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Red blood cell Information and Courses from MediaLab, Inc.

These are the MediaLab courses that cover Red blood cell and links to relevant pages within the course.

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Laboratories Individuals

Cerebrospinal Fluid
Immature Hematopoietic Cells

The presence of blast cells or immature stages of hematopoietic cell lines are significant abnormal findings in any spinal fluid sample. However, the presence of nucleated red blood cells is the result of bone marrow contamination during the spinal tap.

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Chemical Screening of Urine by Reagent Strip
Hematuria

The term hematuria is used to describe the presence of intact red cells in the urine. The urine may be cloudy/red or pink in color and red cells are visible upon microscopic examination. If the red cells have been destroyed, hemoglobin will be excreted in the urine. The term, hemoglobinuria, is used to describe this condition. The color of the urine will be pink or red but clear rather than cloudy. The presence of only five red blood cells per microliter of urine is considered to be clinically significant. For this reason, a chemical test is needed to detect quantities of blood too small to change the color of the urine. Microscopic examination is used to differentiate between hematuria and hemoglobinuria if the reagent test strip is positive for blood.

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False Negative Results

False negative results may occur with some methods when the concentration of ascorbic acid is greater than 5 mg/dL. The sensitivity of the blood portion of the test strip is decreased in specimens with a high specific gravity and increased protein. High levels of nitrites may delay the reaction, causing a false negative to be reported. If the pH of a urine sample is below 5, hemolysis of red cells as part of the test reaction is inhibited which results in a false negative reaction. An improperly mixed specimen may test negative if the red blood cells are in the sediment.

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False Positive Leukocyte Esterase Test

A false positive result may occur in the presence of strong oxidizing agents in the collection container. In random urine specimens from women, a positive result for leukocyte esterase may be due to a source external to the urinary tract. Other urine sediment findings such as bacteria, squamous or renal epithelial cells, lymphocytes or red blood cells do not contain esterases, and would not produce a positive leukocyte esterase test.

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CLIA Blood Banking Review
IgM antibodies produced against red blood cells generally:View Page
Which of the following red blood cells contain the most H antigen:View Page

CLIA Chemistry / Urinalysis Review
An increased number of these cells, when found upon microscopic examination of urine is termed:View Page
Which of the following is found in plasma but absent in serum:View Page
Increases in LD fractions 4 and 5 are indicative of:View Page

CLIA General Laboratory Review
Match urine color with substance that might have been responsible:View Page

CLIA Hematology / Hemostasis Review
Choose the term that describes the most prominent finding in this peripheral smear:View Page
The red cells in this illustration exhibit which of the following abnormal erythrocyte shapes:View Page
The RBCs found in this illustration are the result of:View Page
The abnormal RBC indicated by the arrow in this illustration is indicative of:View Page
The expected life span of a normal red blood cell is:View Page
Erythropoietin is produced in:View Page
Hemoglobin (g/100ml) x 10 / RBC count (millions/mm3) is the formula for calculating:View Page
An abnormal variation in the size of the red blood cells is termed:View Page
Hypochromia can best be described as:View Page
Which of the following blood smears these illustrations would be best suited for performing a differential count:View Page

CLIA Microbiology / Serology Review
This parasite is found in blood.View Page
I reside in the mouth where I measure approximately 17 micro meters.View Page
I reside inside red blood cells, where I grow and grow until the cells are eventually destroyed.View Page

Erythrocyte Inclusions - Wright Stained Smears
The red blood cells pictured here are normal.View Page

Fundamentals of Hemostasis
Laboratory Tests of Hemostatic Function

Specimen rejection criteria established by your laboratory should be followed at all times, as improperly collected or processed coagulation specimens could adversely affect patient results. Generally speaking, hemolyzed specimens should not be used in coagulation testing because ADP liberated from lysed red blood cells can interfere with a number of coagulation tests, especially those involving platelet assessment. Grossly lipemic specimens may cause erroneous results or a clot may not be detected if a photo-optical coagulation system is used. An alternative method that is not affected by lipemia, such as an electromechanical method, may be required One way to avoid a grossly lipemic specimen is to ask the patient to fast prior to specimen collection.

