Hematology Exam 1 Flashcards
Lecture 1-4
What is hematology?
Study of blood cell production, destruction, number, and disease.
Normal WBC value
3.6-10.6
Normal RBC value for men
4.20-6.00
Normal RBC value for women
3.80-5.20
What is the difference between serum and plasma?
Plasma doesn’t clot due to anticoagulant stopping the coagulation cascade. Serum doesn’t contain clotting factors.
Primary specimen source for hematology
Whole blood
Most common anticoagulants used in hematology?
EDTA and Sodium Citrate (rarely heparin is used but it does not stain well so you can’t use it for smears)
What types of cells will you find in peripheral blood?
Erythrocytes (RBCs), thrombocytes (Platelets/PLTs), leukocytes (WBCs: <=5 banded neutrophils, segmented neutrophils, lymphocytes, monocytes, eosinophils, basophils)
Lifespan of RBCs
Up to 120 days
“cyte”
Cell
Intra-
Within
Medullary
Bone marrow
-lysis
Break-down
Extra-
Outside
Intramedullary hemolysis
Cell break-down within bone marrow
Extramedullary hemolysis
Cell break-down outside bone marrow
Erythroid hyperplasia
Increased red cell formation
Lymphopenia
Decreased number of lymphocytes
Eosinophila
Increased number of Eosinophils
Polycythemia
Many cells increased (all cells in the blood)
Vascular
Vessel
-penia
Decreased
Cytosis, -philia
Increased cell number
This is the term that indicates RBC size
MCV - Mean Cell Volume
This is the term for the weight of hemoglobin contained in RBCs
MCH - Mean Cell Hemoglobin
Term for average concentration of hemoglobin in RBC
MCHC - Mean Cell Hemoglobin Concentration
What occurs if both MCH and MCHC are decreased?
Microcytic anemia
What occurs if both MCH and MCHC are increased?
Macrocytic anemia
What is included in a CBC?
WBC, RBC, Platelet count, Hemoglobin, Hematocrit, and RBC indices
What is the Reticuloendothelial System (RES)?
Network of cells and tissues in spleen, liver, thymus, bone marrow, and lymph nodes
The purpose of the RES
Network involved in the formation and destruction of blood cells, metabolism of iron, inflammation, and immunity
This network provides cells to fight infection, carry oxygen, and prevent bleeding
Reticuloendothelial System
One of the largest organs in the body
Bone marrow
Tissue located within the cavities of the cortical bones which consist of trabecular bone resembling a honeycomb-like structure
Bone marrow
Two types of bones marrow
Red and Yellow
Ratio of red marrow to yellow marrow in adults
50:50
Hematopoietically active marrow that produces cells
Red marrow
Hematopoietically inactive marrow composed primarily of adipocytes (fat cells)
Yellow marrow
True or False:
Sites of hematopoiesis increase during fetal development
False- The sites decrease
The age that retrogression occurs for bone marrow locations
Between 5-7
Where red marrow is restricted to after retrogression occurs
Flat bones: sternum, pelvis, ribs, and skull.
Vertebrae
Proximal ends of the humerus and femur
What Red Marrow is composed of
Extravascular cords that contain all of the developing blood cell lineages, stem and progenitor cells, adventitial cells, and macrophages
Where RBCs develop in Bone Marrow
Small clusters adjacent to the outer surfaces of the vascular sinuses
Where megakaryocytes develop in Bone Marrow
Close to the vascular walls
RBC precursors surround this cell type that carries iron to incorporate iron into hemoglobin
Macrophages
Cells that contract allowing mature blood cells to pass through pores of the endothelial cytoplasm and into circulation
Adventitial Cells
Organ that produces erythropoietin
Kidneys
Bean shaped structures that occur in groups or chains in various intervals that run parallel to but NOT part of the circulatory system
Lymph nodes
1st main function of lymph nodes
Play a role in the formation of new lymphocytes
2nd main function of lymph nodes
Involved in the processing of specific immunoglobulins (antibodies)
3rd main function of lymph nodes
Involved in the filtration of particulate matter, debris, and bacteria (causing the inflammation we feel with things like strep throat)
Largest lymphoid organ in the body
Spleen
Organ that technically is not essential for life
Spleen
Function of the spleen
indiscriminate filter of the circulating blood
The spleen filters this much blood per minute
350mL/min
The spleen contains about how much blood?
