohr effect refers to the cells ability of....
|
accepting H+
|
|
a person with renal disease would have
|
decreased hematopoiesis
erythropoeiten is not made and hematopoiesis is decreased. |
|
3 phases of ESR
|
10 min forming rouleaux
40 min falling 10 min stacking |
|
why do plasma proteins increase ESR?
|
the decrease the neg charge thus decreases the zeta potential of RBC
|
|
what is an acceptable agreement btwn a PBS WBC estimate and a Automated WBC count?
|
25%
|
|
and increase in macrocytic, polychromatphillic erythroctytes seen in wright stain smear should make you wanna do what test?
|
reticulocyte count
|
|
what are you lookin for in wright stain lower power
|
color and no precipitates
|
|
with lipemic blood sample which paramters will be affected?
WBC RBC HGB HCT MCV MCHC MCH |
MCHC,MCH,HGB
|
|
allowing slide to dry vertically doesnt allow _______ to form
|
precipitate
|
|
specimen for wedge smear
and stain used |
whole blood with anticoagulant EDTA
wright stain if not stainin immediately let air dry and fix with methanol.. otherwise stain fixes with alcohol |
|
wright stain effect is produced by the combinded action of ________ _____ and its oxidation products ________ and ___________
|
methylene blue
eosin Y and azure B |
|
how to do a wright stain
|
1. place slide smear up on rack
2. flood with methanol, fix for 5 min 3. drain meth and blot with filter paper 4.flood with wright stain for 3 min 5.add wright buffer until greenish metallic sheen comes up... mix by blowing 6.allow mix to stand 8-10 min more wbc more time needed 7.float off green mix with distilled h2o 8.keep smear horizontal 9.blot smear immediately blow dry 10.wipe back of smear with meth. 11. mount coverslip with permount |
|
lymphocytes have what color granules if present?
|
red
|
|
neutrophil granules are what color
|
purple violet
|
|
monocyte granules stain what color
|
pale red, fine dust particles
|
|
what to do on 10X
|
check color
wbc evenly distributed look for RBC agglutination and rouleaux look for immature abnormal cells, platelet clumps or satellitism do WBC estimate |
|
how to perform WBC estimate
units recorded? |
avg wbc counted in 5 fields and multiply by 200....
should agree with automated WBC within 25% ##.# x 10^9/L or ####/microliter |
|
what to do on 100x
|
platelet estimate and eval.
wbc diff nucleated rbc red cell morph |
|
platelet ref ranges
|
rare: <50K/uL
decreased: 50-149K/uL normal: 150-450K/uL increased: >450k/uL |
|
how to correct for WBC count when there are nucleated RBC
|
(WBC x100)/ (nRBC+100 )
|
|
packed cell volume of whole blood
|
hematocrit
|
|
what kind of specimen is used for Hematocrit (microcappilary)
|
whole blood with EDTA or Heparin
|
|
when measuring hematocrits blood collected by finder prick should be collected in a _______ ________. Blood collected from a venipuncture should be test in an _________ ______________.
|
heparinized capp tube
untreated capp tube |
|
when the Coulter gives a WBC reading exceeding __________ a microcappilary method should be preformed to get hematocrit... .whY?
|
35.0 x 10^9/L
because high WBC maye elevate RBC and hematocrit reading.... coulter calc. HCT from rbc and mcv values |
|
specimen for HCT....
|
whole blood in EDTA.... must be half full so cell done shrink and false lowered HCT
or if capp blood avoid edema areas |
|
when doin HCT always make _________ capillary tubes and stopper with _____
|
2, clay
|
|
how long do you spin HCT tubes?
|
5 mins
|
|
when using the reader for HCT what do you do
|
clay/RBC is set to zero
plasma/air is set to 100 HCT is taken at RBC/plasma interface |
|
duplicate microcapp tubes for HCT should agree within what %
|
+/- 3%
|
|
when recording HCT what two things do you record?
|
your duplicates
average as whole number % |
|
ref range for HCT
male female |
male 41-53%
female 36-46% |
|
RBC surface is ________ charged
|
neg
|
|
increased conc of fibrinogen, gamma globulin, and proteins does what to ESR
|
neg charge btwn RBC is reduced, they rouleaux more easily and an increased ESR
|
|
ESR is increased in anemia due to what?
|
easier settling of a small number of cells in a larger volume
|
|
used for the detection and evaluation of the conditions where tissue destruction and inflammation are present
|
ESR
|
|
advantage of using MINI VES analyzer over westergren
|
reduced biohazard exposure
greater accuracy improved precision shorter test only 20 min |
|
what specimen is used for ESR
|
whole blood with EDTA less than 3 hours old if at Room temp
orrr lest the 12 hours old if refrigerated at 4 degree celcius blood tube must be more than half full, free of clots,and at room temp before testing! |
|
how to do ESR
|
1. fill dilution vial to lower line with .9% sodium chloride (.3mL)
2. add well mixed blood to top line (1.2mL) and mix with parafilm cover 3.pipette into westergren pipette until cotton plug is saturated 4.wipe off and place in level Sedirack 5.time for 1 hour 6.red interface of RBC and plasma |
|
how to report ESR
|
report in whole number in mm/hour
|
|
ref ranges for ESR:
Male female children |
Male:
18-50yrs: 0-15mm/hr 50-85yrs: 0-20mm/hr 85 and over: 0-30mm/hr female: 18-50yrs: 0-20mm/hr 50-85yrs: 0-30mm/hr 85yrs and over: 0-42mm/hr children: 0-10mm/hr |
|
ESR sources of error slowing it.