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Introduction to Bone Marrow
Low Power Magnification

This smear is shown under low power (10x objective) magnification. The reddish cells in the background are mature red blood cells. The dark dots are nucleated erythroid and myeloid precursors. The large dark dot in the middle is a megakaryocyte. Normally, about 5 to 10 megakaryocytes are seen per microscopic field at low power magnification. Clusters of megakaryocytes usually indicate megakaryocytic hyperplasia. Less than 2 megakaryocytes per low power field may mean megakaryocytic hypoplasia.

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Introduction to the ABO Blood Group System
Reverse typing is done using known antisera to detect ABO antigens present on the patient's red blood cells.View Page
The History of the ABO System

In 1900, a German scientist, Karl Landsteiner, discovered that blood groups differ from one individual to another. He took blood samples from five associates and himself, allowed them to clot, and then separated the serum from the cells. Landsteiner found that when he mixed the serum and red cells from different individuals, some samples clumped and some didn’t. Our present day classification of the ABO system is based on Landsteiner’s realization that agglutination occurred because of highly reactive antigens present on the red blood cell which corresponded to antibodies present in the serum. Landsteiner isolated and named the red cell antigens “A” and “B” and the corresponding antibodies “Anti-A” and “Anti-B.” If the red cells contained neither antigen, he called these cells “O”, representing zero antigens present. The fourth type of red cells, “AB”, was discovered in 1902 by Von Decastello and Sturli, associates of Landsteiner. “AB” cells contained both A and B antigens on their surface.

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Rare Subgroups of A

Other rare subgroups of A exist. These comprise less than 1% of the total pool of A genes. Of these rare types, A3 is the most common, but Ax, Aend, and Ael have also been identified. In subgroup A3, the red blood cells characteristically give a “mixed field” agglutination pattern when tested with anti-A and anti-A,B. Small clumps of agglutinated cells are present among large numbers of cells which absorb the antibody to their surface but are not agglutinated by it.

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Agglutination Reactions

Antibodies of the ABO system cause agglutination of saline-suspended red cells at 4°C to 20°C. Heating to 37° weakens the reaction. “Naturally” occurring ABO antibodies may not be strong enough to agglutinate cells without centrifugation. Thus, testing serum for the presence of anti-A or anti-B has classically been performed using the tube system in which serum and cells added to a test tube are centrifuged and then evaluated for agglutination. A slide test has also been performed for forward reactions. Although tube tests are still in wide use, newer systems utilizing other technology such as gel agglutination are becoming more prevalent. The image on this page illustrates agglutination reactions observed with the tube system, from 4+ in the topmost image, to 0 in the lowest image. ABO reactions should be strong. Weak or missing reactions occur, but must be "resolved" before blood products can be released.4+ agglutination: Red blood cell button is a solid agglutinate; clear background.3+ agglutination: Red blood cell button breaks into several large agglutinates; clear background.2+ agglutination: Red blood cell button breaks into many medium-sized agglutinates; clear background; no free red blood cells.1+ agglutination: Red blood cell button breaks into many small clumps barely visible macroscopically; background is turbid; many free red blood cells.Negative: No agglutinated red blood cells present; red cells are observed flowing off the red blood cell button during the process of grading.Other reaction which may occur are the mixed-field reaction, in which mixtures of agglutinated and unagglutinated red blood are present; and hemolysis, in which red cells are hemolyzed by the antibody. Both of these patterns are considered positive reactions.

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Normal Peripheral Blood Cells
Match the characteristics with the cells.View Page
When Lymphocytes Transform

Lymphocytes "transform" in response to antigenic stimuli.Their nuclei becomes larger with more open chromatin and a greater degree of nuclear folding.The cytoplasm becomes abundant, the number of azurophilic granules may be increased and vacuoles may be present.The cytoplasmic membrane may be easily indented by surrounding red blood cells, resulting in a scalloped appearance of the cell's outer edge.These lymphocytes may also be referred to as reactive, activated or stimulated.

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Erythrocytes or Red Blood Cells (RBC's)

The first group is composed of erythrocytes or red blood cells (RBC's). The main function of the erythrocytes is the transport of oxygen from the lungs to the body tissues. Most of the cells in this Wright's stained peripheral blood smear are red cells. On is shown at the arrowhead.

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Glossary of Terms A through M.

