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Hematologic ExaminationsPrepared by: Group 6 (Chito, Chua, Co, Cortez)
Hemoglobin Determination
Hemoglobin Determination
• Determines the amount of hemoglobin in blood, consequently measuring amount of oxygen
• Used to detect:
1. Fat embolism
2. Iron deficiency anemia
3. Lactic acidosis
4. Sickle cell disease
Hemoglobin Determination
• Measured using a hemoglobinometer
• Procedure:
Source: www.medicine.mcgill.ca/physio/vlab/bloodlab/hemoglobin_n.htm
Hemoglobin Determination
1.Drop of blood is placed on one side of the chamber glass
Hemoglobin Determination
2. Blood is stirred with a hemolysis applicator, with saponin
Hemoglobin Determination
3. Cover glass is placed on top of chamber glass.4. Both are slid onto metal clip.
Hemoglobin Determination
5. Blood chamber is inserted into a slot in the hemoglobinometer.
Hemoglobin Determination
• When the light switch button is pressed, a green split field appears
• Take the reading the arrow is pointed at when the two halves are of the same shade of green
Hemoglobin Determination
• Another method is the cyanmethemoglobin method
• Blood is mixed with Drabkin’s sol’n
• Ferricyanide in the sol’n oxidizes iron – methemoglobin
• Methemoglobin + cyanide -> cyanmethemoglobin
• Color is measured
Hemoglobin Determination
• Two manual methods: Haden Hausse and Sahli-Hellige
• Blood + dilute HCl
• Hemoglobin -> brownish-colored acid hematin
• Acid hematin sol’n is compared with a color standard
Hematocrit Determination
Hematocrit Determination
• Indicator of Blood Viscosity1
• Measures the percentage of red blood cells in whole blood2
• Also called Packed-cell Volume2
Source: 1= Baldeo, Biendima, Go, Olivar, Soriano. Calorimetric Determination of Hemoglobin. Retrieved from: http://www.slideshare.net/JayEdneilOlivar/hemoglobin-and-hematocrit-determination2= Tpub. Hematocrit Determination. Retrieved From: http://www.tpub.com/corpsman/234.htm3=UTMB. Hematocrit Determination. Retrieved From: http://shp.utmb.edu/cls/NursePractitioners/hct.pdf
Hematocrit Determination
• The microhematocrit determination is based on the principle that if a sample of whole (anticoagulated) blood is centrifuged for a period of time and at a speed to achieve maximum packing of the cells, the height of the red cell column divided by the height of the column of cells and plasma represents the total volume of whole blood that is occupied by the erythrocytes3.
Source: 1= Baldeo, Biendima, Go, Olivar, Soriano. Calorimetric Determination of Hemoglobin. Retrieved from: http://www.slideshare.net/JayEdneilOlivar/hemoglobin-and-hematocrit-determination2= Tpub. Hematocrit Determination. Retrieved From: http://www.tpub.com/corpsman/234.htm3=UTMB. Hematocrit Determination. Retrieved From: http://shp.utmb.edu/cls/NursePractitioners/hct.pdf
Hematocrit Determination
• Source: https://www.youtube.com/watch?v=FebAWEgOydM
Hematocrit Determination• Interpretation1
Normal hematocrit values are: – Newborn: up to 60%– Adults: (males): 40- 54% – (Females): 36 – 46%
• Pregnancy: decreased hematocrit, especially in the last trimester as plasma volume increases
• Children: varies with age • Hematocrit = 3x hemoglobin for normal RBC count and
hemoglobin amount onlySource: 1= Baldeo, Biendima, Go, Olivar, Soriano. Calorimetric Determination of Hemoglobin. Retrieved from: http://www.slideshare.net/JayEdneilOlivar/hemoglobin-and-hematocrit-determination2= Tpub. Hematocrit Determination. Retrieved From: http://www.tpub.com/corpsman/234.htm3=UTMB. Hematocrit Determination. Retrieved From: http://shp.utmb.edu/cls/NursePractitioners/hct.pdf
Hematocrit and Blood Viscosity1
