Hematology: Basic Overview Online Training

Continue Continue Untitled Scene Master Template HOOD05162003052540 | Effective Date: 26-Nov-2019 ? Hematology Basic Overview Online Training Hematology is the science that studies blood morphology and blood forming tissues. The quantity and characteristics of blood cells can change depending on the disease condition or treatment. Hematology testing plays an important role in patient care. Identify the different blood cells and their functions Identify analytical methods used in the hematology lab Recognize common tests performed in hematology Recognize procedures for the proper handling of hematology specimens 1 4 3 2 Welcome hematology_welcome.mp3 Welcome to the Hematology Basic Overview Online Training course. Blood is composed of both liquid and cellular elements. While chemistry and immunoassay deal with the chemical components found in the liquid portion of the blood, hematology is concerned with the analysis of the cellular elements. Since the number and characteristics of blood cells can change depending on disease condition or treatment, hematology testing plays an important role in patient care. Knowledge of the tests performed and procedures used in hematology will give you an appreciation of the need for accurate and timely sample analysis in this section of the clinical lab. This course will cover the following four learning objectives. For navigation help, select the question mark button located in the lower right corner of each slide. ? What is Hematology? Hematology is the study of the number, characteristics and functions of the cells of the blood and blood forming tissue. The three types of cells found in the peripheral blood are: White Blood Cells or WBCs Red Blood Cells or RBCs Platelets Bone Marrow is the main blood cell forming tissue. Select the underlined words above to identify the objects in the image. What is Hematology? hematology_whatisheme.mp3 Hematology is the study of the number, characteristics and functions of the cells of the blood and blood forming tissue. There are three types of cells found in the peripheral blood: White Blood Cells or WBCs, Red Blood Cells or RBCs and Platelets. All three of these cells have their beginnings in a tissue located inside the bones called the bone marrow. Select the underlined words to identify the objects in the image. ? Parts of a Cell Cells are composed of numerous components. The main components covered in this course are: Membrane – composed of a complex lipid and protein structure that holds the contents of the cell together Cytoplasm – the material within the body of the cell that is outside the nucleus. It contains different organelles responsible for cellular metabolism and functioning Granules – packets within the cytoplasm that contain enzymes, proteins and other chemicals used for cell functioning Nucleus – contains the cell's (genetic material (DNA and RNA) and controls its growth, metabolism, and reproduction 1 2 3 4 Parts of a Cell hematology_partscell.mp3 Cells are composed of numerous components. The four main components include the membrane, cytoplasm, granules, and nucleus. ? Bone Marrow Bone Marrow is located inside specific bones of the body. It responds to the body's needs to produce different blood cell types Cells mature inside the bone marrow from very young stem cells (blasts) into mature forms that are released into the peripheral blood Cells may be released prematurely into peripheral blood in disease conditions such as anemia and leukemia Bone Marrow hematology_bonemarrow.mp3 The bone marrow is located inside specific bones of the body and responds to the body needs by producing different types of blood cells. Cells develop over time from very young cells, called blasts, into their mature forms within the bone marrow. Once cells are mature, they are released from the bone marrow into the blood. In some disease conditions such as severe infections, anemia and leukemia, immature cells can be released prematurely from the marrow and can be found circulating in the peripheral blood. ? White Blood Cells White Blood Cells (WBCs) are: A major component of the immune system which defends the body against invasion by foreign organisms or chemicals Produced during infections or other conditions requiring an immune response Select the tab arrows to learn more about the different types of WBCs. Neutrophils Lymphocytes Monocytes Eosinophils Basophils Which cells are WBCs again? White Blood Cells hematology_wbc.mp3 White blood cells are a major component of the immune system which defends the body against invasion by foreign organisms or chemicals. They are produced during infections or other conditions requiring an immune response. There are five different types of WBCs normally found in blood. Select the tab arrows to learn more about the different types of WBCs. Basophils Basophils (Basos), similar to Eosinophils, contain histamine responsible for many allergic symptoms. They are present in blood in very small numbers but increase in cases of leukemia, chronic inflammation, hypersensitivity reaction to food, or radiation therapy. Eosinophils Eosinophils (Eos) typically are present in the blood in small numbers. They increase in the blood during allergic reactions, skin inflammation and infection with parasites. Monocytes Monocytes (Monos) travel in the blood and the body tissues to engulf dead WBCs and antigens after an immune response. They increase in infection, inflammation, and certain types of leukemia. Lymphocytes Lymphocytes (Lymphs) initially detect the invading substance, or antigen, and alert the other immune cells. Lymphs also: Produce proteins known as antibodies that bind the antigen to inactivate it Are the second most common WBC in the blood, but also are found in the lymph vessels and lymph nodes Increase in number during a viral infection   Neutrophils Neutrophils (Neuts) are the most common WBC seen in blood. Their main role is to travel throughout the blood and tissues to engulf and destroy invading organisms. They increase in number with bacterial infections or tissue inflammation. ? Red Blood Cells Red Blood Cells (RBCs) transport oxygen to tissue cells. To accomplish this, they: Contain Hemoglobin (Hgb) which carries the oxygen Are normally biconcave in shape and flexible to easily pass through small capillaries and deliver oxygen to the tissues Several diseases can cause a decrease in the number of RBCs and hemoglobin content, resulting in a condition called an anemia. RBCs also may change in their size or shape when anemia is present. Note: Normal red cells in blood do not contain a nucleus. They "push out" their nucleus while still in the bone marrow to make room for the maximum amount of hemoglobin. Which cells are RBCs again? Red Blood Cells hematology_rbc.mp3 