MTLE refresher notes Flashcards
Hand-held device that aids in visualization prior to phlebotomy
AccuVein
Blood to anticoagulant ratio in black-top tubes
4:1
Blood to anticoagulant ratio in light blue-top tubes
9:1
Anticoagulant used for trace elements and toxicology determination
Sodium heparin
Anticoagulant used for fragility testing and Blood Gas Analysis
Heparin
Disadvantage of using Serum Separator Tubes (SSTs)
It interferes with (TDM) Therapeutic Drug Monitoring
Clotting time for:
Tubes with activator, with gel separators, and without anticoagulant
With gel: 30 mintues
With clot atvator: 5 mins
w/o anticoagulant: 60 mins
arterial puncture order of preference
- Radial artery 2. Brachial artery 3. Femoral artery
Test prior to radial artery puncture
Modified Allen Test
Amount of anticoagulant for arterial puncture
0.05 mL of liquid heparin or 1,000 u/mL
Blood Gas Collection for Newborns
Indwelling Umbilical Artery Catheter
Order of draw for catheter liines
- Discard 2-5 mL of blood and discard.
- Blood culture
- Anticoagulated tubes
- Serum tubes
Order of draw for capillary puncture
- Tubes for blood gas analysis
- Smear/Slide Preparation
- EDTA
- Other anticoagulated tubes
- Serum tubes
Order of Draw for Venipuncture
- Yellow top
- Light Blue top
- Red top (serum tubes)
- Green top
- Lavender top
- Gray top
Size of the drop of blood in wedge smears
2-3 mm
How to make a thin smear
Increased pressure
Decrease the (ASS) Angle, Speed, and Size of blood drop
How to make a thick smear
Derease the pressure
Increase the (ASS) Angle, Speed, and Size of blood drop
Physiological factors that may lead to pre-analytical variations
Position - increase values of large molecules
Diurnal variation- increase (WBCs) eosinophils in the afternoon
Stress - increase WBC count
Smoking - Increase WBC hematocrit, RBC count and hemoglobin
Exercise - increase WBC count
Diet- Affects hemoglobin and coagulation tests
Prolongged tourniquet application results to?
hemoconcentration
Light senstitive substances
Bilirubin, Vitamin A, Beta-carotene, porphyrins, Iron
Samples to be chilled
Ammonia, Gastrin, PTH, Lactic Acid
Blood films from EDTA should be made within how many hours
2-3 hours
EDTA-containg tubes
Lavender
Pink
Tan
White
Royal Blue
Automated smear-making and staining system (wedge type method)
Sysmex SP-10
Automated staining system (dip-batch staining)
MIDAS III
Automated staining (propelled towards two rotating plantars)
HemaTek
RBCs with blunt or pointed protrusions that are evenly spaced
Echinocytes
Macroscopic appearance of a well-stained smear
Purple/Pink-colored film
Macroscopic appearance of a stained film obtained from blood samples with INCREASED PROTEIN LEVELS
Blue color
Macroscopic appearance of stained film obtained from blood samples with INCREASED LIPIDS
Has holes
Macroscoopic appearance of a stained film obtained from blood samples with INCREASED WBC AND PLATELETS
Bluish lateral edges
Macroscopic appearance of a stained film obtained from blood samples with RBC aggregates
Grainy appearance
Objective used to estimate bone marrow cellularity
LPO
Objective used for WBC estimates
HPO
May be confused with plasma cells/plasmablast
Osteoblast
May be confused with megakaryocyte
Osteoclast
Objective used for WBC differential count, platelet estimates, and cell morphology
OIO
RBC per OIO field viewed in the correct are of the film
200-250 RBCs per OIO field
Number off WBCs to be differentiated when WBC count is >40 x 109/L
200-cell differential count
Number of WBCs to be differentiated when WBC count is >100 x 109/L
300 or 400-cell diifferenntial count
Number of WBCs to be differentiated when WBC count is <1 x 109/L
50-cell differential count
Increased number of bands or cells younger than bands in the PBS