Source: 1= Baldeo, Biendima, Go, Olivar, Soriano. Calorimetric Determination of Hemoglobin. Retrieved from: http://www.slideshare.net/JayEdneilOlivar/hemoglobin-and-hematocrit-determination2= Tpub. Hematocrit Determination. Retrieved From: http://www.tpub.com/corpsman/234.htm3=UTMB. Hematocrit Determination. Retrieved From: http://shp.utmb.edu/cls/NursePractitioners/hct.pdf
Abnormal Hematocrit Values1
• Elevated hematocrit– Sign of dengue shock syndrome.– Polycythemia vera – bone marrow produces
excessive numbers of RBCs– Chronic obstructive pulmonary diseases– Hypoxia– Capillary leak syndrome– Dehydration (hypovolemia)
Source: 1= Baldeo, Biendima, Go, Olivar, Soriano. Calorimetric Determination of Hemoglobin. Retrieved from: http://www.slideshare.net/JayEdneilOlivar/hemoglobin-and-hematocrit-determination2= Tpub. Hematocrit Determination. Retrieved From: http://www.tpub.com/corpsman/234.htm3=UTMB. Hematocrit Determination. Retrieved From: http://shp.utmb.edu/cls/NursePractitioners/hct.pdf
Abnormal Hematocrit Values1
• Decreased Hematocrit– Hemorrhage– Chronic kidney disease– Inadequate iron uptake– Iron deficiency due to blood loss during menstruation– Anemia– Malnutrition– Overhydration
Source: 1= Baldeo, Biendima, Go, Olivar, Soriano. Calorimetric Determination of Hemoglobin. Retrieved from: http://www.slideshare.net/JayEdneilOlivar/hemoglobin-and-hematocrit-determination2= Tpub. Hematocrit Determination. Retrieved From: http://www.tpub.com/corpsman/234.htm3=UTMB. Hematocrit Determination. Retrieved From: http://shp.utmb.edu/cls/NursePractitioners/hct.pdf
Abnormal Hematocrit Values1
• A hematocrit of less than 15% can result in cardiac failure
• A hematocrit of over 60% may result in spontaneous blood clotting
Source: 1= Baldeo, Biendima, Go, Olivar, Soriano. Calorimetric Determination of Hemoglobin. Retrieved from: http://www.slideshare.net/JayEdneilOlivar/hemoglobin-and-hematocrit-determination2= Tpub. Hematocrit Determination. Retrieved From: http://www.tpub.com/corpsman/234.htm3=UTMB. Hematocrit Determination. Retrieved From: http://shp.utmb.edu/cls/NursePractitioners/hct.pdf
References
• Source: Calorimetric Determination of Hemoglobin. Retrieved from:
http://www.slideshare.net/JayEdneilOlivar/hemoglobin-and-hematocrit-determination
Hematocrit Determination. Retrieved From: http://www.tpub.com/corpsman/234.htm
Hematocrit Determination. Retrieved From: http://shp.utmb.edu/cls/NursePractitioners/hct.pdf
Complete Blood Count (CBC)
Normal Values
Source: Mercy Medical Center- North Iowa. (n.d.). CBC Normal Ranges. Retrieved from http://www.mercynorthiowa.com/cbc-normal-ranges
HEMOGLOBIN (G/DL)AGE MALE FEMALE> 65 years 12.6 - 17.4 11.7 - 16.14 5 - 64 years 13.1 - 17.2 11.7 - 16.018 - 44 years 13.2 - 17.3 11.7 - 15.512 - 17 years 11.7 - 16.6 11.5 - 15.39 - 11 years 12.0 - 15.0 12.0 - 15.06 months - 8 years 11.2 - 14.1 11.2 - 14.14 - 5 months 10.3 - 14.1 10.3 - 14.12 - 3 months 9.4 - 13.0 9.4 - 13.01 month 10.7 - 17.1 10.7 - 17.114 - 30 days 13.4 - 19.8 13.4 - 19.80 - 13 days 13.5 - 20.5 13.5 - 20.5
• Lower than normal Hbg may be due to:
1. Anemia
2. Bleeding from GIT or bladder
3. Chronic kidney disease
4. Stopped erythropoiesis
5. Poor nutrition
6. Low level of iron, folate, vitamin B12, B6
7. Other chronic illnesses
• Higher than normal Hbg may mean:
1. Hypoxia – Birth defects– Failure of the right side of the heart– Severe COPD– Pulmonary fibrosis
2. Polycythemia vera
3. Dehydration
References
Thomson Micromedex (1984). Retrieved from http://www.mc.vanderbilt.edu/diglib/consumerhealth/labs/49/150149.htm
The McGill Physiology Virtual Lab (Aug 15, 2008) Retrieved from www.medicine.mcgill.ca/physio/vlab/bloodlab/
hemoglobin_n.htm
