Hematology Test 1

Basophil (Baso)

"Blueberry Muffin"

What is the order of migration of hemoglobins ACFS on cellulose acetate, pH 8.6, from the cathode to the anode?

(-) C, S. F, A (+)

Steady state of Bone Marrow can be upset as a result of?

,Genetic mutations ,Poor nutrition ,Medication ,Toxin exposure ,Acute and chronic infections ,Cancers ,Autoimmune conditions ,Immunodeficiency states

Hypo, micro RBCs

Chronic hemorrhage

In the Kleihauer Betke stain, what is the purpose of the citrate buffer at pH 3.3? What is the purpose of the hematoxylin and eosin stains?

Citrate buffer denatures Hb A and leaves Hb F intact The stains allow for visualization of any Hb left in the cells (it is not specific for any one type of Hb)

Segmented Neutrophils - Adult (Males and Females) Reference Range

50-70 Relative % 2.3-8.1 Absolute (x10^9/L)

12. The area of the entire grid on one side of the counting chamber is _______ sq. mm.

9

Name the polypeptide chains found in Hb A, Hb A2 and Hb F?

A = alpha and beta; A2 = alpha and delta; F = alpha and gamma

Normochromic, normocytic RBCs, polychr

Acute hemorrhage

In the Kleihauer Betke stain, what is the appearance of fetal cells vs adult cells?

Adult cells are pale ghosts; fetal cells stain red

Height of voltage pulses

Corresponds to the cell size or volume

Hemosiderin

storage form of iron

What triggers the secretion of EPO?

Hypoxia triggers release from the kidney

Where does hematopoiesis takes place in Fetus?

Takes place in different areas of developing fetus

Hypo, micro RBCs, basostippling, NRBCs

Thalassemia

The following is a Sysmex scatterplot depicting neutrophils, lymphocytes and monocytes.

The X axis =DC The Y axis = RF

Sites for bone marrow collection in Adults

The preferred site for bone marrow aspiration and biopsy in an adult is the Posterior iliac crest. Other sites for adults- sternum and the anterior iliac crest

Urobilinogen

breakdown product of porphyrin ring of hemoglobin

Plasticity

can differentiate into cells they do not normally form (i.e. nerve cells, pancreas cells, muscle cells, etc.)

Anemia of chronic disease

mcv decreased

Aplastic anemia

mcv normal

G-6-P-D deficiency

mcv normal

Meta

means after

Hereditary Spherocytosis

mvc normal

In extravascular hemolysis, indicate if the following are increased, decreased, normal or not present? Serum conjugated bilirubin

normal or slight increase

Eosinophil (Eos)

nucleus looks like 2 lobes, and had red granueles.

Quiescent

undergo cell division only every 3-4 weeks (protection mechanism- the less HSC divide, the less risk of a mutation)

Stem Cell

Capable of cell renewal and differentiation into all required cell lineages

Bone Marrow produces?

-420 billion blood cells per day, or Almost 20 billion cells per hour

Functions of Bone Marrow

-Provides continuous supply of mature blood cells into circulation -Responds to an increase in need for blood cells in abnormal conditions (i.e. infections, anemia) -In response to bacterial infections- bone marrow increases production of neutrophils -In response to anemia- bone marrow increases production of erythrocytes -Removes old and damaged blood cells from the body -Body maintains hemostasis (balance) between cells produced and cells destroyed -Over production or underproduction of cells has clinical consequences that can be life threatening

Basophil-Adult (Males and Females) Reference Range

0-2 R 0-0.1A

Bands- Adults (Males and Females) Reference Range

0-5 Relative % 0-0.6 Absolute (x10^9/L)

12. The area of one small (RBC) square is ______ sq. mm.

0.04

Low POwer WBC

0.2

12. The area of one large (WBC) square is ______ sq. mm.

1

Name three conditions that will cause a false positive retic count?