Red blood cells transport oxygen to the cells of the body. They contain hemoglobin which is responsible for carrying the oxygen. RBCs in humans are normally biconcave in shape and flexible so they can easily pass through small capillaries to deliver oxygen to the tissues. Several diseases can cause a decrease in the number of RBCs or their hemoglobin content. This condition is called anemia. RBCs also can change in size or shape when anemia is present. ? Platelets Platelets take part in the blood clotting process by: "Sticking together" and form a platelet plug at the blood vessel injury site Releasing factors that take part in the chemical coagulation process Low numbers of platelets in the blood can cause abnormal bleeding. Which cells are platelets again? Platelets hematology_platelets.mp3 Platelets are involved in the blood clotting process. They “stick together” and form a platelet plug whenever injury occurs to a blood vessel. They also release factors that take part in the chemical process of coagulation. Low numbers of platelets in the blood can result in abnormal bleeding. ? Hematology Sample Collection/Handling Blood cells are living cells and samples must be collected and handled properly to ensure accurate test results. Select the numbered steps below to learn more about special hematology sample collection and handling steps. 1 1 1 2 2 2 3 3 3 4 4 4 5 5 5 6 6 6 7 7 7 Biohazard All products or objects that come in contact with human or animal body fluids should be handled, before and after cleaning, as if capable of transmitting infectious diseases. Wear facial protection, gloves, and protective clothing. The operator should follow the recommendations to prevent the transmission of infectious agents in health-care settings as recommended by the Clinical and Laboratory Standards Institute (formerly NCCLS) in Protection of Laboratory Workers from Occupationally Acquired Infections; Approved Guideline - Third Edition. 2005. CLSI Document M29-A3. This document contains complete information on user protection and it can be used as reference. Hematology Sample Collection/Handling Baselayer: hematology_collection.mp3 Since blood cells are living cells, it is important that blood samples are collected and handled properly to protect their integrity and ensure accurate analysis test results. Select the numbered steps below to learn more about special hematology sample collection and handling steps. Slide Layer 1: hematology_collection_1.mp3 When blood is taken out of the body and collected into a tube for analysis, it normally coagulates or clots. During coagulation, the cellular components are trapped in the clot. The clot separates from the liquid portion of the blood, which is called serum. Chemistry and Immunochemistry laboratories use serum for analysis of chemical components. Slide Layer 2: hematology_collection_2.mp3 The hematology lab requires an unclotted specimen or whole blood. The tubes used in hematology are treated with an anticoagulant to prevent the blood from clotting. The anticoagulant commonly used in hematology is EDTA, or Ethylenediamine Tetraacetic acid. EDTA tubes can be identified by their purple tops. The cells remain free in the liquid portion of the anticoagulated specimen that now is called plasma and can be readily analyzed. Slide Layer 3: hematology_collection_3.mp3 It is especially important that blood is thoroughly mixed immediately after collection in an EDTA tube in order to prevent the formation of "micro-clots." These clots can cause inaccurate test results and problems in automated hematology analyzers. To mix, gently invert several times immediately after collection. Slide Layer 4: hematology_collection_4.mp3 Blood undergoes changes once it is taken from the body. The blood cells are particularly vulnerable to aging and eventual death in the sample tube. The aged and dead cells make analysis of the sample difficult or even impossible. Refrigeration of the blood slows the aging process. The blood sample should be stored in the refrigerator at 4 degrees celsius if it cannot be analyzed within 8 hours. Slide Layer 5: hematology_collection_5.mp3 Although refrigeration improves the stability of the blood cells, analysis should be completed within 8 hours of collection. A blood sample older than 24 hours, even if refrigerated, often shows signs of deterioration. A blood sample older than 48 hours will likely produce inaccurate results and one at 72 hours should not be used at all. Slide Layer 6: hematology_collection_6.mp3 A refrigerated sample must be warmed slowly to room temperature for approximately 15 minutes before analysis. Even though blood does not clot in an EDTA tube, the blood cells automatically settle to the bottom of the tube. Before analysis of the sample, the cells must be thoroughly re-suspended in the liquid plasma by gently mixing. Slide Layer 7: hematology_collection_7.mp3 In summary, use a tube with EDTA anticoagulant to collect the sample. Mix the sample gently after collection to prevent micro-clots. Analyze the sample within 8 hours if possible. Refrigerate, if needed, to minimize deterioration. And, warm the blood to room temperature and mix thoroughly before analysis. Summary Use a tube with EDTA anticoagulant to collect the sample Mix the sample gently after collection to prevent micro-clots Analyze the sample within 8 hours if possible. Refrigerate, if needed, to minimize deterioration Warm the blood to room temperature and mix thoroughly before analysis 7 Pre-Analytical Procedures Warm refrigerated blood to room temperature (approx. 15 minutes) Thoroughly re-suspend cells in the plasma by gentle mixing Tip: Test tubes can be inverted gently before analysis or mechanical rockers can be used to keep the cells suspended. Important: Vigorous mixing will harm the fragile cells of the blood. 6 Sample Aging Although refrigeration improves the stability of the blood cells: Analyze within 8 hours of collection A refrigerated sample older than 24 hours shows signs of deterioration A sample older than 48 hours will produce inaccurate results A sample older than 72 hours should not be used 5 Sample Storage Blood cells are vulnerable to aging Refrigeration slows the aging process Store the sample at 4ÕC if it cannot be analyzed within 8 hours Important: The blood sample should not be exposed to extremes in temperature. Freezing blood will totally destroy the cells and make the sample unusable. Temperatures much greater than 37ÕC also can harm the cells. 4 Sample Mixing Mix the sample immediately after collection to prevent formation of "micro-clots” Mix by gentle inversion of the sample tube several times Important: Vigorous mixing can cause damage to the cells. 