Shift to the Left
Is the first immature neutrophil normally found in the peripheral blood
Band/Stab/Staff cell
SIte for bone marow syntesis in adults and children more than 2 yrs old
Posterior Superior Iliac Crest
Site for bone marrow aspirate collection for children less then 2 years old
Tibia
Alternative sites for bone marrow aspirate collection in adults
Sternum
spongy bone spicule of vertebrae
Other red marrow containing sites
Bone marrow sample management
- direct smears
- Anticoagulated smears
- Crush preparation
- Touch print technique
- Concentrated bone aspirate
Used for the evaluation of the morphology and distribution of hematopoietic cells (immature)
Bone marrow aspirate
Used when patients are suspected of focal lesions
Bone marrow core biopsy
Staining technique for bone marrow core biopsy
H & E staining
1 ml of bone marrow in heparin is used in
Cytology studies & Flow Cytometry Techniques
Most common erythrocytic stages found in the normal marrow
Polychromatophilic & Orthochromatophilic normoblasts or erythroblasts
What are the reagents used in Drabkin’s solution
- Potassium ferricyanide - converts hemoglobin to methemoglobin
- Potassium cyanide - converts methemoglobin to cyanmethemoglobin at 540 nm
- Sodium carbonate
- Surfactant
Used to assess the erythropoietic activity of the bone marrow
Reticulocyte count
What is the relationship of ESR to plasma viscosity?
Inversely proportional
What is the relationship of ESR to the RBC mass?
Directly Proportional
Passage of more than one cell a time through the aperture
Coincidence
Measures the pulse generated by the resistance of the cell as it passes through the current within the aperture
Electrical impedance
Measures the rate of conductivity
Radiofrequency
The principle in flow cytometers
Optical scatter & Immunofluorescence
Principle of Optical scatter
Uses laser beam to differentiate between different blood cell types
Principle of Immunofluorescence
Uses immunophenotyping
Uses fluorescent-labeled monoclonal antibodies
Combination of forward-low and forward-high scatter. A principle employed by Siemens system
Differential scatter
Represents excess signals at the lower threshold region of the WBC histogram
R1
Coulter’s technology of five-part WBC differential
VCS technology
MAPSS technology and three-color fluorescence
Abbott Celldyne Sapphire
Peroxidase-staining absorbance and light scatter
Siemens Advia
Detection of forward and side scattered light and fluorescence
Sysmex XN-1000
RBC in TAILS
(Thalassemia, Anemia of Chronic Disease, Iron deficiency anemia, Lead Poisoning, and Sicke Cell disease)
Microcytic
RBC in Vit. B12 deficiency, folate deficiency, and pernicious anemia, Steatorrhea, sprue, D. latum infection
Macrocytic
Test involving the formation of Holly Leaf appearance
Sodium Metabisulfate
Screening test for hemoglobin S solubility
Sodium dithionate
Forward light scatter measures
cell volume
Side-angle scatter measures
internal complexity
Falsely high ESR
Heat from the back of refrigerator
Vibrations from opening and closing the refrigerator
Falsely low ESR
Cool air from refrigerator
Decreased ESR is caused by the what
Poikilocytes
Thalassemia
Over-anticoagulation
Bubbles
Delayed reading
Increased ESR is caused by what
Multiple myeloma
Waldenstrom’s macroglobulinemia
Anemia
Bacterial infection
Carcinoma
Pregnancy
Menstruation
High room temperatures
Tilting
A fluid used for the determination of eosinophil count
Pilot’s fluid
Three components of the Pilot’s fluid
- Phloxine B - stains eosinophil bright red
- Propylene glycol - lyses RBCs
- Heparin - prevents leukocyte aggregation
Diluting fluids for WBC count
Turk’s and HCl
The most commonly used RBC count diluting fluid
Formol citrate
Color of Wright-stained red cells may be adjusted by?