1) Heinz bodies; 2) Howell-Jolly bodies; 3) Pappenheimer bodies.

Name 4 conditions that will cause a false increase in hemoglobin. Anything that can cause turbidity in the sample will falsely increase the hemoglobin since it is measured by a spectrophotometric method.

1) lipemia (triglycerides > 1000 mg); 2) high WBC count (> 50,000/L); 3) unlyzed RBCs (Hgb crystals in RBCs); 4) abnormal globulins (they precipitate in the reagent causing turbidity).

What is the clinical purpose for doing an ESR?

1) stage and follow rheumatoid arthritis; 2) diagnosis of polymyalgia rheumatica and temporal arteritis.

Function of Lymph Nodes

1. Play role in formation of new lymphocytes 3. Involved in processing of specific immunoglobulins )(Antibodies 4. Filter matter, debris, and bacteria entering the lymph node

Bilirubin

breakdown product of porphyrin ring of hemoglobin

Eosinophil- Adult (Males and Females) Reference Range

1-3 R 0-0.4 A

Types of Bone Marrow Sample

1. Aspiration: Bone marrow is aspirated out of the marrow cavity with a syringe. The liquid sample consists of sinusoidal blood and bone marrow particles 2. Biopsy: A core of bone marrow tissue is removed with a specialized needle

Hepatic Phase

1. Begins at 5-7 gestational weeks and characterized by clusters of developing erythroblasts, granulocytes and monocytes colonizing the fetal liver, thymus, spleen, placenta and ultimately, bone marrow space. 2. Lymphoid cells begin to appear 3. Hematopoiesis during this phase occurs extravascularly 4. Liver remains major site of Hematopoiesis during second trimester of fetal life. 5. Hematopoiesis in fetal liver reaches peak by third month of fetal development, declines after the 6th month, retains minimal activity until 1-2 weeks after birth (graph)

Body's Response to an Increase Demand for Blood Cells.

1. Controlled by growth factors and cytokines 2. When the body needs more cells of a specific type, the body can: Increase the cellularity of the bone marrow, Release developing blood cells earlier from the bone marrow into circulation, Initiate blood cell production in extramedullary sites (i.e. spleen, liver)

Hepatic Phase (Continue)

1. Developing spleen, kidney, thymus, and lymph nodes contribute to Hematopoiesis during this phase 2. Thymus: first fully developed organ in fetus. Major site of T-cell production 3. Spleen and Kidney-major site of B-cell production 4. Production of megakaryocytes 5. Detectable Hemoglobin: Hb F (fetal Hb) is predominant, however, Hb A (adult Hb) may be present

Red Marrow

1. Found in the sternum, skull, scapulae, vertebrae, ribs, pelvic bones and proximal ends of long bones 2. Composed of extravascular cords that contain all developing blood cell lineages, stem and progenitor cells, adventitial cells, and macrophages 3. Mature cells of the bone marrow eventually enter the peripheral circulation by a process not well understood

Mesoblastic Phase

1. Hematopoiesis at the 19th day of embryonic development after fertilization 2. Primitive erythroblasts in yolk sac are important in early embryogenesis to produce Hb (Gower-1,2 and Portland). Need this Hb to deliver oxygen to early developing tissues 3. Yolk Sac Hematopoiesis- intravascularly, within developing blood vessels 4. Precise origin of the adult HSC remains unresolved

Hematopoiesis in Adults

1. In adults, hematopoietic tissue is involved in the proliferation and maturation of blood cells 2. Numerous organs and tissues contribute to this process: Bone marrow, lymph nodes, spleen, liver and thymus 3. Bone marrow contains developing erythroid, myeloid, megakaryocytic and lymphoid cells. 4. Lymphoid development occurs in primary and secondary lymphoid tissue. Primary lymphoid tissue (T, B cells) consists of bone marrow and thymus. Secondary lymphoid tissue consists of the spleen and lymph nodes