3 Blood Anticoagulation Hematology analysis requires "whole" unclotted blood Sample tubes are treated with anticoagulant to prevent blood clotting Hematology commonly uses EDTA (Ethylenediamine Tetraacetic acid) as the anticoagulant Cells remain free in the liquid portion of blood, now called "plasma" 2 Blood Sample Coagulation Blood naturally coagulates or "clots" once removed from the body Cellular components are trapped within the clot The remaining liquid is called "serum" 1 ? Hematology Tests The lab processes a sample based upon the tests requested by the physician. Analysis is performed using sophisticated automated analyzers. Tests typically are ordered as a panel or profile. The two most common profiles are: Complete Blood Count (CBC) Complete Blood Count with Differential (CBC/Diff) Hematology Tests hematology_hemetests.mp3 After the blood sample is obtained, the lab will process the sample based upon the tests requested by the physician. Currently, cell counting and analysis is conducted using sophisticated automated analyzers. While hematology tests can be ordered individually, they typically are ordered as a panel or profile. The two most common profiles in hematology are the Complete Blood Count or CBC and CBC with Differential. Let’s take a look at these profiles in more detail. ? Complete Blood Count The Complete Blood Count (CBC) includes cell counts, measurement of total hemoglobin, and determination of the hematocrit and red cell indices. Select the tab arrows to learn more about the individual CBC parameters. WBC Count RBC Count Hemoglobin Hematocrit RBC Indices Platelet Count Complete Blood Count hematology_cbc.mp3 The Complete Blood Count (CBC) includes cell counts, measurement of total hemoglobin, and determination of the hematocrit and red cell indices. Select the tab arrows to learn more about the individual CBC parameters. Platelet Count The total platelet count is the number of platelets present in a specific volume of sample. Platelets are the smallest cells found in the blood. A normal platelet count is in the hundreds of thousands. Low numbers of platelets in the blood can result in abnormal bleeding High numbers of platelets often indicate a response to chronic bleeding RBC Indices RBC Indices are indicators of the average size and hemoglobin content of the cells. MCV – Mean Cell Volume is a measurement of the average size of the RBCs in the sample MCH – Mean Cell Hemoglobin is the average weight of hemoglobin inside the cells. This value is a calculation based on total Hemoglobin divided by the RBC count MCHC – Mean Cell Hemoglobin Concentration is the average concentration of hemoglobin inside the cells. The value is a calculation based upon MCV, RBC and total Hemoglobin Red cell indices are useful in differentiating different types of anemia. Hematocrit Hematocrit (Hct) is the percentage of sample volume that is taken up by the RBCs. In normal individuals, this percentage is around 45%. The remaining volume is composed of plasma, white cells and platelets. Hematocrit is associated with both the number of RBCs and their sizes. 100% 45% Hemoglobin Hemoglobin (Hgb) is the total amount of hemoglobin present in a specific volume of sample. During this analysis, sample (1) is added to reagent (2) and RBCs are lysed to release hemoglobin into the solution. The "free" hemoglobin then reacts with the reagent to produce a color (3) which is detected and measured by a colorimeter. Hemoglobin levels are closely associated with the RBC count and Hematocrit value. 1 2 3 RBC Count The total Red Blood Cell count is the number of red cells present in a specific volume of sample. They are the most numerous cells in the blood with counts in the millions. An elevated RBC count may indicate response to conditions of low oxygen such as lung disease or living in high altitude areas. Low RBCs can be due to dietary deficiencies, blood loss, conditions of red cell destruction or decreased production. RBC counts are closely associated with the Hemoglobin and Hematocrit values. WBC Count The total White Blood Cell count is the number of white cells present in a specific volume of the sample. Although WBCs are typically the largest cells found in blood, they are the least numerous with their counts running in the thousands. The WBC will increase due to infection or inflammation, or in the case of leukemia. Low WBCs may indicate bone marrow failure. ? CBC with Differential The CBC with Differential (CBC/Diff) includes the tests included in the CBC plus further classification of the white cells. It also can include immature or abnormal white cells. Select the tab arrows to learn more about WBC Differentials and additional WBC types. WBC Differential Atypical Lymphs Blasts Bands Immature Granulocytes CBC with Differential hematology_cbcdiff.mp3 The CBC with Differential or CBC/Diff includes all the tests included in the CBC but adds additional classification of the white cells called the white cell differential. The differential includes the percentages of the five normal white cell types mentioned previously but can also include immature or abnormal white cells that may be present. Select the tab arrows to learn more about WBC Differentials and additional WBC types. Immature Granulocytes An Immature Granulocyte is a neutrophil, eosinophil or basophil that is more mature than a blast but younger than a band. Immature Granulocytes can be seen in the blood in cases of severe infection and leukemia. Bands Bands are immature neutrophils with a nucleus that did not segment, typically with a "U" shaped nucleus. Bands are normal in the circulation in small numbers, but an increase in number is called a "left shift." A left shift can occur with acute infections that stimulate neutrophil production. The bone marrow may prematurely release the neutrophils before they have fully matured.   Blasts Blasts are very young WBCs. They normally are not present in the peripheral blood and do not have the ability to perform the normal immune functions of a more mature cell. Their presence in the blood is often an indication of leukemia. Atypical Lymphs An Atypical or "Reactive" Lymph is lymphocyte that has been exposed to a foreign antigen and is reacting to defend against it. It usually contains a greater amount of cytoplasm, a larger nucleus and can vary in size and shape. Increased numbers of these cells are found in the blood during a viral illnesses such as infectious mononucleosis. Select the underlined words above to identify the objects in the images. Normal Lymph Atypical Lymph 1 2 WBC Differential The WBC differential is reported as percentages of each white cell type. The total percent differential will add up to 100%. Some percentages may be misleading or inadequate. When this occurs, differential absolute numbers also are reported. Absolute numbers are obtained by multiplying the total WBC count