Integrated Publising, Inc. (Aug 31, 2014). Retrieved from http://www.tpub.com/corpsman/233.htm
Mercy North Iowa (2014). Retrieved from http://www.mercynorthiowa.com/cbc-normal-ranges
MedlinePlus (Aug 15, 2014) Retrieved from http://www.nlm.nih.gov/medlineplus/ency/article/003645.htm
Platelet Count
Platelet Count
• A test used to detect low or high number of platelets in the blood
• Included in the CBC
• Recall: platelets are important in hemostasis
Source: American Association for Clinical Chemistry. (2014.). Platelet count. Retrieved from http://labtestsonline.org/understanding/analytes/platelet/tab/test/a
Platelet Count
• More difficult to perform than WBC and RBC Count because:– Very small– Tendency to clump together
Source: Department of Medical Microbiology. (2014). A manual of clinical laboratory diagnostic procedures. College of Public Health: UP Manila
Platelet Count
Uses of the Test
• Investigate bleeding disorders
• Assess clotting ability
• Monitor drug treatments
Source: Department of Medical Microbiology. (2014). A manual of clinical laboratory diagnostic procedures. College of Public Health: UP Manila
Platelet Count
2 Methods of Platelet Counting
1.Direct Platelet Count
2.Indirect Platelet Count
Source: Department of Medical Microbiology. (2014). A manual of clinical laboratory diagnostic procedures. College of Public Health: UP Manila
Direct Platelet Count
Source: Department of Medical Microbiology. (2014). A manual of clinical laboratory diagnostic procedures. College of Public Health: UP Manila
Draw diluting fluid (Rees and Ecker) up and down the capillary stem
Shake the RBC pipette fo 3 minutes
Count the platelets in large central square using the HPO.
Direct Platelet Count
Source: Department of Medical Microbiology. (2014). A manual of clinical laboratory diagnostic procedures. College of Public Health: UP Manila
Calculation:
Platelet count (/mm3)= N x dilution factor x depth factorArea
Example: 200 cells counted Platelet count (/mm3)= N x dilution factor x depth factor
Area Platelet count (/mm3)= 200 x 200 x 10
1 Platelet count (/mm3)= 400,000/mm3
Indirect Platelet Count
Source: Department of Medical Microbiology. (2014). A manual of clinical laboratory diagnostic procedures. College of Public Health: UP Manila.
Make a blood smear from venous blood.
Using the OIO, count the number of platelets/1,000 RBC.
www.ruf.rice.edu
Indirect Platelet Count
Indirect Platelet Count
Source: Department of Medical Microbiology. (2014). A manual of clinical laboratory diagnostic procedures. College of Public Health: UP Manila
Calculation:
Platelet count (/mm3)= P x RBCs per mcL 1,000
Example: 60 cells counted/1,000 RBCs; RBC= 5.0x10^12/L Platelet count (/mm3)= P x RBCs per mcL
1,000 Platelet count (/mm3)= 60 x 5,000,000
1,000 Platelet count (/mm3)= 300,000
Indirect Platelet Count
Source: Mercy Medical Center- North Iowa. (n.d.). CBC Normal Ranges. Retrieved from http://www.mercynorthiowa.com/cbc-normal-ranges
Normal Values:
PLATELETS K/MCL
AGE MALE FEMALE
All ages 150 - 440 150 - 440
Indirect Platelet Count
Notes:•A low or borderline platelet count obtained by the indirect method should be checked carefully by a count using a direct method.•The indirect method is not recommended as a method of choice.
The indirect platelet count is higher than the direct count because the red cells, which are used as a point of reference in the indirect method, are not randomly distributed beneath the coverslip. The red cells are concentrated at the edge of the coverslip so that the true ratio of red cells to platelets cannot be accurately established. Indirect platelet counts based on the ratio in the central areas of the coverslip are too high.
