EXERCISE 9: AUTOMATED HEMOGLOBIN TESTING
Contents
1. PROCEDURES Performing the Blank Reading Perform a blank reading whenever you have removed the cuvette holder i e cleaning replacing with a new cuvette holder Open the cuvette holder completely The text in the display reads Add Cuvette Close the cuvette holder without inserting a cuvette In this case the photometer does not start testing After approx 2 3 seconds the display will read Open Holder If the instrument DOES NOT show Open Holder there is possibly a malfunction Please consult Troubleshooting 5 Record on the Recording Results Form Pee Testing the Control Cuvette 1 Open the cuvette holder completely and wait until the photometer displays Add Cuvette 2 Take the control cuvette out of the storage box and place it in the cuvette holder 3 Close the cuvette holder completely The photometer tests the control cuvette and shows the result after a few seconds 4 Compare the result with that stated on the storage box label The result must lie within the stated range Record the control result 5 If the Hgb result falls outside the range of the control cuvette there is evidently a problem Please consult Section 7 Troubleshooting in this regard 6 Record on the Recording Results Form If the original control cuvette is lost or damaged the photometer must be checked for proper calibration and a new control cuvette must be assayed The photometer will nee2. Cleaning the Optical Unit 1 il clad SOPENA 11 0 The optical unit is situated inside the photometer and has no direct contact with the cuvette therefore no routine cleaning is needed Cleaning the optical unit can become necessary if the measured maximum light intensity of the photometric light source no longer achieves the appropriate level required for testing For cleaning the optical unit the use of a special HemoPoint H2 optics cleaner is recommended Pull the cuvette holder out while the photometer is in the ready mode Remove the cleaner from the foil pouch and insert it narrow tip first carefully into the opening of the cuvette holder until you feel a smooth resistance THE CLEANER MUST BE USED WITHIN 10 MINUTES OF OPENING Slowly push the cleaner deeper into the opening until it stops Wipe the optical system several times by slowly moving the cleaner in and out at least 5 times Remove the cleaner from the photometer If the used cleaner is dirty repeat the procedure with a new cleaner Wait at least 15 minutes after the optical system has been cleaned to reinsert the cuvette holder into the photometer Dispose of all used cleaners as potentially infectious waste Exercise 7 Automated Hemoglobin Testing rev 12 7 2013 EXERCISE 9 AUTOMATED HEMOGLOBIN TESTING Phlebotomist Date Patient Name Points 34 possible INSTRUCTIONS Place a check in the appropriate column of the form P indicates
3. Normal 26 High Patient Name Results in gm dL Points 2T 28 Based on the results of the control solutions can this machine be used circle one YES NO 29 Are the patient results circle one NORMAL ABNORMAL Exercise 7 Automated Hemoglobin Testing rev 12 7 2013 EXERCISE 9 AUTOMATED HEMOGLOBIN TESTING STUDY QUESTIONS Name Date Points 26 1 Define the following terms 2 5 points a hemoglobin b anemia c polycythemia d sulfhemoglobin e carboxyhemoglobin De State two possible causes of low and high hemoglobin values 2 points a Low i il b High 1 il 3 State the principle of the HemoPoint H2 hemoglobin procedure 1 point 4 State three criteria which must be considered when setting up the HemoPoint photometer 3 points a b c 5 List 3 sample types which may be used for testing 1 5 points a b Exercise 7 Automated Hemoglobin Testing rev 12 7 2013 10 11 12 13 State the storage temperature requirements and time limits for use of blood samples 2 points a Storage b Time limit State when a blank reading of a microcuvette is required 1 point State the frequency of testing the control cuvette 1 point Describe the proper handling and storage of cuvettes including temperature amount of time they can be used after opening and the information to be placed on the container the first time ope