|
bubbles
cold temp increase plasma viscosity dehydration inc plasma viscosity extreme microcytosis excess EDTA high RBC old blood- rbc become spherical and less likely to rouleaux poikilocytes test read too soon |
|
ESR sources of error accelerating it
|
low RBC
high fibrinogen or globulin levels- could be from infalmmation or pregnancy macrocytosis delay in reading surface vibrations tilted tube warm temp |
|
macrocytosis and ESR
|
large cells settle faster than smaller ones
|
|
to measure hemoglobin what method is used
|
cyanomethemoglobin
|
|
what decimal place is Absorbance measured too
|
third
|
|
what is all in DRABKINS reagent
undiluted _____standard is equivalent to _____ |
potassium ferriccyanide
potassium cyanide potassium dihyrdogen phosphate Triton X can be stored for 1month in dark bottle 80mg/dL...... 20g/dL |
|
when making curve what should your dilutions be for
5, 10, 15, 20 and blank g/dL? patient sample are normally diluted what to what how is this prepared with WB? |
5: 1.5mL standard and 4.5mL D.R.
10: 3 and 3 15: 4.5 and 1.5 20: 6 and 0 blank: 0 and 6 1:250 pipette 5mL of Drabkins then add 30uL of blood(wipe tip) and mix well let tube stand for 5 mins zero out specto. and then measure abs |
|
after mixing standard and drabkins how long must you wait?
what must you first do to spectrophotmeter? |
3 minutes
zero with reagent blank |
|
when measuring hemoglobin abs do you start with lowest conc or highest conc?
|
lowest
|
|
a ___________ must accompany each new lot of Drabkins....
|
new standard curve
|
|
principle of drabkins reagent and whole blood interaction
|
WB added to drabkins
RBC lyse and hemoglobin oxidizes to methemoglobin by K ferricyanide during oxidation iron is converted to ferric iron methemoglobin forms with K cyanide to for cyanomethemoglobin which is a stable pigment |
|
what kind of specimen is used during the cyanomethemoglobin method?
|
WB with EDTA or Heparin
or direct capp blood |
|
what are the ref range and units for HGB
male female |
male 13-17 g/dL
female 12-15 g/dL ##.# g/dL |
|
____ is a measurement of the coefficient of variation based on the red cell distribution histogram, rather than a calculation from other red cell parameters
|
RDW red cell distribution width
|
|
RBC morph reporting precedure
10x and 100x |
10x look for rouleaux, agglut., stain quality, platelet clumps at edge
100x look at 10 field min (150-200 cells) RBC should not be touching and avg abnormalites reported (platelets also evaluated) |
|
if seen under low power report as present with RBC
these should be reported if seen at all |
rouleaux or agglut
cabot rings parasites Hgb C crystals |
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how to use unopette system
|
repeat procedure twice and charge each side with one sample
1. puncture reservoir with protective shield 2.fill capp pipette with blood... wipe blood 3.add blood to reservoir 4.cap with shield to prevent evap. 5. let sit 10 -15 min to lyse cells (WBC count should be preformed within three hours) 6.the convert pipette to dropper and charge one side of hemocytometer first view at 10x then at 40x sides should agree with 10% |
|
what do you clean a hemocytometer with?
|
isoterge
distilled h20 and ethanol |
|
what specimen should be use for manual white cell counts?
|
whole blood in EDTA
|
|
what is the dilution for the UNOPETTE
|
1:100
20uL in unopette and 1.98mL of diluting fluid.... |
|
when doing WBC counts and using a 1:20 dilution only count the______
|
4 corner squares
with 1:100 dilution count all 9 squares |
|
cells/ mm^3 is the same as what?
|
cells/uL
|
|
ref range for WBC
|
4.0-11.0 x 10^9/L
|
|
micro
femto pico milli |
micro 10^-6
femto 10^-15 pico 10^-12 milli 10^-3 |
|
ONPG
|
beta-galactosidase
|
|
ONPG results
|
neg colorless
pos yellow |
|
ADH
|
arginine dihydrolase
|
|
ADH results
|
neg yellow
pos red/orange |
|
LDC
|
lysine decarboxylase
|
|
LDC results
|
neg yellow
pos red/orange |
|
ODC
|
ornithine decarboxylase
|
|
ODC results
|
neg yellow
pos red/orange |
|
CIT
|
citrate utilization
|
|
CIT results
|
neg pale green/yellow
pos blue green/ blue |
|
H2S
|
H2S production
|
|
H2S results
|
neg colorless or grey
pos black/thin line |
|
URE
|
urease
|
|
URE results
|
neg yellow
pos red/orange |
|
TDA
|
tryptophane deaminase
|
|
TDA results
|
after adding 1 drop ferric cholride
neg yellow pos brown/red |
|
IND
|
indole production
|
|
IND results
|
after adding one drop KOVACs and waitng two mins
neg yellow pos red ring |
|
VP
|
acetoin production
|
|
VP results
|
after 1 drop K hydroxide and 1 drop alpha naphthol and wait 10 min
neg colorless pos pink/red |
|
GEL
|
gelatinase
|
|
GEL results
|
neg no black pigment diffusion
pos diffusion of black pigment |
|
ALL SUGARS
|
fermentation and oxidation
|
|
ALL SUGARS results
|
neg blue/ blue green
pos yellow |