Adjusting the buffer
Causes of excessively pink smears
Thin smears
Acidic buffer
Prolonged washing
Insufficient staining time
Old fixative
Causes of excessively blue smears
Think smears
prolonged staining
Insufficient washing
Alkaline buffer
Heparinized sample
Characterized by increased Osmotic Fragility Testing
Shperocytes
Characterized by decreases OFT
Target cells
Test for the identification of Factor XIII deficiency
Ducker’s test
Type of lymphocyte seen in Infectious Mononucleosis
Type II Reactive Lymphocyte/IM Lymphocyte
What cells are infected by the causative agent of IM
B cells
A reactive lymphocyte also known as “Turk’s Irritation Lymphocyte”
Type I
The causative agent of Infectious Mononucleosis
Epstein-Barr Virus
Mode of transmission of EBV
Kissing
Blood transfusion
Organ transplant
sexual contact
Which cells are infected by EBV
CD21/B cells
A congenital form of Red Cell Aplasia
Diamond-Blackfan Anemia
A congenital form of Aplastic Anemia
Fanconi Syndrome
The classic drug associated with bone marrow aplasia
Chloramphenicol - an antibacterial drug
An inherited form of Aplastic Anemia
Fanconi syndrome and Idiopathic Aplastic anemia
Acquired Aplastic Anemia is caused by what
Chronic exposure to certain drugs and chemicals,
Chronic exposure to radiation,
Abnormal Immune Mechanisms,
Miscellaneous (PNH, Non-A, Non-B hepatitis)
Drugs Associated with Aplastic Anemia
Anti-bacterial= Chloramphenicol & sulfonamides
Anti-inflammatory = Phenylbutazone, Oxyphenobutazone, Gold salts, Indomethacin
Anti-convulsants = Phenytoin, methoin
diuretics = Chlorothiazide
Anti-thyroid = Propyl/Methylthiouracil, Carbimazole, Thiocyanate, Potassium perchlorate
Oral hypoglycemic = Chlorpropamide
Anti-malarial = Amodaquine, Chloroquine, mepacrine
Diagnostic Criteria for Aplastic Anemia
Bone marrow cellularity
WBC count <500/uL
Plt count <20,000/uL
Anemia with <1% reticulocytes
A blood and bone marrow cancer characterized by pancytopenia with an unknown cause
Acute Lymphoblastic Leukemia (ALL)
Common childhood ALL
L1
Common adulthood ALL
L2
Burkitt type leukemia
L3
An antigen present on the leukemic cells of 70% of ALL patients
CALLA (Common Acute Lymphoblastic Leukemia)
ALL Cytochemical stain rxn
MPO & SBB (-)
ACP (+)
NSE - (+)
TdT- Terminal deoxyribonucleotide transferase(+)
Presence of TdT and HLA-DR only
uALL (uncategorized ALL)
Presence of CALLA, TdT, and HLA-DR
cALL (common ALL)
Has a T cell antigen but lacks sheep erythrocyte receptor (CD2)
Pre-T ALL
Has cytoplasmic immunoglobulin but no surface immunoglobulins (no IgG & IgM)
Pre-B ALL
Presence of T antigens and sheep erythrocyte receptor
T ALL
Presence of B antigens and surface immunoglobulins
B ALL
A rare type of leukemia characterized by splenomegaly and mononuclear cells with hair-like or ruffled projections
Hairy Cell leukemia
Which enzyme is positive in Hairy cell leukemia
Tartrate Resistant Alkaline Phosphatase
Most common malignant disease of plasma cells
Multiple Myeloma
Have the same characteristics as multiple myeloma but is not a malignant condition
Benign Monoclonal Gammopathy
Russell bodies accumulation exhibits what
Grape cell/Mott celllMorula cell
What are the cellular inclusions in Multiple Myeloma
Dutcher Intranuclear bodies
A type of cell that exhibits blue color and mottled appearance
Plasmablast
Crew-cut appearance on skull X-ray is found in
Beta-Thalassemia major
is a structural variant and rare type of δbeta-thalassemia caused by a fusion of δbeta-globin genes
Hemoglobin Lepore
The first type of cell in a developing embryo
RBCs
The most abundant hemoglobin in newborn
HbF
The most abundant hemoglobin in adults
HbA1
first identifiable RBC stage under the light microscope
rubriblast
Last stage of mitosis of RBC presursor
rubricyte
Last nucleated stage in RBC precursor
Metarubricyte
Last stage capable of hemoglobin synthesis in RBC precursor
Reticulocyte
Last erythropoietic stage with a nucleus present
Most common nucleated RBC that interferes with RBC counting
metarubricyte
Last erythropoietic stage where mitosis occurs
Hemoglobin component of RBCs are first detected
rubricyte
Pica is commonly associated with?