4. 200 DY el 11 12 17 18 19 20 21 22 EXERCISE 7 AUTOMATED HEMOGLOBIN TESTING Textbook Skills 34 points Objectives Define the following hemoglobin anemia polycythemia sulfhemoglobin carboxyhemoglobin State two possible causes of low and high hemoglobin values State the principle of the HemoPoint H2 hemoglobin procedure State six criteria which must be considered when setting up the HemoPoint photometer List 3 sample types and 3 anticoagulants which may be used for testing State the storage requirements and time limits for use of venous and arterial samples Describe the testing which must be performed for monitoring quality assurance in hemoglobin testing State when a blank reading of a microcuvette is required State the frequency of testing the control cuvette and the importance of only using the control cuvette designated for each instrument State the proper handling and storage of cuvettes including temperature amount of time they can be used after opening and the information to be placed on the container the first time opened List 8 limitations of the hemoglobin test procedure State what must be done if hemoglobin values are equal or greater than 23 5 g dL 13 14 15 16 State the normal values for hemoglobin levels for adult females adult males children and infants Perform and record the blank reading and testing the control cuvette Perform and record the calibr
5. with which it is delivered i e the Hgb value stated on the storage box label is only valid for that one photometer and could lead to completely different results on another photometer If you have several HemoPoint H2 photometers keep track of each control cuvette for each photometer When not being used keep the control cuvette in the original storage box It is optimally protected there against breakage and contamination External Quality Control Controls when assayed as actual specimens help a laboratory evaluate whether a given procedure is performing with acceptable accuracy and precision Laboratory Practices recommend the daily use of external controls to assure that the microcuvettes and the photometer are performing correctly For this purpose we recommend the use of Stanbio HemoPoint H2 Hemoglobin Controls Three 3 different control preparations are provided in the set These are prepared from unfixed stabilized human erythrocytes containing normal low and high levels of hemoglobin With the help of these three 3 different concentrations you can easily assess the precision of the system There are a variety of hemoglobin controls on the market today Stanbio only recommends the use of the HemoPoint H2 Hemoglobin Controls since we cannot guarantee the compatibility of other brands of controls on the HemoPoint H2 system Thoroughly mix the controls before use see the control package insert for directions Exer
6. ation and control solution results for the test Evaluate the calibration and normal control solution using predetermined criteria to determine if the test can be performed on the patient Perform an effective capillary puncture Perform a quantitative measurement of hemoglobin on whole blood using the HemoPoint H2 instrument Evaluate the patient results and determine whether they fall within the normal range State the time limit for testing the sample in the microcuvette Record the results using proper units After use perform maintenance on the housing unit touch screen cuvette holder and optical unit 23 24 State the length of time before the equipment can be used after cleaning the optical unit State the time limit for using the optical cleaner after removal from the foil pouch Discussion Hemoglobin is the oxygen carrying pigment and main component of red blood cells Low hemoglobin levels may indicate anemia recent hemorrhage or fluid retention Elevated hemoglobin levels may indicate hemoconcentration from polycythemia or dehydration The HemoPoint H2 system is intended for the quantitative determination of hemoglobin Hgb in whole blood of adults infants and children in a professional point of care setting It consists of a dedicated photometer and individual single use microcuvettes filled with reagents Using a single drop of blood Stanbio s HemoPoint H2 offers accurate results for both hemoglobin and hemat
7. cise 7 Automated Hemoglobin Testing rev 12 7 2013 Compare the values obtained with the controls against the stated reference values If the Hgb result falls outside the range of the controls there is evidently a problem Please consult Section 7 Troubleshooting in this regard Storage of Cuvettes The microcuvettes consist of a clear body with a cavity which takes up approximately 10uL of blood which combines with the dry reagent chemistry The optical distance between the cuvette walls is fixed and permits photometric determination of hemoglobin in undiluted blood samples 1 Store the cuvettes in the original container at room temperature 15 30C 59 86F DO NOT refrigerate Cuvettes are stable for 3 months after opening the vial when stored at room temperature Write the date the vial was opened and your initials on the container label in the space provided Only remove ONE microcuvette at a time from the container and then immediately close the lid It is crucial not to touch the optical eye of the cuvette with fingers dirty or sharp objects hl all ad Limitations of the Procedure 1 The microcuvette sample can be measured immediately or within 10 minutes at the latest After 10 minutes false results may be obtained 2 Air bubbles in the optical eye caused by inadequate filling of the microcuvette cavity may cause false results Discard the microcuvette and take another sample using a new microcuvette 3 Ensure that you do no