by the percentage of each cell type The absolute neutrophil count can be used to determine if a patient has enough neutrophils to adequately fight infection. If this count is too low, the patient may be put in isolation The absolute eosinophil count may be increased in cases of allergic reactions, infections and other medical conditions Table with 3 columns and 7 rows Percent (%) Absolute # Total WBC 7,430 Neutrophils 55 4,086 Lymphocytes 36 2,675 Monocytes 5 372 Eosinophils 3 223 Basophils 1 74 ? Cell Morphology During cell counting and classification, cells also are examined to detect changes in cell morphology - cell size, shape and internal or external structures. This additional information can be helpful in both the diagnosis and treatment of disease. Cell Morphology hematology_cellmorph.mp3 During performance of the CBC or CBC/Diff, the blood cells may also be examined to detect any changes in cell morphology. Various disease conditions can cause cell size, shape and internal or external structures to change. This additional information can be helpful in both diagnosis and treatment. ? RBC Size and Shape Red cell size and shape can change when anemia is present. These morphology changes can be used to help diagnose the cause of the anemia. Microcytosis indicates the presence of very small (micro) RBCs. Microcytes will present in the peripheral blood with Iron deficiency anemia Macrocytosis indicates the presence of very large (macro) RBCs. Vitamin B12 deficiency can cause an anemia with macrocytes Anisocytosis indicates that the RBCs vary in size or are of unequal size Poikilocytosis indicates the presence of different shaped red cells. These various shapes can be caused by RBC abnormalities such as sickle cell anemia or factors that cause RBC damage such as heart valves or toxic chemicals and medications 1 3 4 2 RBC Size and Shape hematology_rbcsize.mp3 Red cell size and shape can change when anemia is present. These morphology changes can be used to help diagnose the cause of the anemia. ? RBC Hemoglobin Content Red cell hemoglobin content can change when anemia is present. These morphology changes can be used to help diagnose the cause of the anemia. Hypochromia indicates the presence of RBCs with a smaller than normal amount of hemoglobin. This causes a decrease (hypo) in their normal color (chromia) Hyperchromia indicates the presence of RBCs that contain a larger than normal amount of hemoglobin. This causes an increase (hyper) in their normal color (chromia) when examined. The increase in hemoglobin could also cause these cells to be round instead of biconcave in shape Nucleated RBCs are red cells that are very immature, are still forming hemoglobin, and still contain a nucleus. They are present in the bone marrow, and normally not seen in peripheral blood 1 2 3 RBC Hemoglobin Content hematology_rbchgb.mp3 Red cell hemoglobin content can change when anemia is present. These morphology changes can be used to help diagnose the cause of the anemia. ? WBC Morphology White cells undergo changes in their internal components during infection, inflammation or malignancy. There are many internal changes that can occur but some of the more common ones are: Toxic Granulation indicates the presence of large dark granules in the cytoplasm of neutrophils, associated with severe infection, chemical poisoning, and other toxic states Auer Bodies are unique, pink or red rod-shaped inclusions that are seen in the cytoplasm of blasts in patients with a specific type of acute leukemia called Acute Myelogenous Leukemia (AML) Vacuoles are large clear areas within the cytoplasm of the cell that can be seen in cases of severe infection 1 2 3 WBC Morphology hematology_wbcmorph.mp3 White cells undergo changes in their internal components during infection, inflammation or malignancy. There are many internal changes that can occur but some of the more common ones are toxic granulation, auer bodies and vacuoles. ? Platelet Morphology Platelets are normally the smallest cell in the blood and are about one tenth the size of RBCs. They can change in size, shape and internal components in various disease conditions. Large platelets can be seen when there is a high rate of production or abnormal platelet production Platelet clumping is typically an artifact of blood collection 1 2 Platelet Morphology hematology_pltmorph.mp3 Platelets are normally the smallest cell in the blood and are about one tenth the size of RBCs. They can change in size, shape and internal components in various disease conditions. ? Manual Smear Review and Differential Automated hematology analyzers produce flags when a cell morphology change is detected or immature/abnormal cells are suspected. A manual blood smear is made, stained and reviewed to confirm the abnormal results produced by the automated analyzer. Select the numbered steps below to learn more about the manual blood smear review and differential. 1 1 1 2 2 2 3 3 3 4 4 4 5 5 5 6 6 6 Manual Smear Review and Differential Baselayer: hematology_manualsmear.mp3 Many automated hematology analyzers produce flags when cell morphology changes or immature or abnormal cells are suspected. If a flag occurs, the hematology technologist will often make a blood smear of the sample, stain it, and examine it under the microscope. Based upon laboratory policy, the technologist will confirm the automated results, add the manual smear review results to the automated results, and will sometimes also perform a manual differential to validate the automated diff. Select the numbered steps below to learn more about the manual blood smear review and differential. Layer 1: hematology_manualsmear_1.mp3 The manual smear review and diff procedure begins with blood smear preparation. A wedge smear is prepared by placing a drop of blood onto a slide and smearing it using another slide or smearing device. This smear can be prepared either automatically by the hematology analyzer or manually by the instrument operator. Layer 2: hematology_manualsmear_2.mp3 Next, the slide is stained in order to better visualize the cells. The stains used in most labs are Wright's or Wright's-Giemsa which stain the cells by using blue and red dyes. Staining can be done manually but is usually done on instruments that are semi or fully automated. During the staining process, the cells are first stained, then buffered and rinsed. The slide must be allowed to dry before it is examined under a microscope Layer 3: hematology_manualsmear_3.mp3 Once the slide is prepared and stained, it is ready for microscopic evaluation. The slide is placed on the microscope and examined at 100 times magnification. The feathered edge is located at the end of the slide. In this area the cells are separated from each other and are able to be counted and classified more accurately. The feathered edge of the slide is examined following an organized pattern. During the slide examination the WBCs, RBCs, and Platelets are evaluated for abnormal morphology. Layer 4: hematology_manualsmear_4.mp3 For the manual differential, 100 WBCs are counted and classified into their various types. Layer 5: hematology_manualsmear_5.mp3 Results from the automated analyzer are confirmed and additional manual findings are added to the final report. Layer 6: hematology_manualsmear_6.mp3 In summary, prepare a wedge blood smear. Stain the blood smear. And examine the smear microscopically and perform a manual differential. Summary Prepare wedge blood smear Stain blood smear Examine smear microscopically and perform manual differential 6 Report Results Automated results are confirmed Additional manual findings are added 5 Perform Manual Differential One hundred WBCs are counted and classified into their various types to perform the WBC differential. 