hema project

Hematologic ExaminationsPrepared by: Group 6 (Chito, Chua, Co, Cortez)

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


• Measured using a hemoglobinometer

• Procedure:

Source: www.medicine.mcgill.ca/physio/vlab/bloodlab/hemoglobin_n.htm


1.Drop of blood is placed on one side of the chamber glass


2. Blood is stirred with a hemolysis applicator, with saponin


3. Cover glass is placed on top of chamber glass.4. Both are slid onto metal clip.


5. Blood chamber is inserted into a slot in the hemoglobinometer.


• 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


• 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


• 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


• 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


• 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: 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


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)



• Decreased Hematocrit– Hemorrhage– Chronic kidney disease– Inadequate iron uptake– Iron deficiency due to blood loss during menstruation– Anemia– Malnutrition– Overhydration



• A hematocrit of less than 15% can result in cardiac failure

• A hematocrit of over 60% may result in spontaneous blood clotting


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


Platelet Count

2 Methods of Platelet Counting

1.Direct Platelet Count

2.Indirect Platelet Count


Direct Platelet Count


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


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



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


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


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

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