8. d to be sent back to Stanbio Laboratory for a new control cuvette assignment The calibration of the photometer and the re assay of a new control cuvette is a critical and extensive process The cost for this is the responsibility of the owner so please take care of the control cuvette Performing External Quality Control Commercial liquid controls that are recommended by Stanbio must be used on each day of testing to assure proper functioning of the entire system Follow the manufacturer s procedure for storage and handling of the control material 1 Allow the vials to come to room temperature 59 86 F for 20 minutes 2 Mix thoroughly but gently by inverting the vials and repeatedly rolling them between the palms until all cellular components are completely suspended Do not shake the vial Check the bottom of the vial to ensure that all cells are completely suspended and not settled in the bottom of the vial A mechanical mixer is not recommended for the procedure but can be used to maintain the cell suspension 3 The analyzer should be in the ready mode prior to filling the cuvette 4 Remove the cap from the vial Dispense a drop of control onto a clean NON ABSORBENT surface such as plastic film 5 Follow Steps 14 18 of the testing procedure below Exercise 7 Automated Hemoglobin Testing rev 12 7 2013 2 ve Values should be read within 2 minutes after filling the sample into the microcuvette While waiting for the re
9. gerator and bring it to room temperature 2 Mix the sample tube well i e by a mechanical rotator or hand inversion at least 10 times Exercise 7 Automated Hemoglobin Testing rev 12 7 2013 Quality Control The HemoPoint H2 AutoCheck performs an internal check of the photometer s optic system every time the cuvette holder is opened The following checks are performed each day of use 1 The control cuvette supplied with the photometer can be used for a simple check of the photometer s calibration 2 Use of external controls to assure that the microcuvettes and the photometer are performing correctly Blank reading All the optical and mechanical components involved with the measuring cycle are carefully manufactured with extremely small tolerances This is essential for the proper function of the photometer Always carry out a blank reading whenever you have removed the cuvette holder i e cleaning replacing with a new cuvette holder Control Cuvette The control cuvette supplied with the photometer can be used for a simple check of the photometer s calibration The testing quality of the photometer should be checked once daily using this control cuvette The Hemoglobin value and the permitted deviation of the control cuvette are stated on the storage box label All photometers have a specific calibration due to the tolerances in the mechanical and electronic components The control cuvette is only calibrated for the instrument
10. h Exercise 7 Automated Hemoglobin Testing rev 12 7 2013 Maintenance Cleaning and disinfection of the instrument after use Cleaning the Housing and Touch screen 1 2 Disconnect the power adaptor from the electrical connection before proceeding Cleaning the housing and touch screen is best accomplished with a lint free cloth lightly dampened with clean water For more stubborn soiling a mild soap solution may be used For disinfection standard solutions can be used for surface disinfection provided they do not contain alcohol or other solvent Cleaning the Cuvette Holder 1 2 The cuvette holder can be removed from the instrument for cleaning Proceed as follows a Disconnect the power adaptor from the electrical connection before proceeding b Open the cuvette holder until you feel a resistance and the holder will not extend further c Press down the silver pin on the left hand side bottom of the cuvette holder with a ball point pen and draw the cuvette holder forward at the same time The cuvette holder can now be cleaned with a mild soap solution For disinfection standard solvent free preparations can be used DO NOT use any cleaning agent for cleaning the cuvette holder that could leave scratches on its surface Wait until the cuvette holder is completely dry before returning it to the machine To replace the cuvette holder gently push it in the correct position into the opening in the housing until it engages