Spleen

1. Largest lymphoid organ in the body Vital, but not essential for life. 2. Functions as filter of the circulating blood 3. There are 3 types of splenic tissue: White pulp, red pulp, and marginal zone. 4. White pulp: scattered follicles with germinal centers containing lymphocytes, macrophages and dendritic cells. 5. Marginal zone: surrounds white pulp and contains blood vessels, macrophages and specialized B cells 6. Red pulp: composed of vascular sinusoids and sinuses 7. Splenomegaly occurs often in conditions such as CLL, genetic defects in RBCs, hemoglobinopathies, thalassemia, malaria, etc.

Bone Marrow

1. Largest organ in the body; tissue located within the cavities of the cortical bones 2. There are Two types of marrow: Red Marrow (actively developing cells), and Yellow Marrow (primarily adipocytes) (inactive marrow) 3. During infancy and early childhood, all the bones have red marrow. 4. Between 5 and 7 years of age, adipocytes become more abundant and occupy spaces in long bones. 5. Retrogression- process of replacing active marrow by adipocytes. Results in restriction of active marrow in sternum, vertebrae, scapulae, pelvis, ribs, skull and proximal portion of long bones. 6. Normal adult bone marrow has approximately equal amounts of red and yellow marrow

3 Phases of Hematopoiesis in fetus

1. Mesoblastic Phase (Yolk Sac) 2. Hepatic Phase 3. Medullary (Myeloid) Phase:

Lymph Nodes

1. Organs of lymphatic system 2.Divided into two basic regions: cortex and medulla 3. Further divided into regions: 4. Cortical regions or cortical nodules: B-cell proliferation 5.Paracortex: T-cells and macrophages 6. Medullary cords: lie towards interior of the lymph node and consist of primarily B- lymphocytes and plasma cells

Thymus

1. Originates from endodermal and mesenchymal tissue 2. Populated initially by lymphocytes from yolk sac and liver 4.Very efficient, well-developed organ at birth 4. Size of thymus related to age. Weighs about 12-15 g at birth, increases at puberty to 30-40 g and decreases to 10-15 g at later ages.

Liver

1. Plays significant role in hematopoiesis beginning around second trimester 2. Major site of blood cell production during fetal hepatic stage 3. Major functions in adults: synthesis of various transport proteins, storing of essential minerals and vitamins, conjugates bilirubin from hemoglobin degradation and transports bilirubin for excretion, coagulation factor synthesis, carbohydrate and lipid metabolism 4. Capable of extramedullary hematopoietic production in case of bone marrow shutdown

In the Hemocue method, what is the purpose of the 570nm reading and the 880 nm reading?

1. The product of hemoglobin and the reagent is azidemethemoglobin and it is measured at 570 nm. The reading at 880 nm is subtracted from the 570 nm reading and corrects for turbidity in the sample, similar to a plasma blank.

Medullary Phase

1. Third phase of blood cell production during fetal development 2.Prior to the fifth month of fetal development, hematopoiesis begins in the bone marrow cavity 3. Also known as Medullary Hematopoiesis (occurs in medulla or inner part of bone marrow) 4. Hematopoietic stem cells (HSCs) and mesenchymal cells migrate into the core of the bone marrow and differentiate into structural elements (i.e. stromal cells) 5. Myeloid-to-Erythroid ratio approaches adult levels of 3:1 6. By the end of 24 weeks, bone marrow is primary site of blood cell development

Can unconjugated bilirubin be found in the urine?