350mL
Sponge-like filter composed of an inner “white” pulp and outer “red” pulp
Spleen
White pulp of spleen contains what cells?
Lymphocytes, macrophages, and dendritic cells
This zone surrounds the white pulp of the spleen
Middle Marginal Zone
The middle marginal zone contains these cells
Macrophages and memory T & B cells
The outer red pulp of the spleen is composed of what specialized cell?
Macrophages
What must RBCs be able to do in order to re-enter circulation from the spleen?
Deform
The spleen is the holding area for what percentage of the platelet population in the peripheral blood?
30%
It takes this many days for RBCs to proliferate and mature
5-7 days
RBCs lifespan
120 days
Amount of time for WBCs to proliferate and mature
12-20 days
Number of days for platelets to proliferate and mature
8-9 days
Cell type that should never be seen outside of bone marrow
Megakaryocyte
The cell that cell production starts with
Pluripotential stem cell
A pluripotential cell will proliferate into this cell
Multipotential
Cell capable of self-renewal and has the ability to reconstitute the hematopoietic system of a lethally irradiated host
Pluripotential Stem Cell
CFU
Colony-forming unit
CFU-S
Colony-forming unit - spleen
CFU-S =
CFU-GEMM
CSF
Colony-Stimulating Factor
Cell that commits to only one cell line
Multipotential/Progenitor cell
Multipotential/progenitor cells depend on these growth factors to decide which cell line they become (4 types)
- Erythropoietin
- CSF (colony-stimulating factor)
- Interleukins
- Cytokines
These cells will continue into the tissue
Macrophage and Mast cells
Blood oxygen tension causes the kidneys to produce what growth factor?
Erythropoietin
Primary granules
Granules that:
- Appear 1st
- Appear in promyelocyte
- Are purple-red
- Decrease as the cell matures
Secondary granules
Granules that:
- Being to occur in the myelocyte
- Increase as the cell matures
Eosinophilic granules will stain what color and why?
Red (acidic)
Neutrophilic granules will stain what color and why?
Purple (acid and alkaline)
Basophilic granules will stain what color and why?
Blue (alkaline)
Main purpose of a mature neutrophil
Phagocytosis of bacteria
Name this cell:
- observed only in the bone marrow (may be observed in peripheral blood with leukemia)
- nucleus is round or oval with abundant, unclumped, light purple reticulated chromatin
- 2-5 nucleoli
- moderate basophilic cytoplasm (blue-gray)
- N:C ratio 4:1
Myeloblast
Name this cell:
- observed in bone marrow (may be observed in peripheral blood due to leukemia or severe infection)
- about the size of blasts
- nucleus is round or oval, slight chromatin clumping
- 1-2 nucleoli - beginning to fade
- cytoplasm more evident
- primary granules are observed
Promyelocyte
Name this cell:
- observed in bone marrow (may be observed in peripheral blood due to leukemia or severe infection)
- round or oval nucleus with chromatin getting coarser and may be slightly flattened on one side
- possible “dawn of neutrophilia” seen (clearing near nucleus)
- NO nuclei present
- cytoplasm contains primary and secondary granules and is becoming less blue
- LAST CELL TO DIVIDE
Myelocyte
First stage of cell to differentiate into eosinophil, basophil, or neutrophil
Myelocyte
Name this cell:
- Observed in bone marrow or peripheral blood due to infections
- smaller than myelocyte
- kidney-bean shaped nucleus with chromatin becoming coarser and staining darker
- secondary granules are more evident with cytoplasm being more pink
Metamyelocyte
Name this cell:
- smaller
- first cell normally seen in peripheral blood
- nucleus is horse-shoe shaped and very clumped
- <5% in peripheral blood
- abundant cytoplasm with many fine granules, more secondary granules are evident
Neutrophilic band
Name this cell:
- purpose is to engulf and destroy bacteria
- 50-70% circulate in peripheral blood
- Capable of diapedesis and phagocytosis
- nucleus is pinched into segments connected with a fine filament creating 3-5 loves
- abundant pink cytoplasm with secondary granules
Segmented neutrophil
Name this cell:
- main purpose is humoral and cellular immunity