4 Examine Blood Smear The slide is now ready for microscopic evaluation at 100X magnification. The "feathered edge" of the slide is examined following an organized pattern WBCs, RBCs and Platelets are evaluated for abnormal morphology 3 1 2 Stain Blood Smear The slide is stained to better visualize the cells. Stains commonly used are Wright's or Wright's-Giemsa which stain the cells by using blue and red dyes. Staining can be performed manually or on semi or fully automated instruments. During the procedure, the cells are: Stained Buffered Rinsed Dried 2 Prepare Wedge Blood Smear Place a drop of blood onto a slide Smear it using another slide or smearing device Preparation can be done automatically by the hematology analyzer or manually by the instrument operator. 1 ? Reticulocyte Count Reticulocytes (retics) are young red cells that are still maturing. A special stain must be used to detect the retics present within the circulating RBC population. Their numbers in blood indicate the level of new red cell production by the bone marrow. Normal is 1% to 2%. Retic counting can be performed manually or by an automated analyzer. 1 1 1 2 2 2 3 3 3 4 4 4 5 5 5 Select the underlined word above to identify the object in the image and select the numbered steps below to learn more about reticulocyte counts. Reticulocyte Count hematology_retic.mp3 Another test that is commonly performed in hematology is the reticulocyte count. Reticulocytes, or retics, are young red cells that are still maturing. A special stain must be used to detect the retics present within the circulating RBC population. Their number indicates the level of new red cell production by the bone marrow. A normal count is about 1 to 2 percent. Retic counting in the hematology laboratory is either done manually or by an automated analyzer. Select the underlined word above to identify the object in the image and select the numbered steps below to learn more about reticulocyte counts. Reporting Results The Retic Count then is reported a percentage of the total RBC count with 1.0% to 2.0% being normal. 5 Counting Using a counting device, the technologist manually examines the slide to count 1000 RBCs and simultaneously enumerate the number of retics. Retics are recognized by their blue granules. 4 Microscopic Evaluation The slide is placed on the microscope and read at 100X magnification. The feathered edge of the slide is examined. 3 Smear Preparation After staining is complete, a wedge smear is prepared from the mixture. 2 Staining Blood is mixed with a special stain which differentiates between retics and more mature red cells. Only retics pick up the stain because of the RNA present inside the cell. 1 Knowledge Check You have just learned about Hematology sample collection, handing, and tests. Select Start to test your knowledge of the presented content. Start ? Knowledge Check Select the best answer. ? Question 1 of 3 What is the purpose of the bone marrow? Produce blood cells Participate in blood clotting Participate in the immune response Transport oxygen to tissues of the body Multiple Choice Incorrect RBCs transport oxygen from the lungs to the body tissues. Incorrect WBCs participate in the immune response and defend the body against invasion by foreign substances. Incorrect Platelets participate in blood clotting. They stick together form a platelet plug. Correct Select the best answer. ? Question 2 of 3 What is the main purpose of the CBC/Diff? Count the cell types and classify the white cells Count the cell types Determine cell size, shape and structure changes Determine the number of young RBCs Multiple Choice Incorrect The reticulocyte count determines the percentage of young RBCs in the blood. Incorrect Cell morphology is evaluated to determine cell size, shape, and internal or external structures changes. Incorrect The CBC includes cell counts, hemoglobin, hematocrit and red cell indices. Correct Select the best answer. ? Question 3 of 3 Why is EDTA used in hematology? Prevent clotting of the sample Minimize cell deterioration Prevent formation of "micro-clots” Analyze the serum Multiple Choice Incorrect Chemistry and Immunochemistry laboratories use serum for analysis of chemical components. Incorrect Immediate sample mixing after collection prevents formation of "micro-clots”. Incorrect Blood is refrigerated to minimize cell deterioration. Correct Knowledge Check Completion YOUR SCORE: PASSING SCORE: Review Retry Retry Continue Continue %Results.ScorePercent%% %Results.PassPercent%% ? You have just completed the Knowledge Check. Select Review to assess how your responses compare to the correct answers. Select Retry to test your knowledge again or select Continue to advance through the course. Assessment Results ? Automated Hematology Analyzers Sophisticated automated instruments are used in the hematology laboratory. Mechanisms are often included to: Mix the sample tube Puncture the cap to withdraw sample Create a stained blood smear as needed Automated Hematology Analyzers hematology_automated.mp3 Hematology laboratories typically rely on sophisticated automated instruments to analyze hematology samples. These analyzers incorporate mechanisms to mix the sample and then puncture the cap of the sample tube to withdraw a sample for analysis. During analysis, some analyzers also have the ability to make a stained blood smear for manual microscopic review as needed. ? Analytical Techniques Hematology analyzers incorporate specialized analytical techniques to count, classify and examine cells including: Colorimetry Electrical Impedance Optical Counting Cytochemical Staining Flow Cytometry Analytical Techniques hematology_analytical.mp3 Hematology analyzers incorporate specialized analytical techniques to count the cells, classify them into the different the cell types and examine both