IDA
Release by the liver in response to high iron levels
Neutralizes ferroportin
Hepcidin
Aplastic anemia is characterized by ?
Normocytic, normochromic anemia
Other conditions assoc with normocytic, normochromic anemia
Hemolytic anemia
Aplastic anemia
Anemia of chronic blood loss
Myelophthisic anemia
Stem cell related anemia
Abnormal cell seen in uremia
Burr cell
an enzyme that converts phosphoenolpyruvate to pyruvate generating two molecules of adenosine triphosphate; essential for aerobic anaerobic glycolysis
Pyruvate kinase
dark blue granules and filaments in cytoplasm (seen in reticulocytes)
Diffuse basophilia
Dark blue-purple, fine or coarse punctuate granules distributed throughout the cytoplasm
Basophilic stippling
Dark blue-purple dense, round granule; usually one per cell; occasionally multiple
Howell-Jolly bodies
Round, dark blue-purple granule attached to inner RBC membrane
Heinz bodies
Irregular clusters of small, light to dark blue granules, often near periphery of the cell
Pappenheimer bodies
Rings of figure-eights
Cabot rings
Fine, evenly dispersed, dark blue granules; imparts “golf ball” appearance to RBCs
Hb H
Associated conditions with Howell-Jolly bodies
Hyposplenism
Postsplenectomy
Megaloblastic anemia
Hemolytic anemia
Thalassemia
Myelodysplastic syndromes
Associated conditions with Heinz bodies
G6PD deficiency
Unstable Hemoglobins
Oxidant drugs/chemicals
Associated conditions with Basophilic stippling
Lead poisoning
Thalassemias
Hemoglobinopathies
Megaloblastic anemia
Myelodysplastic syndrome
Associated conditions with Pappenheimer bodies
Sideroblastic anemia
Hemoglobinopathies
Thalassemias
Megaloblastic anemia
Myelodysplastic syndromes
Hyposplenism
Postsplenectomy
aka Pentose Phosphate shunt
Hexose Monophosphate Shunt
Pathway that converts ferric iron to reduced ferrous iron
EMB pathway
Nucleus is elongated, curved or sausage-shaped with rounded ends and areas of dense clumping at each pole
band cell/stab cell/ staff cell
Stool containing dark red or black color
Melena
Stool containing fresh blood
Hemotochezia
Blood leakage into the joint cavity
Hemarthrosis
Expectoration of blood
Hemoptysis
Vomiting of blood
Hematemesis
Excessive menstrual bleeding
Mennorhagia
Uterine bleeding not related to menstruation
Metrorrhagia
Platelet Distribution Width (PDW) & Mean Platelet Volume is derived from where
Platelet Histogram
RDW & MCV are derived from where
RBC histogram
The first factor affected by Coumarin therapy
Factor VII
Coagulation factor deficiency that is commonly found in Ashkenazi Jews
Factor XI
What is the effect of microcytosis in PLT count using automated counters?
Falsely elevated PLT count
Most common problem in cell counting causing an increase in current
Positive error
What causes Negative error?