- nucleus is round, oval
- highest N:C ratio other than a blast
- Chromatin is evenly coarse with dark clumping
- Nucleolus may be seen
- cytoplasm is blue and may have azurophilic granules
- 20-40% in peripheral blood
Lymphocyte
Precursors for lymphocytes
Lymphoblast and prolymphocyte
Differentiates in to T and B cells
Lymphocytes
Name this cell:
- Main purpose is processing antigens to present to lymphocytes and collect garbage (scavengers/”clean-up crew”)
- Become macrophages when they leave peripheral blood and enter tissues
- Largest cell in normal blood
- Nucleus is indented, folded brain-like convolutions, pale chromatin with abundant and distant open spaces, “lace-like”, usually no nuclei
- Gray or gray-blue cytoplasm with pseudopods
- May contain vacuoles
- <10% in peripheral blood
Monocyte
Precursors for monocytes
Monoblasts and promonocytes
Name this cell:
- Purpose: immunity against parasites, control of inflammation, and allergic responses
- Large orange-red cytoplasmic granules due to acidity
- Banded or bi-lobed nucleus
- <5% found in peripheral blood
Eosinophil
Name this cell:
- Involved in cell mediated immunity (interacts with IgE)
- huge dark blue or black granules that contain histamine and are so large they may hide the nucleus
- <2% seen in peripheral blood
Basophil
Name this cell:
-Main function is to produce platelets for clotting blood
Megakaryocyte
Precursors of megakaryocyte
Megakaryoblast, promegakaryocyte
Name this cell:
- Main purpose is to make antibodies
- nucleus is round and eccentric with clumped chromatin
- cytoplasm is dark blue with pale perinuclear zone
- comes from B cell
Plasma cell
Name this cell:
- Purpose: immunity, phagocytosis, storage of iron
- only seen in bone marrow and tissues
- small nucleus in large amount of cytoplasm
- often observe vacuoles or particle matter
- named based on what tissue type they enter
Macrophage
Destroys bone
Osteoclast
Makes bone
Osteoblast
Time in which a peripheral blood smear should be made
No more than 5-6 hours, preferably 3-4
Wright-Giemsa stain:
-This stains acidic cellular structures blue
Alkaline methylene blue
Wright-Giemsa stain:
-This stains basic hemoglobin and eosinophilic granules red
Acidic eosin
Your slide came out too blue with the Wright’s stain, what’s your solution?
- decrease staining time
- decrease pH
- increase buffer time/amount
Your slide came out too pink with the Wright’s stain, what’s your solution?
- increase staining time
- increase pH
- decrease buffer time
Your slide has water artifacts, what’s your solution?
- replace methanol
- keep dishes covered to prevent water absorption
- check humidity
- increase drying speed
If sodium citrate tube (blue) is used for a CBC what must you multiply by to get a correct platelet and WBC count?
1.1
More immature WBCs seen in a differential is referred to as
Left shift
Erythrocytosis or polycythemia (are due to)
Heart/lung problems, dehydration, or myeloproliferative disorder (cancer)
Thrombocytosis is caused by
Infections, inflammation, or rare malignancy
Leukopenia is due to
Severe infections, blood malignancies, certain types of anemia, chemotherapy
Anemia is due to
Blood loss, nutritional deficiencies, inherited conditions
Thrombocytopenia is caused by
Blood malignancies, chemotherapy, and coagulation disorders.
Normal platelet reference range
150,000-450,000/uL
Normal male RBC range
4.2-6.0 x 10^6/uL
Normal female RBC range
3.8-5.2 x 10^6/uL
Normal newborn RBC range
4.1-6.1 x 10^6/uL
What causes males to have a higher RBC value?
Androgen
Total number of individual WBC in 1 uL of blood
-relative percentage x total WBC count = # cells in 1 uL of blood
Absolute Count
Percentage based on a total of 100 WBC
Relative Count
Depth of hemocytometer
0.1 mm
Number of “large” squares counted for WBC count
4 or 9 (low cell counts)
Where are RBCs and PLTs counted on the hemocytometer?
Middle square
How many small squares are in the middle square?
25
What squares are platelets counted on the hemocytometer?
All 25 of the middle square
What squares are RBCs counted on the hemocytometer?