1. Unconjugated bilirubin is insoluble in water and does not appear in the urine.

PLT OIl Field

15

Lymphocytes-Newborns (0-1 day old) Reference Range

18-38 R 4.1-8.7 A

Lymphocytes - Adult (Males and Females) Reference Range

18-42 Relative % 0.8-4.8 Absolute (x10^9/L)

Monocytes-Adult (Males and Females) Reference Range

2-11 R 0.45-1.3 A

High Power WBC

2.5

Segmented Neutrophils- Children (1-3 years) Reference Range

22-46 R 1.2-8.9 A

12. The outer dimensions of the entire grid on one side of the counting chamber is ____ mm width x ____ mm length.

3, 3

Segmented Neutrophils - Newborns (0-1 day old) Reference Range

37-67 R 3.7-30.0 A

Lymphocytes - Children (1-3 years) Reference Range

37-73 R 4.2-8.4 A

How many protoporphyrin rings, iron and polypeptide chains are in a hemoglobin molecule?

4 protoporphyrin rings, 4 iron and 4 polypeptide chains

What are the critical values for hemoglobin?

< 7 g/dl or > 20 g/dL (Values beyond these limits severely affect cardiac function)

Flag

An alert generated by an instrument to signal the scientist that a sample has a potential problem that should be investigated

Threshold

An electronic limit that defines which cells will be counted.

Window

An upper and lower threshold

primitive hematopoiesis

Blood cell production occurring during Mesoblastic stage

definitive hematopoiesis

Blood cell production occurring during fetal hepatic stage (continues throughout adult life)

If an instrument is using only impedance technology, how does it differentiate: A red blood cell from a giant platelet

Cannot see

Howell jolly body (composition of inclusion)

DNA (nuclear Fragment)

Iron deficiency anemia

Decreased MCV

Heinz Body (composition of inclusion)

Denatured Hemoglobin

Using appropriate nomenclature, name the stages of the maturation sequence of the erythrocytic, granulocytic, lymphocytic, monocytic, plasmacytic, and megakaryocytic series in order from the undifferentiated cell to the mature cell.

Descrive feom stem cell to mature cell

Schistocytes

Dissem. Intavascular Coag

A lavender tube is kept out on the counter for 6 hours at RT. Which of the following tests will be adversely affected?

Erythrocyte Sedimentation Rate will be falsely decreased

How will Giants Platelets affect platelet count?

False decrease

How will Platelet clums affect platelet count?

False decrease

State 5 conditions that cause false positive and 5 conditions that cause false negative reactions in the dithionite solubility test

False positive: 1) some other Hbs (C-Harlem, C-Georgetown); 2) abnormal proteins that precipitate in the solution; 3) lipemia; 4) polycythemia; 5) recent transfusion with Hb S False negative: 1) anemia with Hb < 7.0 gm/dL; 2) recent transfusion (dilutes out the Hb S; 3) newborns and infants due to decreased synthesis if beta polypeptide chains; 4) other conditions with increased Hb F such as thalassemia; 5) old, deteriorated reagents.

1. Name 5 conditions that would falsely increase the microhematocrit and and 4 conditions that would falsely decrease the microhematocrit?

False: Increase: 1) dehydration; 2) plasma trapping; 3) inadequate centrifugation; 4) allowing the tube to stand > 10 min after centrifugation (cells tend to unpack); 5) including the buffy coat (WBCs) in the measurement; 6) prolonged tourniquet application (causes hemoconcentration); 7) hand-braking the centrifuge (causes the cells to "sling" up the tube). False Decrease: 1) hydration; 2) excess EDTA; 3) improper sealing and loss of blood from the tube (that is why the test is done in duplicate); 4) excessive squeezing of a capillary sample (dilutes the blood with tissue fluid); 5) hemolysis

In order for hemoglobin to be active, the iron must be in what state?

Ferrous, +2

Platelet clumps counted as one large platelet or not counted at all if beyond the upper threshold for counting

Giant platelets - of very large, will exceed the PLT threshold. Severe micro and schisto may be counted as PLTs and falsely increase the count.

relating to the principles of cell differentiation in the Cell-Dyn instrument:Differentiates neutrophils from eosinophils

Granularity (900 degrees scatter) on the y axis vs Lobularity (90 degrees scatter) on the x axis

Heinz bodies

Granulocytes is on the edge

HSC Fates

HSCs are directed to one of three fates: Self-renewal, differentiation, or apoptosis (cell death)

In intravascular hemolysis, what 2 proteins bind to hemoglobin or parts of hemoglobin to prevent its loss through the kidneys?