Source: Fitch, H. (1957). An explanation of the discrepancy between direct and indirect platelet counts. Blood: 12(7). Retrieved from http://www.bloodjournal.org/content/12/7/671?sso-checked=true
Causes of Abnormal Values
TEST LOW COUNT HIGH COUNT
Platelet Count
Known as thrombocytopenia:Viral infection Rocky mountain spotted fever
Platelet autoantibody
Drugs (acetaminophen, quinidine, sulfa drugs)
Cirrhosis Autoimmune disorders
Sepsis
Leukemia, lymphoma
Myelodysplasia
Chemo or radiation therapy
Know as thrombocytosis:Cancer (lung, gastrointestinal,breast, ovarian, lymphoma)Rheumatoid arthritis, inflammatory bowel disease, lupus
Iron deficiency anemia
Hemolytic anemia
Myeloproliferative disorder (e.g., essential thrombocythemia)
Source: American Association for Clinical Chemistry. (2014.). Complete blood count. Retrieved from http://labtestsonline.org/understanding/analytes/cbc/tab/test/ com
References
American Association for Clinical Chemistry. (2014.). Complete blood count. Retrieved from
http://labtestsonline.org/understanding/analytes/cbc/tab/test/ com American Association for Clinical Chemistry. (2014.). Platelet count. Retrieved
from http://labtestsonline.org/understanding/analytes/platelet/tab/test/Department of Medical Microbiology. (2014). A manual of clinical laboratory
diagnostic procedures. College of Public Health: UP Manila Fitch, H. (1957). An explanation of the discrepancy between direct and indirect
platelet counts. Blood: 12(7). Retrieved from http://www.bloodjournal.org/content/12/7/671?sso-checked=trueMercy Medical Center- North Iowa. (n.d.). CBC Normal Ranges. Retrieved from
http://www.mercynorthiowa.com/cbc-normal-rangesPortland Community College. (2010, December 17). 4 platelet count: Microscope
check [Video File]. Retrieved from http://www.youtube.com/watch?v=c8WuNZh1XyA
RBC Indices
• Red Blood Cells: Carry oxygen in the blood
• Measures the physical characteristics including size and the hemoglobin content of the RBCs
• Also known as Mean Corpuscular Values1. Mean Corpuscular Volume (MCV)
2. Mean Corpuscular Hemoglobin (MCH)
3. Mean Corpuscular Haemoglobin Concentration (MCHC)
Source: Epstein, J. RBC Indices. (2012). Retrieved from: http://www.healthline.com/health/rbc-indices#Overview1
Importance
• For investigation of type of anemia and for guiding therapy
1.Low RBC/ Low RBC Indices Anemia
2.High RBC/ High RBC Indices Polycythemia– Anemia: Number of RBCs or Hemoglobin is
below normal levels
• Serve as quality control procedures
1. Mean Corpuscular Volume (MCV)
Source: Curry, C.V., Mean Corpuscular Volume (MCV). (2012). Retrieved from: http://emedicine.medscape.com/article/2085770-overview#showall Red blood cell indices (indexes). (n.d.). Retrieved from: http://www.pum.edu.pl/__data/assets/file/0010/24112/unit2-medical-Red-blood-cell-indices.pdfEpstein, J. RBC Indices. (2012). Retrieved from: http://www.healthline.com/health/rbc-indices#Overview1
1. Mean Corpuscular Volume (MCV)(MCV)
– NOTE: May vary depending on the laboratory and patient’s age
Source: Curry, C.V., Mean Corpuscular Volume (MCV). (2012). Retrieved from: http://emedicine.medscape.com/article/2085770-overview#showallMoses, S. Mean Corpuscular Volume. (2009). Retrieved from: http://www.fpnotebook.com/hemeonc/lab/MnCrpsclrVlm.htm
AGE REFERENCE RANGE
Newborn 95 – 121 fL
6 mos – 2 years 70 – 86 fL
12 to 18 years
Boys 78 – 98 fL
Girls 78 – 102 fL
Adults 78 – 98 fL
2. Mean Corpuscular Hemoglobin (MCH)