their internal and external characteristics. Some of the most common techniques used are colorimetry, electrical impedance, optical counting, cytochemical staining and flow cytometry. ? Colorimetry Colorimetry is used in the hematology laboratory to measure Total Hemoglobin. In this test method: Blood is mixed with a reagent RBCs lyse and release the hemoglobin Hemoglobin reacts with the reagent to produce a color that is measured in a colorimeter Select the colored callouts below to learn more about colorimetry. Colorimetry hematology_colorimetry.mp3 Colorimetry is used in hematology to measure Total Hemoglobin. In this method, a sample of blood is added to a reagent in a test tube or other container. The reagent causes the RBCs to lyse and release the hemoglobin from within. The hemoglobin then reacts with the reagent to produce a color that is measured using a device called a colorimeter. Select the colored callouts to learn more about colorimetry. Readout Device The readout device receives the signal and records it as either absorbance (A) or percent transmittance (%T). This value then is converted to the total hemoglobin result using computer software and calibration data. Electrical Signal The electrical signal is processed and sent to a readout device. Detector A photodetector detects the amount of light exiting from the cuvette and converts it to an electrical signal. Transmitted Light This light exits after passing through the reaction mixture in the cuvette. Light that does not exit has been absorbed by the reaction mixture. Cuvette This container holds the blood/reagent reaction mixture during the measurement. The light shining through the reaction mixture is absorbed depending upon the amount of reaction end-product formed. Monochromatic Light The single wavelength of light also is called the incident light since it is the light present before it passes through the reaction mixture. Wavelength Selector A wavelength selector is used to isolate a specific wavelength of light. For hemoglobin measurement, a wavelength of 540 nm is used. White Light White light is composed of many different wavelengths of light from ultraviolet through the visible color spectrum up to infrared. Light Source A light source, often a tungsten halogen lamp, is used to produce light. ? Electrical Impedance One method of counting and classifying cells in hematology is electrical impedance. Impedance technology uses an electrical current that passes through an aperture (4) between two electrodes (5) Cells pass through the aperture, block the electrical current, and create resistance Resistance produces a signal or a pulse (6) whose height is proportional to cell size Electronic thresholds are used to separate the cells into categories dependent upon size Start 1 2 3 4 6 5 5 Electrical Impedance hematology_electric.mp3 A common method of counting and classifying cells in hematology is electrical impedance. Impedance technology utilizes an electrical current that passes through an aperture between two electrodes. When a cell passes through the aperture, it blocks the electrical current and creates resistance. Resistance produces a signal or a pulse. The amount of resistance or the height of the pulse is proportional to the cell size. Electronic thresholds are then used to separate the cells into categories dependent upon their size. Select the Start button to learn more about electrical impedance. Observe the different cells in the video illustration to see how they affect the pulse height. Platelet RBC WBC ? In Optical counting, a beam of light is directed through a flowcell (4). If there are no cells in the flowcell, the darkstop (5) prevents the light from reaching the detector (6). A cell within the flowcell scatters the light around the darkstop The scattered light reaches the detector which counts the cell The amount of light scatter is directly proportional to the size of the cell Optical Counting Select the play button in the video illustration to observe how the different cells affect the light scatter. Platelet RBC WBC Note: Measurement of scattered light at different angles can be used to determine internal characteristics of the cells. Analyzers capture specific angles of light scatter and use complex algorithms to determine; for example, differences in cell morphology or the presence of abnormal cells. Optical Counting hematology_optical.mp3 Another way that cells can be both counted and analyzed is by optical counting. A beam of light is directed through a flowcell. If there are no cells in the flowcell, the light is prevented from reaching the detector by the darkstop. When a cell is introduced into the flowcell it scatters the light around the darkstop. When the detector sees the scattered light, it counts a cell. The amount of light scatter is directly proportional to the size of the cell. Select the play button in the video illustration to observe how the different cells affect the light scatter. ? Cytochemical Staining Cytochemical staining differentiates between cells that appear identical in physical structure. These stains react with specific chemical components within cells (1). The reaction produces changes that cause the cell to absorb light as it passes through an optical counting area. The amount of light absorbed by the cell is used to classify the cell. Select the colored callouts below to learn more about cytochemical staining detection. Note: Fluorescent dyes also can be used that first absorb the light and then re-emit it at a different wavelength. 1 Cytochemical Staining hematology_cytochemical.mp3 Cytochemical staining helps to differentiate between cells that appear identical in physical structure. Cytochemical stains react with specific chemical components within the cells. The reaction produces changes in the cell that cause it to absorb light as it passes through an optical counting area. The amount of light absorbed by the cell is then converted to a reading used to classify the cell. Select the colored callouts to learn more about cytochemical staining detection. Readout Device The readout device receives the signal and processes it for classification by the analyzer computer software. Electrical Signal The electrical signal is processed and sent to a readout device. Detector A photodetector detects the amount of light exiting the flowcell and converts it to an electrical signal. Transmitted Light Transmitted light is the light that exits from the flowcell. Light that does not exit has been absorbed by the cell. Cell A cytochemically stained cell passing through the beam absorbs the light. One that is not stained does not absorb the light. Optical Flowcell The optical flowcell allows cells to pass one-by-one through the incident light beam. Wavelength Selector A wavelength selector is used