Haptoglobin and hemopexin

On cellulose acetate at pH 8.6, what other hemoglobins migrate with Hb S, and what other hemoglobins migrate with Hgb C?

Hb D and G migrate the same distance as Hb S. Hb E and O migrate the same distance as Hb C.

Carboxyhemoglobin

Hb with carbon monoxide

What is the normal % of Hemoglobin A and Hemoglobin F at birth?

Hemoglobin A =10-40% Hemoglobin F=50-80%

Hexogonal crystals

Hemoglobin C Disease

Which of the following will be invalid in with in vitro hemolysis of the RBCs in the sample: hemoglobin, hematocrit or both?

Hemoglobin is NOT affected by in vitro hemolysis - remember, the cells have to be hemlyzed anyway in order to measure the hemoglobin. Hematocrit will be falsely decreased with in vitro hemolysis. When the RBCs are lyzed, they make up a less % of the total whole blood volume

Methemoglobin

Hemoglobin with iron in the oxidized or ferric (+3) state

Spherocytes

Hereditary Spherocytosis

Where is bilirubin conjugated with glucuronic acid?

In liver

Where does hematopoiesis takes place in Adults?

In the Bone Marrow

In extravascular hemolysis, indicate if the following are increased, decreased, normal or not present? Serum unconjugated bilirubin

Increased

In extravascular hemolysis, indicate if the following are increased, decreased, normal or not present? Urine urobilinogen

Increased

Pappenheimer bodies (composition of inclusion)

Iron

Hematopoiesis

Is a continuos , regulated process of blood cell production that includes cell renewal, proliferation and maturation. Results in formation, development, and specialization of all functional blood cells that are released from the bone marrow to the circulation

Number of voltage pulses

Is proportional to the number of cells or cell count

Blast

Is the earliest state of a cell lineage

What is the function of erythropoietin?

It stimulates erythropoiesis (increases the rate of maturation and promotes early release of erythroid cells from the bone marrow in response to hypoxia.

relating to the principles of cell differentiation in the Cell-Dyn instrument: Differentiates mononuclear from polynuclear cells

Lobularity 90 degrees scatter on the y axis vs Complexity (7 to 100 degrees scatter) on the x axis

Monocyte (Mono)

Looks like a kidney bean

Thalassemia

MCV Decreases

Pernicious anemia

MCV Increased

Folate or B12 deficiency

MCV Increases

Hemoglobin C disease

MCV is normal

Pro

Means Before

Forward angle scatter ,0 degrees in flow cytometry

Measures cell size

Forward low angle scatter , 2-3 degrees in flow cytometry

Measures cell size

Forward high angle scatter, 5-15 degrees in flow cytometry

Measures internal cell structure

Orthogonal or right angle scatter, 90 degrees in flow cytometry

Measures internal cell structure

In flow cytometry, what is the purpose of hydrodynamic focusing?

Narrows sample stream so that cells enter the counting zone one at a time in single file for counting

What is the name of the stain used for manual reticulocyte counting? What type of stain is it?

New methylene blue; it is a vital stain since it does not have a fixative. It enables visualization of the particulate RNA in the retics.

How will Schistocytes or cell fragments affect platelet count?

No Effect

How will severe microcytosys affect platelet count?

No effect

Hyperseg neut, macroovalocytes

Pernicious Anemia

Hemoglobin C, SC crystals

SC looks like a finger

What is the clinical indication for doing the Sugar Water Test?

Screening for paroxysmal nocturnal hemoglobinuria (PNH)

Polychromatophilia, Polychromasia (Diffuse Basophilia)

Polychromasia/Polychromatophilia - Large red cells (8-10u) which display diffuse basophilia. (blue tinge) due to residual RNA when stained with Wright's stain.