Antibody - A modified type of serum globulin synthesized by lymphoid tissue in response to antigenic stimulus. By virtue of specific combining sites each antibody reacts with only one antigen. Anucleate - Having no nucleus. Azurophilic granules - The well-defined large reddish granules (lysosomes) which may be present in large lymphocytes. They are called "azurophilic granules" because they stain blue with the azure stains which were originally used. Basophilic granules - Specific granules present in the cytoplasm of basophils. These granules are large and stain purple-black due to their strong affinity for basic stain. B-cell - Bone marrow derived lymphocytes which produce humoral antibodies. Biconcave - Having two concave surfaces. Cellular Immunity - The capacity of a small proportion of lymphoid population to exhibit response to a specific antigen. Chromomere - The centrally located granular portion of the platelet. Clone - A population of cells descended from a single cell. Delayed Hypersensitivity - (part of cellular immunity) that develops slowly over a period of 24-72 hours after an antigenic stimulus. It consists of an accumulation of cells around small vessels and/or nerves. Example: Tuberculin skin test reaction. Digestive Enzyme - A substance that catalyzes or accelerates the process of digestion. Eosinophilic Granules - Specific granules present in the cytoplasm of eosinophils. These granules are large, refractile spheres which stain reddish-orange due to their strong affinity for acid stain. Erythrocyte (red blood cell, RBC) - One of the elements found in peripheral blood. Normally the mature form is a non-nucleated, circular, biconcave disk adapted to transport respiratory gases. Fixed Macrophage - A phagocyte that is non-motile. Free Macrophage - An ameboid phagocyte present at the site of inflammation. Graft Rejection - A transplanted tissue that is rejected by the body's antibodies. Graft vs. Host Reaction - A complication that occurs when an implanted piece of tissue, which contains antibodies, rejects the host's tissue. Granulocyte - A leukocyte which contains granules in its cytoplasm, i.e., neutrophilic, eosinophilic, or basophilic granules. Half-life - is the length of time it takes for half of the cells circulating at a given time to leave the blood for the tissues. Hemocyte - Any blood cell or formed element of the blood. Hemostasis - A mechanism of the vascular system to arrest an escape of blood. It involves an interaction between blood vessels, platelets, and coagulation. Heparin - A mucopolysaccharide acid which, when present in sufficient amounts, functions as an anticoagulant by inhibiting thrombin. Histamine - A powerful dilator of capillaries and a stimulator of gastric secretions. Humoral Immunity - Acquired immunity produced after response to an antigenic stimulus in which B cells produce circulating antibodies. Hyalomere - the clear, blue non-granular zone surrounding the chromomere of a platelet. Immune Response - The interaction of a cell and an antigen that results in a proliferation of the cell and a capacity to produce antibodies. Isotonic Fluid - A fluid whose elements have an equal osmotic pressure. Leukocyte (white blood cell, WBC) - One of the formed elements of the blood; involved primarily with the body's defense. Lysosome - A microscopic body within cell cytoplasm; contains various enzymes, mainly hydrolytic, which are released upon injury to the cell. Megakaryocyte - A giant cell of the bone marrow from which platelets are derived. Mononuclear - A cell having a single nucleus.

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Identify the nucleated blood cell:View Page
Identify the nucleated blood cell:View Page
Identify the nucleated blood cell:View Page
Identify the nucleated blood cell:View Page
Identify the nucleated blood cell:View Page
Identify the nucleated blood cell:View Page
Identify the nucleated blood cell:View Page
Identify the nucleated blood cell:View Page
Identify the nucleated blood cell:View Page

Phlebotomy
Hemogram (CBC)

Also known as Complete Blood Count (CBC) and is run on whole blood.Blood is tested for quantity and quality of different blood cell types, including: White Blood Cells (WBC Count) Red Blood Cells (RBC Count) Platelets (Platelet Count) Blood is also tested for hemoglobin & hematocrit (H&H).

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Heelstick - specimen collection

Collect the blood into the appropriate tube.Do not: Squeeze the infant’s foot too tightly and wipe with alcohol during the collection.These actions could result in hemolysis (breakdown of the red blood cells), invalidating the test results.

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Hemolysis

Hemolysis means the breakup of fragile red blood cells within the specimen, and the release of their hemoglobin (the red oxygen carrying substance present within the red cells), and other substances, into the plasma.A hemolyzed specimen is one which has undergone hemolysis. A hemolyzed specimen can be recognized after it is centrifuged by the red color of the plasma.

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Plasma

Plasma and formed elements stay mixed in circulating blood. When centrifuged (or spun down), blood is separated into plasma, and formed elements including red blood cells. The plasma separator tube shown here has a barrier to maintain separation of plasma and cellular elements during centrifugation and storage. The red cell layer also includes a relatively small amount of platelets and white blood cells, not visible in the photo on the right.