11. ned 3 points a Temperature b Time of use after opening c Information on container List 4 limitations of the hemoglobin test procedure 4 points a b c d What must be done if hemoglobin values are equal or greater than 23 5 g dL 1 point State the normal values for hemoglobin levels for adult females adult males children and infants 2 points a Adult female b Adult male Children d Infants State the following time limits a Testing the sample once it is collected in the microcuvette 0 5 point b Vials of control to come to room temperature 0 5 point G Optical cleaner be used after removal from the foil pouch 0 5 point d Instrument be used after cleaning the optical unit 0 5 point 10
12. ocrit calculated tests in less than a minute The easy to read touch screen displays the current time battery status and allows for easy operation Exercise 7 Automated Hemoglobin Testing rev 12 7 2013 Using the microcuvette a small amount of arterial venous or capillary blood is taken up by capillary action The filled cuvette is inserted into the HemoPoint H2 photometer the color produced by the chemical reaction in the cuvette is measured and the Hgb value is displayed The microcuvettes are intended to be used only once and must be disposed after use as potentially infectious waste in accordance with the current regulations applicable to your establishment The HemoPoint H2 system is designed for use in medical practices and in clinical laboratories to assist in medical diagnostic investigations In addition it can be used in emergency and intensive care units and in medical facilities such as blood banks This test system has been approved for use in laboratories with Waived status as defined in 452 CFR 493 15 c a Principle of the Test Procedure In the HemoPoint H2 system the use of microcuvettes with short light pathways makes it possible to analyze undiluted blood The filled microcuvette is inserted into the HemoPoint H2 photometer the color produced by the chemical reaction in the cuvette is measured and the hemoglobin level is calculated and displayed For this purpose light is directed through the blood sam
13. ple and the absorption is measured From the amount of light absorbed by the sample the concentration of the hemoglobin in the cuvette can be calculated Setting up the Photometer Select a suitable place for setting up the photometer according to the following criteria 1 Avoid direct sunlight 2 Avoid strong electromagnetic fields 3 Avoid direct influence from ionizing radiation 4 Avoid rapid temperature fluctuations keep away from heaters open windows ventilators fans or air conditioning etc Avoid wet areas i e wash basins Allow the photometer to reach room temperature Changing from a cold to a warm environment i e after storage or transport condensation can form both on the inside and the outside of the photometer Wait at least 1 hour before you connect the photometer to a power supply NM Place the photometer on a level counter adjacent to a power socket Make sure there is enough room for the cuvette holder to be freely accessible Sample Collection and Preparation The HemoPoint H2 Photometer can be used with capillary venous or arterial blood Use EDTA heparin or heparin fluoride as anticoagulants preferably in solid form to avoid dilutional effects Venous and arterial blood samples may be used if the blood collected is not more than 24 hours old and the samples have been stored refrigerated 35 46F 2 8 C Prepare stored samples for measurement as follows 1 Remove sample tube from the refri
14. successful performance NP indicates not performed P NP 1 Performs blank reading in accordance with procedure 2 Test control cuvette in accordance with procedure States the proper protocol for the performance of quality control procedures Role playing Introduce yourself Role playing Identify the patient Role playing Explain the procedure Disinfects hands and put on gloves OO VN a Selects prepares and organizes equipment 9 Selects puncture site massages and warms site as necessary 10 Cleanses site allows site to dry completely 11 Performs an effective capillary puncture 12 Appropriately squeezes finger to form first drop of blood 13 Wipes away first drop of blood 14 Applies blood to microcuvette without bubbles 15 If necessary wipes off excess blood 16 Places microcuvette in loading tray and inserts into the machine 17 Reads and records results 18 Provides appropriate post puncture care to the patient 19 Disposes of all materials and supplies in the appropriate containers 20 Inspects puncture site applies bandage if necessary 21 Removes gloves washes hands Acceptable NA Yes No 22 Blank reading performed 23 Testing control cuvette Control Solutions Value Obtained gm dL Manufacturer Values 24 Low 25