Lysis
Hemoglobin that is insoluble under lowered oxygen tension
Hb S
Oxygen saturation which causes sickling in homozygotes
<85%
Hemoglobin that is resistant to alkali denaturation
Hb F
Conditions associated with increased HbF
Infancy
Hemoglobinopathies
Doubly heterozygous conditions
Thalassemia
Aplastic anemia
Leukemia
Pregnancy
What is the principle of Alkali Denaturation Test
Principle: Hemoglobin is denatured once exposed to an alkali environment
Hb F resists denaturation
Hereditary hypersegmentation of PMNs
Undritz Anomaly
Functional iron
Hemoglobin - blood
Myoglobin - muscles
Peroxidase, catalase, cytochromes, riboflavin enzymes in all cells
Storage form of iron
Ferritin - macrophages
Hemosiderin - hepatocytes
Transport iron
Transferrin
Length of Wintrobe tube
115 mm
Length of Westergren tube
300 mm
Graduation lines of Wintrobe and Westergren
Wintrobe = 100
Westergren = 200
Zetafuge and special capillary tubes are used for?
Zeta Sedimentation Rate
What is M4 called
Naegeli Monocytic Leukemia
What is the FAB and WHO classification of Schilling’s Leukemia
FAB: Cute Monocytic Leukemia
WHO: t9:11
M6
FAB classification: Erythroblastic Leukemia
DiGuglielmo’s Syndrome
Poikilocyte in McLeod phenotype
Acanthocytes
Infectious crisis is the common cause of death in ——
Sickle Cell Disease
A rare disease characterized by impaired oxidative metabolism caused by decreased ability of neutrophils to undergo a respiratory burst after phagocytosis of foreign organisms.
Chronic Granulomatous Disease
What tests are performed to diagnose CGD/
(1) Nitroblue tetrazolium test
(2) Chemiluminescence test
A test that detects Prekallikrein
Activated Partial Thromboplastin time (aPTT)
Hemoglobin that is bound carbon monoxide, which prevents normal oxygen exchange.
carboxyhemoglobin
Irreversible hemoglobin variant
Sulfhemoglobin
Abnormal form of hemoglobin in which the ferrous ion is oxidized to the ferric state; cannot carry oxygen.
Methemoglobin
RBC inclusion: clamshell or bar of gold appearance
Hexagonal crystals with blunt ends that stains deeply
Found within RBC membrane
Hb CC crystal
RBC inclusion: Washington monument appearnace
Hb-SC crystal
What is the FAB Classification of M3
Acute Promyelocytic Leukemia
Most fragile blood cell associated with Chronic Lymphocytic Leukemia
Smudge cells/Basket cell
A taste of primary platelet function
Bleeding time
What is the normal range for platelet
200,000 - 400,000/uL
Platelet aggregometer measures_____?
Change in light transmission
Platelet factor 3 is associated to the activated platelet’s____?
Phospholipid
Stimulates aggregation
Platelet factor 4
Estimated platelets per OIO field
8-20 plts
The outermost zone of platelets
Consists of glycoproteins and surface receptors for coagulation factors
Contains pores for communication between the intracellular and extracellular compartments of PLTs
Glycocalyx
Glycoprotein that binds with vWF
Glycoprotein Ib
Binds with fibrinogen
Glycoprotein IIb-IIIa
Binds with thrombin
Glycoprotein V
Site of prostaglandin and arachidonic acid synthesis
Dense Tubular System
Inhibits platelet activation by increasing cAMP levels
Prostacyclin (PGI2)
Its deficiency results to a normal platelet aggregation with decreased platelet adhesion
12-HETE (hydroxyeicosatetraenoic acid)
An endothelia cell derived chemorepellent which inhibits platelet adhesion
13-HODE (hydroxyoctadecadienoic acid)
Converts plasminogen to plasmin
tPA
Binds and inactivates thrombin
enhances protein C action
Thrombomodulin
Weakly enhances anti-thrombin III activity
Heparan Sulfate
A substance that stimulates vasodilation
Adenosine
A disintegrin and metalloproteinase with thrombospondin type 1 motif, member 13.
Regulates the size of circulating von Willebrand factor by cleaving ultralong VWF multimers into shorter segments that have less hemostatic potential
ADAMTS-13
Recommended for detecting intrinsic and common factor deficiencies and monitoring heparin therapy
aPTT
Normal value of aPTT
20-45 seconds
Used to rule out heparin as the cause of increased thrombin clotting time (TCT). Reverses the effects of heparin.
Protamine Sulfate