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Red blood cells

Red blood cells contain hemoglobin, which carries oxygen from the lungs to the tissues of the body. Hemoglobin gives blood its red color. Red blood cells are shown in the photomicrograph of a stained blood smear to the right.

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Reading Gram Stained Direct Smears
Match the following:View Page
In a properly decolorized area, epithelial cells, red blood cells, white blood cells and most background material will stain pink or red. It is in this area that the gram stain reaction of the bacteria can be correctly interpreted.View Page
Red blood cells

Red blood cells are 6-8 microns in diameter and are Gram negative.

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Match the Gram stain reaction for each of the following nonbacterial elements.View Page
In this slide, which structure or structures are present? (Choose ALL of the correct answers)View Page
Overall Procedure

View control smears under oil immersion. If the control smears stained correctly, read the remainder of the smears.Look at the direct smear macroscopically to locate the stained area.Examine the direct smear under oil immersion and find an area that is properly decolorized.Examine at least ten fields in an area that is properly decolorized.Identify the following nonbacterial cell types: epithelial cells, white blood cells, red blood cells, yeast and hyphae.Look for microorganisms and record their characteristics.Quantitate each type of element found and record on the work card.Interpret the direct smear result.Report the direct smear finding.

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Size and Appearance of Cellular Elements

Epithelial cells are larger than white blood cells and red blood cells, and contain a single nucleus. White blood cells (pus cells) usually show a segmented nucleus. Red blood cells are 1/2 to 2/3 as large as white blood cells, contain no nucleus, and are gram negative.Hyphae are gram positive tubular filamentous fungal elements which may show branching or intertwining. Yeast cells are round to oval, often budding, gram positive fungal elements, about the same size as RBCs. They are generally much larger than bacteria.

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Significance of Specific Findings:

Epithelial cells in large numbers within sputum smears means that the specimen is predominantly oral saliva, rather than true sputum from the lung. Epithelial cells in urine smears indicate that the sample has been contaminated by organisms found on the vulva or distal urethra. Bacteria found near or on epithelial cells are usually normal contaminating bacterial flora.White blood cells indicate inflammation and possible infection. The direct smear examination should focus within and around these cells.Red blood cells in a direct smear are not usually significant.Yeast may be present as normal flora in upper respiratory tract or genital tract. They may be significant if they predominate, or if budding yeast forms are seen.Hyphae are more likely to indicate the presence of fungal infection, but this determination requires correlation with clinical findings.Bacteria found in spinal fluid, blood, tissue and specimens from other sterile sites are always significant.Body fluids which are normally sterile must be examined carefully. If only one organism per oil immersion field is identified, then there are about 105 organisms per mL present in the sample! Bacteria observed in specimens from the throat, genital tract and other areas containing normal flora suggest infection only if their composition and type varies significantly from the norm.

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Red Cell Disorders: Peripheral Blood Clues to Nonneoplastic Conditions
The condition most likely associated with the peripheral blood picture shown in the photograph is:View Page
Match the form of red cell inclusions in each of the frames of photographs with a corresponding clinical condition.View Page
Match the red cell shapes in each frame of the photograph with its most likely corresponding clinical condition.View Page
Cells as shown in this iron-stained bone marrow preparation are found in each of the following conditions except:View Page
The nucleated red blood cell and myelocyte photographed here were found on scanning of a peripheral blood smear. In context they are suggestive of metastatic carcinoma to the bone marrow.View Page
The peripheral blood picture is consistent with each of the following conditions except:View Page
G6PD deficiency

A ten-year-old boy came to a physician's attention because of recent jaundice and icteric sclerae. The immediate laboratory work revealed: Hct 24%(normal 36%-47%), MCV 79.5 fl (normal 78-95fl),RDW 13%(normal 11.5-15.0%). His blood smear findings are reflected in these photomicrographs. Note particularly the spherocytes in the upper picture. Some resemble a half-blister with the other half of the cell containing solidly-staining hemoglobin. These are called eccentrocytes. When present, they should trigger a search for red cell hereditary G-6PD deficiency and the oxidant that triggered hemolysis. These morphological findings are only clues; specific testing for G-6PD deficiency should be performed. The blue arrows in the upper photomicrograph are directed toward solid-staining spherocytes in which the cell membrane is beaded by inclusions wrapped within the cell membrane, suggesting the remains of denatured hemoglobin. Included on the smear is a target cell, several acanthocytes, a smudge cell, and a few schistocytes. The lower photomicrograph is supravital staining of affected red blood cells, verifying the presence of Heinz bodies. This disorder was first recognized during the Korean war in 10% of black American soldiers given the antimalarial drug primiquine.