15. sults clean the threads of the vial and threads of the cap with a cotton or polyester gauze Snugly recap the vial immediately After testing return the vial to its proper storage Record the results on a quality control section of the Recording Results Form Expected ranges for each level are on the package insert which comes with a specific lot number of controls If the results do not fall within the established range repeat the control test If results still are outside of established ranges consult the User s Manual or contact HemoPoint technical support at 800 531 5535 TESTING CANNOT BE PERFORMED IF THE RESULTS OF THE CONTROLS ARE OUT OF RANGE Procedure Performing the Test MAW PS YM ES 10 11 12 13 14 15 16 17 18 19 Make sure the Photometer is ready for use Test control cuvette if required blank reading and quality control Identify yourself and your patient Disinfect your hands and put on gloves Explain the procedure to your patient Make sure that your patient is sitting comfortably Remove ONE cuvette from the container and IMMEDIATELY close the lid NOTE If this is the first cuvette removed WRITE THE DATE and YOUR INITIALS on the container in the space provided Take care NOT to touch the optical eye of the cuvette with your fingers There should be good blood circulation in the hand from which you wish to take blood e g it should be warm and relaxed Lightly massage
16. t hold the microcuvette at its filling end this may contaminate the optical eye preventing an accurate reading 4 To avoid contamination of the cuvette holder remove surplus blood from the outside of the microcuvette 5 Always place the cuvette right side up in the holder Placing the cuvette upside down can lead to erroneous results 6 All results above 23 5 g dL or equivalent must be confirmed by laboratory methods 7 Sulfhemoglobin a form of hemoglobin found in the blood in trace amounts that contains an irreversibly bound sulfur molecule that prevents normal oxygen binding Cannot be measured by this method 8 Carboxyhemoglobin a stable complex of carbon monoxide and hemoglobin that forms in red blood cells when carbon monoxide is inhaled and hinders delivery of oxygen to the body formed in carbon monoxide poisoning or tobacco smoking and turbidity due to leukocytosis or hyperlipemia DO NOT interfere Expected Values Normal range Reference Values Adult Males 13 0 18 0 g dL Adult Females 11 0 16 0 g dL Children 2 yrs to teenage 11 0 16 0 g dL Infants post natal 10 14 g dL The highest hemoglobin concentrations are usually measured in neonates Due to the wide range of conditions dietary geographical smoking exercise recumbency etc which affect reference values it is recommended that each laboratory establish its own expected ranges Exercise 7 Automated Hemoglobin Testing rev 12 7 2013
17. the fingers to stimulate circulation Disinfect the puncture site and allow to dry Take out a microcuvette from the container and close the lid immediately Press lightly on the fingertip and puncture with a suitable sampling device on the side of the finger WIPE AWAY the first drop of blood then if necessary press gently once again to get second drop of blood which is large enough to fill the microcuvette completely AVOID milking the finger or using excess pressure Hold the tip of the microcuvette tip in the middle of the drop of blood and let the cavity fill in one step Visually inspect the cuvette for air bubbles in the optical eye In case of air bubbles in the optical eye of the microcuvette discard and take another sample using a new microcuvette In order to avoid contamination of the cuvette holder remove surplus blood from the outside of the microcuvette by gently wiping with a biowipe NOTE The microcuvette sample prepared this way can now be measured immediately or within 10 minutes at the latest Immediately insert the cuvette into the cuvette holder The result will be displayed within 10 seconds to 3 minutes and will remain on the display as long as the cuvette holder is in the measuring position Record the results of the hemoglobin and hematocrit USING APPROPRIATE UNITS After testing is complete gently pull the cuvette holder out to the loading position and properly dispose of the cuvette in biohazardous tras
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