4 corners and very center of the middle square
Measurement of single small middle square
0.2 mm
Measurement of single small square in WBC count of hemocytometer
0.25 mm
Measurement of one large square used to count WBCs in hemocytometer
1 mm
Typical dilution ratio for WBC manual cell count
1:20 (4 squares) or 1:100 (9 squares)
Dilution ratio used for RBC and PLT manual counts
1:100
Usual Diluting fluid used for WBC and PLT estimates
1% ammonium oxalate
Manual counts: DO NOT count which sides of each square?
Right and Bottom
Manual counts:
Must agree within what %?
10%
Calculation for manual count
Total count = cells counted x dilution factor (20 or 100) x depth (10) / area counted based on 1 large square
What may cause an error in manual counting?
- Inadequate mixing or dilution
- non-random distribution
- observer bias
- Incorrect calculation
This principle uses cell impedance
Coulter Principle
When do you have to perform a correction calculation for WBCs?
When NRBCs are >5
Calculation for corrected WBC
WBC x 100 / #NRBC + 100
Primary responsibility of RBCs
- Carry oxygen from lung to tissues
- Return CO2 to lungs
Where does erythropoiesis occur?
Bone marrow
Earliest recognizable erythrocyte precursors in bone marrow
Pronormoblasts
Pronormoblast is also known as
Rubriblast
Name the cell:
- 1% of bone marrow cells
- Earliest recognizable cell of erythroid series
- homogeneous nucleus with nucleoli and chromatin strands
- nucleus stains reddish-blue
- royal blue/navy cytoplasm
Pronormoblast/Rubriblast
Basophilic normoblast is also known as
Prorubricyte
Name the cell:
- nuclear material has begun to coarsen (nucleoli not evident)
- parachromatin evident (more RNA and hemoglobin)
- cytoplasm may have patches of pink (may see perinuclear halo)
- 1-4% of bone marrow cells
Basophilic normoblast/Prorubricyte
Polychromatic normoblast is also known as
Rubricyte
Name the cell:
- nuclear chromatin is thickened and irregularly condensed with pyknotic areas
- no nucleoli visible
- eccentric nucleus
- gray-blue cytoplasm
- 10-20% bone marrow cells
- may be seen in newborns
Polychromatic normoblast/ Rubricyte
Orthochromatic normoblast is also known as
Metarubricyte
Name the cell:
- Incapable of further DNA synthesis
- nucleus is small and dense, round or bizarre shaped, will soon be extruded
- Cytoplasm is blue-pink-orange (increased with hemoglobin synthesis)
- 5-10% bone marrow cells
- Not normally in peripheral blood of adults
- small number may be seen in newborns
Orthochromatic normoblast/Metarubricyte
Polychromatic erythrocyte is also known as
Reticulocyte
Name the cell:
- Nucleus has been extruded
- cytoplasm is diffusely basophilic, polychromatophilic (due to RNA causing blue color)
- larger than mature erythrocyte
- Contains 2/3 of total hemoglobin
Polychromatic erythrocyte/Reticulocyte
Polychromatic erythrocyte is called what when stained with Wright’s stain
Polychromasia
Polychromatic erythrocyte is called what when stained with New Methylene Blue
Reticulocyte
Name the cell:
- Thin central pillow
- biconcave disc
- pliable, deformable
- pink to orange color
- does not synthesize protein
- transports oxygen to tissue
Erythrocyte (Mature RBC)
What % of cell hemoglobin is made during the normoblast stages?
65%
65% of cell hemoglobin is made during what stage of the erythroid proliferation?
Normoblast
What % of cell hemoglobin is made during the reticulocyte (polychromatic) stage?
35%
35% of cell hemoglobin is made during what stage of the erythroid proliferation?
Reticulocyte (polychromatic)
What % of aged erythrocyte destruction is extravascular?
90%
Where does about 90% of aged erythrocyte destruction happen?
Extravascularly
What % of aged erythrocyte destruction is intravascular?
10%
Where does about 10% of aged erythrocyte destruction happen?
Intravascularly
What is returned by transferrin to bone marrow?
Iron
What is transferrin?
Carrier protein
What is protoporphyrin broken down to and where?
Bilirubin in the liver
Approximately how much iron and RBCs are removed from circulation on a daily basis?
1%