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Reticulocyte identification

Reticulocytes are red blood cells prematurely released from the bone marrow. On a Wright-Giemsa stained blood smear, they appear as polychromatic macrocytes. Their presence in the peripheral blood may suggest hemolysis or bleeding. Their presence is expressed as a percentage of the red cell count: newly born= 3-7%; up to one week of age=1-3%; >one week =0.3-1.8%. Automated or manual methods may be used to enumerate reticulocytes. In clinical context, retics must be separated from debris, precipated stain, Pappenheimer bodies, Howell-Jolly bodies, and Heinz bodies.

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The RBC inclusions shown in the photograph represent which of the following?View Page
Intracellular RBC Inclusions-G6PD (continued)

G6PD deficiency occurs in the same geographic distribution as malaria. It has been theorized that enzyme deficient cells are more resistant to malarial parasites than normal cells.When hemolysis is triggered, the appearance of the red blood cells is modulated by activity of the spleen.Spherocytes, schistocytes, and nucleated red blood cells may appear in the peripheral blood.Denatured hemoglobin removed by an active spleen may leave bite cells, identified by the arrows in this photomicrograph, suggesting the presence of G6PD deficiency.

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Schistocytes vs. bite cells

Schistocyte is a general term for a fragmented red blood cell that may assume various shapes, some with horn-like projections (keratocytes), triangle-forms (triangulocytes), and helmet shapes, as illustrated in the upper photograph. Schistocytes are formed when erythrocytes are forced through a vessel blocked with interlacing fibrin strands and the red cells are sliced into fragments. True schistocytes are devoid of central pallor. These damaged cells continue to circulate while healing their torn edges. Finally, they are removed by the spleen. Bite cells (lower photograph) appear when an abnormal hemoglobin aggregate (Heinz body) is nibbled out of a red cell's cytoplasm by the spleen leaving a bitten apple appearance. Glucose 6-PD deficiency secondary to chemical poisoning or injury by oxidant drugs are settings for Heinz body formation, and the telltale bite cells remain as evidence. Hemolytic anemia associated with severe liver disease is another setting where bite cells are formed.

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The presence of erythrocytes with altered morphology (as photographed here) has a close association with each of the following conditions except:View Page
A 5-year-old girl was brought to a physician's office because of fever and viral-type illness symptoms. Her blood pressure was elevated.Hemogram: hemoglobin 9.1g/dL (normal 12.0 - 16.0 g/dL), hematocrit 28% (normal 37 - 48%), MCV 80 fl (normal 86 - 98 fl), RDW 13.1% (normal 11 - 15%), platelets 90.1 X 109/L (normal 150 - 450 X 109/L) WBC 9.6x109/L (normal 4.3 - 10.8 x 109/L).The peripheral blood smear is represented in the photograph.Which of the following are the most likely associated conditions?View Page
Hemolytic disease of the newborn

Jaundice was recognized in a day-old infant. Notice particularly the size variation (anisocytosis) of the erythrocytes on the infant's peripheral smear. What does this observation mean? Does it provide immediate information that might serve as guidance in expediting diagnosis and treatment? Note that normal-sized red blood cells, microcytes, microspherocytes, macrocytes, and nucleated red blood cells are all present. Red cell variations are expected findings in healthy neonates, but the variations here are exaggerated. Hyposplenic functional features may appear, including acanthocytes, spherocytes, and possibly Howell-Jolly bodies, especially if hemolysis is particularly vigorous. A high (3-7%) reticulocyte count is not unusual during the first three or four days after birth, however, the marrow in this jaundiced infant is proliferating vigorously in response to hemolysis. A call for more red cells is urgent. Immature red cells (in the form of nucleated red cells) and red cells with stippling of RNA (basophilic stippling) are readily identified. Red cell maturation sequence has not been totally processed in the marrow nor is all residual red cell debris removed by the spleen. In the lower photograph are reticulocytes stained by supravital stain (new methylene blue). Basophilic stippling (specks of RNA) stains with both supravital stains and with routine Wright-Giemsa stain.