to isolate a specific wavelength of light. Monochromatic Light The single wavelength of light is also called the incident light since it is the light present before it passes through the optical flowcell. Light Source A light source, often a tungsten halogen lamp, is used to produce light. ? Flow cytometry enhances both impedance and optical counting technologies. It uses hydrodynamic focusing to narrow and focus the flow of a stream of sample liquid by: Utilizing stream of liquid called a sheath Injecting the sample stream into the middle of the flowing sheath The fluids differ in velocity or density so that they do not mix. Single cells in the sample fluid pass by the detection area, independent from each other to be counted and analyzed. Flow Cytometry Flow Cytometry hematology_flowcyto.mp3 Flow cytometry is used to enhance both impedance and optical counting technologies and produce more accurate and precise results. It uses a technique called hydrodynamic focusing to narrow and focus the flow of a stream of sample liquid by surrounding it with another stream of liquid, called a sheath. The sample stream is injected into the middle of the flowing sheath fluid. The two fluids differ in their velocity or density so they do not mix. Single cells in the sample fluid can then pass by the detection area independent from each other to be counted and analyzed. ? Course Review Congratulations. You have completed the Hematology Basic Overview Online Training course. Select the numbered buttons below to review the material before proceeding to the final assessment. Identify analytical methods used in the hematology lab Recognize common tests performed in hematology Recognize procedures for the proper handling of hematology specimens Identify the different blood cells and their functions 1 1 2 2 2 3 3 3 4 4 4 Course Review Identify analytical methods used in the hematology lab Colorimetry is used to measure total Hemoglobin by mixing together blood and a reagent and detecting the color change created by the chemical reaction Electrical Impedance is used to count and classify blood cells by detecting an electrical current change that is caused as a cell passes through an aperture Optical Counting uses a light directed at cells passing through a flowcell. As cells scatter the light they are counted and their size and internal characteristics can be determined Cytochemical Staining is used to stain specific chemical components of cells to help differentiate between cells that appear identical in physical structure Flow Cytometry is used to enhance impedance and optical counting by hydrodynamically focusing a stream of cells as they pass through the detection areas Recognize common tests performed in hematology The Complete Blood Count (CBC) includes WBC, RBC and Platelets counts, total Hemoglobin, Hematocrit, and Red Cell Indices The CBC/Diff includes all tests of the CBC plus a categorization of the different types of normal or abnormal WBCs found in the sample Cell Morphology includes descriptions of abnormal size, shape or internal and external structures of the cells Manual Smear Review/Differential is done to confirm automated results and flags and add additional detail about abnormal cells and morphology The Reticulocyte Count is a special test that is used to detect the presence of young RBCs in the blood Recognize procedures for the proper handling of hematology specimens Collect samples in EDTA anticoagulated tubes and mix gently immediately after collection to prevent micro-clots Analyze samples within 8 hours or refrigerate for up to 48 hours Warm samples to room temperature and mix gently before analysis Identify the different blood cells and their functions All the blood cells originate in the bone marrow. White Blood Cells (WBCs) are part of the immune system and defend the body against invasion by foreign organisms or chemicals. There are five types WBCs normally found in the blood: Neutrophils, Lymphocytes, Monocytes, Eosinophils and Basophils Red Blood Cells (RBCs) are responsible for transporting oxygen to the body tissues. They contain hemoglobin which carries the oxygen Platelets take part in the blood clotting process by forming platelet plugs and releasing factors in the coagulation process Disclaimer Please note that the learning material is for training purposes only. For the proper use of the software or hardware, please always use the Operator Manual or Instructions for Use (hereinafter collectively “Operator Manual”) issued by Siemens Healthineers. This material is to be used as training material only and shall by no means substitute the Operator Manual. Any material used in this training will not be updated on a regular basis and does not necessarily reflect the latest version of the software and hardware available at the time of the training. The Operator Manual shall be used as your main reference, in particular for relevant safety information like warnings and cautions. Please note: Some functions shown in this material are optional and might not be part of your system. Certain products, product related claims or functionalities (hereinafter collectively “Functionality”) may not (yet) be commercially available in your country. Due to regulatory requirements, the future availability of said Functionalities in any specific country is not guaranteed. Please contact your local Siemens Healthineers sales representative for the most current information. The reproduction, transmission or distribution of this training or its contents is not permitted without express written authority. Offenders will be liable for damages. All names and data of patients, parameters and configuration dependent designations are fictional and examples only. All rights, including rights created by patent grant or registration of a utility model or design, are reserved. © Siemens Healthcare GmbH 2022 Siemens Healthineers Headquarters\Siemens Healthcare GmbH\Henkestr. 