Source: Epstein, J. RBC Indices. (2012). Retrieved from: http://www.healthline.com/health/rbc-indices#Overview1 Merritt, B.Y. Mean Corpuscular Hemoglobin (MCH) and Mean Corpuscular Hemoglobin Concentration (MCHC). (2014). Retrieved from: http://emedicine.medscape.com/article/2054497-overview#showall Red blood cell indices (indexes). (n.d.). Retrieved from: http://www.pum.edu.pl/__data/assets/file/0010/24112/unit2-medical-Red-blood-cell-indices.pdf
RBC Indices
• MCV and MCH have the same denominator: RBC count– MCV relates to Hemoglobin– MCH relates to Hematocrit
• The value of MCV is usually higher than MCH by approximately 3 times
Source: Department of Medical Microbiology. (2014). A manual of clinical laboratory diagnostic
procedures. College of Public Health: UP Manila
3. Mean Corpuscular Hemoglobin Concentration (MCHC)
SourceDepartment of Medical Microbiology. (2014). A manual of clinical laboratory diagnostic procedures. College of Public Health: UP Manila Epstein, J. RBC Indices. (2012). Retrieved from: http://www.healthline.com/health/rbc-indices#Overview1 Merritt, B.Y. Mean Corpuscular Hemoglobin (MCH) and Mean Corpuscular Hemoglobin Concentration (MCHC). (2014). Retrieved from: http://emedicine.medscape.com/article/2054497-overview#showall Red blood cell indices (indexes). (n.d.). Retrieved from: http://www.pum.edu.pl/__data/assets/file/0010/24112/unit2-medical-Red-blood-cell- indices.pdf
Reference Range Female: Male:
MCHC 30-34 gHb/100ml 30-34 gHb/100mL
NOTE: May vary depending on the laboratory and patient’s age
RBC Indices
Source: Blood cell indices. (n.d.) Retrieved from: http://www.medicine.mcgill.ca/physio/vlab/bloodlab/mcv-mchc_n.htm
Causes of Blood Abnormalities
1. MCH and MCHC mirror MCV– Larger RBC means higher amount/concentration
of Hbg2. ↑ MCHC
– severe burns or hereditary spherocytosis3. Normocytic/normochromic anemia
a) Sudden and significant blood lossb) Prosthetic heart valvec) Tumord) Chronic disease e) Aplastic anemia
Source: Epstein, J. RBC Indices. (2012). Retrieved from: http://www.healthline.com/health/rbc-indices#Overview1
Causes of Blood Abnormalities
6. Macrocytic/normochromic anemia:a) B-12 and/or folate deficiencyb) Chemotherapy
7. Microcytic/normochromic anemia– Kidney Failure
8. Microcytic/hypochromic anemiaa) Iron deficiency , b) Lead poisoningc) Thalassemia
9. Polycythemia vera– Disorder of the bone marrow due to genetic mutation
of JAK2 Source: Epstein, J. RBC Indices. (2012). Retrieved from: http://www.healthline.com/health/rbc-indices#Overview1
References
Blood cell indices. (n.d.) Retrieved from: http://www.medicine.mcgill.ca/physio/vlab/bloodlab/mcv-mchc_n.htm
Curry, C.V., Mean Corpuscular Volume (MCV). (2012). Retrieved from: http://emedicine.medscape.com/article/2085770-overview#showall
Department of Medical Microbiology. (2014). A manual of clinical laboratory diagnostic procedures. College of Public Health: UP Manila
Epstein, J. RBC Indices. (2012). Retrieved from: http://www.healthline.com/health/rbc-indices#Overview1
Merritt, B.Y. Mean Corapuscular Hemoglobin (MCH) and Mean Corpuscular Hemoglobin Concentration (MCHC). (2014). Retrieved from: http://emedicine.medscape.com/article/2054497-overview#showall
Moses, S. Mean Corpuscular Volume. (2009). Retrieved from: http://www.fpnotebook.com/hemeonc/lab/MnCrpsclrVlm.htm
Red blood cell indices (indexes). (n.d.). Retrieved from: http://www.pum.edu.pl/__data/assets/file/0010/24112/unit2-medical-Red-blood-cell-indices.pdf