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Spherocytes and reticulocytes

The photograph represents peripheral blood smear findings in another patient with hereditary spherocytosis. The red cells vary in size (anisocytosis)with a mixture of microcytes (red cells with central pallor) and microspherocytes (red cells with central staining). Macrocytes are conspicuous, some staining light blue. They are immature erythrocytes (reticulocytes)released from the bone marrow early. The bone marrow, geared up for rapid cell release in response to severe hemolysis, expels young red blood cells into the circulation before completing their 24 hour maturation cycle. Hemolysis, jaundice, and gall stone formation disappear following splenectomy. Gallbladder and stone removal eliminate the right upper quadrant pain. A serious consideration, especially in children with hereditary spherocytosis, is hemolytic crisis. A viral infection may allow red blood cell destruction to continue unabated. Anemia of such sudden onset and severity may become catastrophic, with death as the outcome. Splenectomy removes this possibility.

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Warm antibody hemolytic disease

A 49-year-old male with pneumonia was treated with penicillin. He became jaundiced with yellow sclera. Observe the photograph of his peripheral blood smear. Anisocytosis was observed with pale-centered microcytes and polychromatophilic macrocytes. Since penicillin is a classic offender for autoimmune hemolytic disease, the clinician asked for an antihuman globulin (AHG) test, also known as the Coombs test. A positive AHG reaction occurs when the antibody stimulated by penicillin becomes attached to red blood cells. Hemolysis follows, leaving the patient with jaundice and a peripheral blood smear, as demonstrated in the photograph.

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A peripheral smear with red blood cells photographed in a typical field was submitted for review. Which of the following conditions might be eliminated because of the cell population found here?View Page
The photograph is representative of the peripheral blood smear of a five-month-old immigrant from Asia. Her mother was concerned that the child was not eating well. Her spleen was palpable.The hemogram revealed the following:Hb 9.6g/dL (normal 12.0 - 16.0 g/dL)RBC 5.48 X 1012/L (normal 4.2 - 5.9 X 1012/LHCT 30.4% (normal 37 - 48%)MCV 55.4 fl (normal 86 - 98 fl)MCH 17.5 pg (normal 27 - 32 pg)MCHC 31.6 g/dL (normal 31 - 37 g/dL)RDW 34.9% (normal 11 - 15%)Reticulocyte count 10.9% (normal 0.5 - 1.5%)Select the most likely diagnosis based on the clinical information and peripheral blood findings.View Page
Hb E disease (continued)

The family (cited in the previous case history) was from a region of Thailand where the physician knew HbE carriers are prevalent. Homozygous hemoglobin E is common in Southeast Asia and presents with very mild anemia and seldom requires transfusion. Over 30 million people in the world are HbE carriers, making this abnormal hemoglobin almost as common as HbS. Hemoglobin E is uncommon in North America and in Europe, but with changing immigration patterns, hemoglobinopathy E cannot be ignored. Peripheral blood smear findings of target cells, microspherocytes, red cell hypochromia, a few red blood cell fragments, and nucleated red blood cells require evidence from hemoglobin electrophoresis to establish a diagnosis. Clinically, a very important and severe syndrome is hemoglobin E/beta thalassemia in which there is hemolysis requiring repeated transfusions. The patient has a severe anemia, low MCV (50's), and high RBC. This is characteristic of Hgb E/beta thalassemia.

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Dimorphic RBC population

Illustrated in the photomicrograph of a peripheral smear are two populations of erythrocytes. Approximately 50% of the erythrocytes are normal size and contain a full complement of hemoglobin. The patient had received blood transfusions. The transfused red blood cells are the normocytic, normochromic red cells. Admixed are microcytic erythrocytes and larger erythrocytes, some faintly mottled or smudged, suggestive of reticulocytes. This picture represents a hemolytic process with a reticulocyte response. A similar dimorphic red cell population appears following erythropoietin therapy. It is important to recognize when a population of cells in the peripheral smear is not in context with anticipated laboratory findings and the clinical situation.