127\ 91052 Erlangen, Germany\Telephone: +49 9131 84-0\siemens-healthineers.com ? Disclaimer Assessment This assessment will test your retention of the presented content. A passing score of 80% or higher is required to complete the course and earn your certificate. You may repeat the assessment as many times as needed. Start ? Assessment Select the best answer. ? Question 1 of 10 Your friend told you that blood testing indicated she had a very low RBC count. Based upon your knowledge of hematology you predict: She probably has problems with physical tasks and tires quickly She is also running a fever and likely has an infection She must be very careful not to cut herself because she has an increased tendency to bleed She probably has problems sleeping and should take medication Multiple Choice Incorrect Problems with sleeping are not related to a low RBC count. Incorrect A low platelet count would increase the tendency to bleed. Incorrect An elevated WBC count would indicate an infection with fever. Incorrect Select the best answer. ? Question 2 of 10 A patient that has bone marrow failure due to drug chemotherapy would likely show CBC results with: Low WBC, RBC and Platelet counts High WBC, RBC, and Platelet counts Normal WBC, RBC, and Platelet counts A high WBC count and low RBC and Platelet counts Multiple Choice Incorrect Bone marrow failure would affect all the cells similarly. Incorrect Bone marrow failure would cause the counts to change from their normal values. Incorrect Bone marrow failure would not cause these counts to be elevated. Correct Select the best answer. ? Question 3 of 10 It is important to gently mix hematology samples in order to: Prevent micro-clots from forming and re-suspend the cells before analysis Allow the cellular components to be trapped within the clot Keep the red cells stable and slow the deterioration of the white cells Prevent cell metabolism Multiple Choice Incorrect Mixing does not prevent cell metabolism. Incorrect Refrigeration, not mixing, helps stabilize cells and slow their deterioration. Incorrect Anticoagulant prevents the blood from clotting. Correct Select the best answer. ? Question 4 of 10 It is important to try to analyze hematology samples within _________ hour(s) of collection to ensure the most accurate results. 8 1 24 48 Multiple Choice Incorrect It is best not to use samples 48 hours old since they will show signs of deterioration. Incorrect It is better to analyze hematology samples within a shorter time period than 24 hours, especially if the sample cannot be refrigerated. Incorrect Blood samples older than 1 hour can still be analyzed with accuracy. Correct Select the best answer. ? Question 5 of 10 Your child has a fever, sore throat, cough, and muscle aches for four days. You visit the doctor who orders some blood tests. The hematology test most likely to be ordered to diagnose and treat your infection is: CBC/Diff CBC Retic Count Total Hemoglobin Multiple Choice Incorrect The total hemoglobin is used for diagnosing and treating anemia, not infection. Incorrect The retic count is used to estimate bone marrow activity and is not used in diagnosing and treating infections. Incorrect The CBC will provide good information, but another test is even more diagnostic in cases of infection. Correct Select the best answer. ? Question 6 of 10 A patient blood with low RBC count, Total Hemoglobin, and Hematocrit will also likely show abnormal: Red cell morphology White cell morphology Platelet morphology Bone morphology Multiple Choice Incorrect Bone morphology is not related to low red cell test values. Incorrect These test results would not affect Platelet morphology. Incorrect These test results would not affect WBC morphology. Correct Select the best answer. ? Question 7 of 10 A patient CBC/Diff report from an automated analyzer shows an abnormal WBC differential and flags for abnormal RBC morphology. The hematology technologist will likely: Perform a manual diff and smear review Order another sample from the patient Refrigerate the sample and analyze it again Ask the doctor to order a bone marrow analysis Multiple Choice Incorrect Abnormal CBC/Diff results to not warrant ordering a bone marrow analysis. Incorrect A refrigerated sample should first be warmed to room temperature before analysis. Incorrect There is another step the technologist should first perform before asking for another sample. Correct Select the best answer. ? Question 8 of 10 Analysis of Total Hemoglobin is performed in the hematology laboratory using: Colorimetry Electrical Impedance Optical Counting Cytochemical Staining Multiple Choice Incorrect Cytochemical staining is used to differentiate cells with similar physical characteristics. Incorrect Optical counting is used to count and classify cells. Incorrect Electrical impedance is used to count and classify cells. Correct Select the best answer. ? Question 9 of 10 Cytochemical staining is used in hematology to: Differentiate between cells with similar physical structure Prepare the cells for a manual smear review and differential Enhance impedance and optical counting technologies Multiple Choice Incorrect Flow cytometry is used to enhance impedance and optical counting technologies. Incorrect A different staining technique is used to prepare slides for manual smear review and differential. Correct Select the best answer. ? Question 10 of 10 Flow cytometry uses a(n) ___________ fluid to focus the flow of cells past a detector. Sheath Sample Colored Multiple Choice Incorrect Color is not important in flow cytometry. Incorrect The sample fluid is what is focused by the other fluid used in flow cytometry. Correct Assessment Results YOUR SCORE: PASSING SCORE: Review Retry Retry Continue Continue Continue %Results.ScorePercent%% %Results.PassPercent%% ? Assessment Results You did not pass the course. Take time to review the assessment then select Retry to continue. Congratulations. You passed the course.. Exit To access your Certificate of Completion, select the Launch button drop down on the course overview page. You can also access the certificate from your PEPconnect transcript. ? You have completed the Hematology Basic Overview Online Training. Completion Navigation Help Select the icon above to open the table of contents. Click Next to continue. Next Welcome Slide The timeline displays the slide progression. Slide the orange bar backwards to rewind the timeline. Click Next to continue. Next Timeline Select the CC icon to display closed captioning (subtitles). Click Next to continue. Next Caption Icon add subtitles Select the buttons to learn more about a topic. Be sure to review all topics before navigating to the next slide. Click Next to continue. Next Tab Arrow Slide Select the X to close the pop-up. Click Next to continue. Next Layer Slide Select Submit to record your response. Click the X in the upper right corner to exit the navigation help. Assessment Slide Question Bank 1 HILS2242 | Effective Date: 14-JAN-2022 1.1 Welcome 1.2 What is Hematology? 1.3 Parts of a Cell 1.4 Bone Marrow 1.5 White Blood Cells 1.6 Red Blood Cells 1.7 Platelets 1.8 Hematology Sample Collection/Handling 1.9 Hematology Tests 1.10 Complete Blood Count 1.11 CBC with Differential 1.12 Cell Morphology 1.13 RBC Size and Shape 1.14 RBC Hemoglobin Content 1.15 WBC Morphology 1.16 Platelet Morphology 1.17 Manual Smear Review and Differential 1.18 Reticulocyte Count 1.19 Knowledge Check 1.24 Automated Hematology Analyzers 1.25 Analytical Techniques 1.26 Colorimetry 1.27 Electrical Impedance 1.28 Optical Counting 1.29 Cytochemical Staining 1.30 Flow Cytometry 1.31 Course Review 1.32 Disclaimer 1.33 Assessment