Explain the principle of the cyanmethemoglobin method for hemoglobin

Potasium-ferricyanide xidizes hemoglobin to methemoglobin; K-cyanide converts it to cyanmethemoglobin and is measured at 540 nm.

Hb H Hemoglobin) (composition of inclusion)

Precipitate of beta-globin chains of hemoglobin

basophilia Stipping (composition of inclusion)

Precipitated RNA

In flow cytometry, what is the purpose of the blocker bar?

Prevents non-scattered source light from reaching the detector. Only want to detect scattered light.

How do instruments eliminate interference from red blood cells when counting white blood cells?

RBCs are lysed in the counting fluid.

Diffuse basophilia (composition of inclusion)

RNA

Conductivity or radiofrequency (RF) electromagnetic current resistance

Related to cell size and internal contents

Direct current (DC) resistance

Related to cell size or volume

Cabot Ring (composition of inclusion)

Remnant of mitotic spindle. Looks like a infinity sign inside the cell.

In intravascular hemolysis, what happens to the level of serum haptoglobin and when would you see hemoglobinuria?

Serum haptoglobin levels decrease as it binds to free Hgb and is removed by the liver; hemoglobinuria occurs after the plasma hemoglobin exceeds the renal threshold

How are small particles and debris eliminated from platelet counts?

Setting a low threshold

Sickle cells

Sickle Cell Anemia

What is the defect in Hemoglobin S and C?

Single base substitution at the 6th position of the beta chain of hemoglobin; Hb S has valine instead of glutamic acid; Hb C has lysine instead of glutamic acid

If an instrument is using only impedance technology, how does it differentiate: A red blood cell from a platelet

Size

relating to the principles of cell differentiation in the Cell-Dyn instrument: Differentiates lymphs, monos and basos

Size (0 degrees scatter) on the y axis vs Complexity (7 to 10 degrees scatter) on the x axis

Background

Small particles, debris and electronic noise that is detected in diluting fluids in an instrument when no sample is flowing through the instrument.

How do instruments eliminate interference from white blood cells when counting red blood cells?

They do not, but under normal circumstances, the White Blood Cells are too small in number to have an effect on the RBC count.

In hemoglobin electrophoresis, what causes Hb A and Hb S to migrate differently in the electrical field?

They have different charges

Where and how is urobilinogen formed?

Urobilinogen forms from bilirubin in the intestines through the action of bacteria

Protozoan inclusions

Wen there is an parasitic or bacterial infection

HSC fate process

When a HSC divides, gives rise to two identical daughter cells. Both daughter cells may differentiate (Symmetric division). One daughter Cell may differentiate and the other may return to stem cell pool (Asymmetric division) Or can undergo apoptosis.

In the solubility test for Hemoglobin S, what is the purpose of the following reagents? Sodium dithionite

binds the oxygen in the sample; the reduction in oxygen results in formation of Hgb S crystals causing turbidity of the solution

Sites for bone marrow collection in Children

in a child less than 2 years old is the shaft of the AnteriorTibia. Remember- infants and small children have cellular marrow in all the bone cavities, therefore, the anterior tibia is an appropriate site to sample. All bones in the adults cannot be used for bone marrow collection. For example, the anterior tibia cannot be used for collection, shaft of the tibia will only have fatty (yellow), and not cellular marrow

Active Red Marrow hematopoiesis takes place in?

in the skull, proximal end of large bones, Sternum, Vertebrae, Axial skeleton, Iliac Crest.

Transferrin

iron in transport form bound to apoferritin

If an instrument is using only impedance technology, how does it differentiate: A nucleated red blood cell from a white blood cell:

it cannot

In the solubility test for Hemoglobin S, what is the purpose of the following reagents? Saponin

lyses the RBCs

Ferritin

major storage form of iron