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A blood smear represented by the photograph was submitted for hematologic review. Based on the erythrocyte morphology and the accompanying histogram, which of the following choices is the most likely situation or condition?View Page
A frail 85-year-old woman living in an extended care facility was found lying on the floor. Her eating habits had been irregular and food intake scanty. Her skin had tissue paper-like quality, with a pearly grey sheen. In good light a faint lemon-yellow color became evident.Her hemoglobin was 9.2 mg/dl. The peripheral blood smear (upper and lower photographs) is most consistent with:View Page
The blood study from which this smear was obtained revealed an MCV of 115 femtoliters (fl).Normal MCV values in adults= 80 - 90 fl.Normal MCV values in full-term infants= 98 -108 fl.Which of the following conditions may be indicated by the results seen on this peripheral blood smear?View Page
References

Glassy, Eric F.,(Ed). Color Atlas of Hematology: An Illustrated Field Guide Based on Proficiency Testing. 1998. College of American Pathologists Hematology and Cliical Microbiology Research Committee. College of American Pathologists, Northfield, IL 60093-2750.Hookey,L., Dexter, D., Lee,D. H. The Use and Interpretation Of Quantative Terminology In Reporting Red Blood Cell Morphology. Laboratory Hematology 7:85-88, 2001.Peterson P, Blomberg DJ, Rabinovitch A, Cornbleet PJ. Physician Review of the Peripheral Blood Smear: When and Why. For the Hematology and Clinical Microscopy Resource Committee of the College of American Pathologists. Laboratory Hematology 7:175-179, 2001

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Guidelines for standard reports

In a study on the reporting of red blood cell morphology abnormalities conducted in Ontario, Canada (Hookey L, Dexter D, Lee DH, Laboratory Hematology 7:83-88, 2001), fewer than 50% of 33 participants used the same term to describe the quantitative frequency of peripheral blood abnormalities. Seven blood smears, each containing one of several abnormal erythrocytes-- schistocytes, teardrop cells, acanthocytes, and Howell-Jolly bodies--were evaluated by 32 participants. The participants were asked to document their evaluations from a list of quantitative terms. There was a heterogeneity in the use of terms "rare," "slight," "occasional," "few," "mild", "present," "moderate," "many," and "marked." Choices of terms were subjective without points of reference. Guidelines for establishing standardized qualitative estimations of abnormal erythrocytes in the peripheral smear are presented as follows: 1+ = 2 - 4/Oil Immersion Field (OIF) 2+ = 5 - 7/OIF 3+ = 8 - 10/OIF 4+ = >10/OIF. The terms "few," "moderate," "many," and "marked" may be substituted for the 1+ - 4+ grading system, but only when their specific points of reference are universally understood in tandem with the above guidelines. A comment should be triggered if any erythrocyte abnormalities are seen in numbers >3/OIF including, but not limited to, polychromasia, basophilic stippling, nucleated RBC's, and Howell-Jolly bodies. Rouleaux or RBC agglutination are important findings and must be documented.

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Normoblasts

Many of the distorted erythrocytes displayed on the previous page are also present on this one. We see anisocytosis, poikilocytosis, fragmented forms, target cells, and a few Howell-Jolly bodies. Note also circulating nucleated red blood cells (normoblasts). The presence of these normoblasts may represent a premature release from a hyperplastic marrow or, more likely, are due to a lessening of the normal inhibition of erythroid release from the marrow as a result of splenectomy, permitting their earlier entry into the circulation.

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Red Cell Morphology
Normal Red Cells

Normal red cells are seen in this field. Mature erythrocytes can be described as round, elastic, non-nucleated, bi-concave discs which appear buff colored on Wright's stained smears. Notice that many of the cells have an area of central pallor which covers about one-third of the cell. The pallor occurs as a result of the disc-shaped cells being spread on the slide. Normal mature red blood cells have an average diameter of 7.2 microns with a range of 6-9 microns. This is approximately the same size as the nucleus of a small lymphocyte, which is often used as a guideline when determining the size of the red cells on a slide. The average thickness of a normal mature red blood cell is 2.1 microns with a mean corpuscular volume (MCV) of 87 cubic mircons/femtoliters.

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Summary of Anisocytosis

Anisocytosis is a general term reflecting increased variation in the size of red blood cells. The MCV will be within normal limits, but RDW will be increased. Variation usually affects a continuum of red cell sizes, but occasionally two distinct red cell populations can be observed(for example in sideroblastic anemia, or after red cell transfusion.)

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Size Variation

Red blood cells can vary in size (diameter/volume) from smaller than normal, microcytes, to larger than normal, macrocytes. When red cells of normal size, microcytes and macrocytes are present in the same field, the term anisocytosis is used.Since the purpose of this unit is to acquaint you with the appearance (identification) of abnormal red cell morphology, percentages of abnormalities present will not be considered. It is important to be aware that rating red cell morphology for the purpose of reporting it is a skill which must be learned before you are able to complete this aspect of a differential count.

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Erythrocytes

Erythrocytes, when spread on a glass slide, show varying degrees of central pallor as noted in the previous exercise. This central pallor is related to the hemoglobin concentration present in the red cells.When viewing normal mature red cells, the central area (one-third of the cell) is white, while buff-colored hemoglobin is visible in the outer two-thirds of the cell. The mean corpuscular hemoglobin concentration (MCHC, 32-36 gm/dl of red blood cells), is the indice value which is used to verify the presence of adequate hemoglobin concentration in the cells visible on the peripheral smear.

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Semen Analysis
Appearance of semen

The color of semen may vary. Normal values are white, yellowish, or gray. Normally the consistency ranges from clear to cloudy. A pink or reddish discoloration suggests the presence of red blood cells, an abnormal finding.

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The Urine Microscopic: Microscopic Analysis of Urine Sediment
True or false? Red blood cells are present in this slide.View Page
Red Cell Casts

Red cell casts appear as clear cylinders containing red blood cells and may have an orange red tinge. Their presence indicates bleeding into the nephron. Red cells within the cast are rapidly hemolyzed and the cast becomes a hemoglobin cast, having an orange color and a homogeneous ground-glass texture. In order for a cast to be considered an RBC cast, the outline of the red cells must be clearly visible in at least one area of the cast.

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Microscopic Examination of Urine Sediment

The sediment may be examined using both brightfield and phase-contrast microscopy. With the brightfield microscope, subdued light must be used. Some structures will be missed if there is too much light in the field. Fine focus throughout the examination to identify structures in different focal planes. Scan the slide on low power for quantification of casts, crystals and elements that are present in only a few fields. Use high power to identify casts and count red blood cells, white blood cells and epithelial cells.

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Cells Types Observed in Urine Sediment

Cells which may be present in the urine include epithelial cells, white blood cells (WBC) and red blood cells (RBC). The epithelial cells in the urine may originate from any site in the genitourinary tract. It is normal to find a few epithelial cells in the sediment. White blood cells may enter the urinary tract anywhere from the glomerulus to the urethra. The WBCs are mostly neutrophils. Red blood cells may originate in any part of the urinary tract. Normally, RBCs do not appear in the urine, although the presence of a few RBCs is not considered abnormal.

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Red Blood Cells

Red blood cells (RBCs) may also be found in the urine sediment. The presence of RBCs in the sediment is associated with damage to the glomerular membrane or vascular injury within the genitourinary tract (the possibility of menstrual contamination must be considered).

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Red Blood Cells versus Yeast

In this field both red blood cells and yeast are present. Since red blood cells are readily hemolyzed by dilute acetic acid, a drop is allowed to flow under the edge of the coverslip. The acetic acid lyses the red blood cells, leaving only the remaining yeast.

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Lysis of Red Blood Cells with Acetic Acid

This slide shows residual yeast after red blood cells have been lysed with acetic acid.

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A 10% solution of acetic acid can be used to differentiate what from red blood cells?View Page
A technologist is trying to enumerate the number of fat droplets in a urine sediment which also contains numerous red blood cells. Which of the following reagents would stain the fat droplets orange red?View Page
Fat Droplets

Fat droplets can be mistaken for red blood cells. Sudan III will selectively stain the neutral fats bright red orange. Cholesterol can be identified by Maltese cross formation under polarized light.

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White Cell and Platelet Disorders: Peripheral Blood Clues to Nonneoplastic Conditions
Match the letter representing the cell type with the condition in which increased numbers of the cell may be found in the peripheral smear.View Page
Peripheral blood smear preparation

A reproducible blood smear review requires every peripheral smear be prepared for consistent openness and clarity. Consistency is maintained by uniform handling of every blood smear. Good results may be expected when the preparation is begun with only a small drop of blood at one end of a clean glass slide. The drop is smeared lightly and quickly so as to leave a thin (feathery) edge where all cells may be examined individually, particularly red blood cells. The site of examination then is chosen away from clumping, piling, or bumping of cells against each other, perhaps a site five or six oil fields from the end of the feathery portion. Such an area for examination is illustrated in the photograph.

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The cell bulging with inclusions in the image on the right is most consistent with Chediak-Higashi anomaly.View Page


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