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1 LESSON 6-3 Laboratory Reagent Preparation and Calculations Worksheet LESSON 6-4 Chemistry Instrumentation in the Physician Office Laboratory Worksheet LESSON 6-6 Measuring Blood Glucose Worksheet LESSON 6-6 Measuring Blood Glucose Worksheet LESSON 6-7 Measuring Electrolytes Student Performance Guide LESSON 6-8 Measuring Blood Cholesterol Student Performance Guide

2 Worksheet LESSON 6-3 Laboratory Reagent Preparation and Calculations Name Date 1. A procedure calls for 200 ml of a 2% glucose solution. A 50% solution is available. How much of the 50% solution is needed? How would the solution be prepared? 2. A 1% solution of hydrochloric acid is required for a procedure. A 5% solution is available. How much of the 5% solution will be required to make 500 ml of a 1% solution? 3. A procedure calls for acetic acid and water, with the proportions being two parts acetic acid to three parts water. One hundred milliliters are needed. How much acetic acid and water are required? 4. One liter of 70% alcohol is needed. How much 95% alcohol is required to make the 70% solution?

3 5. Two hundred milliliters of 2% acetic acid are needed. How many ml of 5% acetic acid are required to make the 2% solution? 6. How would one liter of a 10% solution of chlorine bleach be prepared? 7. Give the instructions for preparing one liter of a 0.5N solution of Mg (OH) 2 (FW = 58; valence = 2). 8. How could 250 ml of a 4% solution of hydrochloric acid (HCl) be prepared from a 10% solution of HCl? 9. Give the instructions for preparing serial dilutions of a serum sample using 0.5 ml serum and 0.5 ml saline in each of five numbered tubes.

4 10. A 1-to-25 dilution of blood is required for a procedure. How is it prepared? 11. How could a 1:10 dilution of serum be prepared? 12. What is the dilution when 0.5 ml is diluted to a total of 100 ml? 13. If 0.5 ml of serum is added to 4.5 ml of saline, what is the dilution? 14. If 0.5 ml of blood is added to 9.5 ml of saline, what is the dilution?

5 Worksheet LESSON 6-4 Chemistry Instrumentation in the Physician Office Laboratory Name Date Part A. Answer the following questions: 1. What types of technologies are used in most clinical chemistry analyzers? 2. How do the requirements for a POL instrument differ from those for an instrument in a large hospital laboratory? 3. What is the advantage of an instrument that can analyze a whole blood sample instead of only serum or plasma? 4. List five procedural notes to follow when an instrument is being used for analysis. 5. Why would the information in a maintenance logbook be important when a problem arises with an instrument? Part B. Read the manufacturer s operatoring manual (or laboratory procedure manual) for an instrument available in the laboratory and answer the following questions: 7. What is the name of the instrument? 8. Does this instrument use solid-phase, discrete, or ion-selective electrode technology? 9. If the instrument does not fit into any of these categories, what type of technology is involved? 10. List two test procedures that can be performed using this instrument. 11. How does the worker tell the instrument which test procedure is to be performed? 12. How are control solutions used with this instrument?

6 13. Has a Levey-Jennings chart been constructed for the procedures run on this instrument? Are values plotted daily? 14. Is plasma, serum, or whole blood used for the analyses performed on this instrument? 15. Are maintenance and repair records kept in a logbook? 16. List in order the steps for performing a glucose (or other) analysis using this instrument.

7 Worksheet LESSON 6-6 Measuring Blood Glucose Name Date INSTRUCTIONS 1. Practice the procedure for measuring glucose using a glucose analyzer and following the step-by-step procedure. 2. Demonstrate your understanding of this lesson by: a. Completing a written examination successfully, and b. Using the glucose analyzer to measure glucose satisfactorily for the instructor. All steps must be completed as listed on the instructor s Performance Check Sheet. MATERIALS AND EQUIPMENT gloves glucose meter such as HemoCue, Accu-Chek, or Glucometer test strips, cuvettes, or cartridges for glucose meter control solutions for glucose meter soft laboratory tissue capillary puncture materials hand disinfectant surface disinfectant (10% chlorine bleach solution) biohazard container worksheet 70% alcohol and cotton balls, or alcohol swabs micropipetter µl, (optional depending on analyzer used) puncture-proof sharps container Note: The following is a general procedure for using a glucose analyzer or glucose meter. Consult the procedure manual for the instrument used and package inserts for specific instructions. PROCEDURE Record in the comment section any problems encountered while practicing the procedure (or have a fellow student or the instructor evaluate your performance). S = Satisfactory U = Unsatisfactory You must: S U Comments 1. Review operating instructions for the glucose analyzer 2. Wash hands and put on gloves 3. Assemble materials and supplies 4. Record instrument name, control ranges, control lot #, and test strip lot # on the worksheet 5. Measure blood glucose using the Accu-Chek Advantage by following steps 5a 5h (For the HemoCue, skip to step 6; for other meter, go to step 7): a. Perform quality control check: (1) Turn on the meter. Check that code on display matches code on test strip vial

8 You must: S U Comments (2) Insert Chek Strip into monitor (3) Note that OK appears if monitor is functioning properly (4) Remove Chek Strip and return it to storage (5) Insert a test strip into the meter when the test strip symbol flashes (within thirty seconds). Wait until the display indicates it is ready for the specimen (display will read L1 ) (6) Apply one drop of Level 1 control to the target area on the test strip (7) Record the displayed value on the quality control log. Verify that control solution is within acceptable range (Display will read OK ) (8) Discard used test strip (9) When L2 appears on display, insert a new strip into meter. Repeat steps 5a6 5a8 using Level 2 control b. When meter indicates it is ready, insert a new test strip c. Perform capillary puncture on patient d. Hold finger over test strip and apply one large hanging drop of blood to target area on strip, being certain that entire area is covered with blood e. Read the patient s glucose value from the meter and record f. Discard the test strip in biohazard container g. Turn off the meter and return it to storage 6. Perform a blood glucose measurement using the HemoCue following steps 6a-6k: a. Turn on the meter b. Insert the control cuvette into the meter. Check to see that instrument is calibrated. c. Fill a cuvette with a glucose control solution following the manufacturer s directions d. Place the cuvette in the carrier and gently push it into the HemoCue. Read the value displayed and record on quality control log. Verify that control solution is within acceptable range e. Discard cuvette f. Obtain a new cuvette g. Perform capillary puncture on patient h. Fill the cuvette from the capillary puncture. Do not allow air bubbles into the cuvette i. Place the cuvette in the carrier and gently push it into the HemoCue j. Read the patient s glucose value from the display and record k. Discard the cuvette in a biohazard container l. Turn off the meter, wipe it with disinfectant, if necessary, and return it to storage 7. Perform a blood glucose measurement using a glucose meter: a. Perform quality control check following manufacturer s instructions b. Perform capillary puncture on patient

9 You must: S U Comments c. Apply blood to test strip or cartridge and insert into meter according to manufacturer s instructions d. Read the patient s glucose value from the meter and record e. Discard the test strip in biohazard container f. Turn off the meter and return it to storage 8. Discard used capillary puncture materials in appropriate biohazard containers 9. Disinfect work area with surface disinfectant 10. Remove and discard gloves in biohazard container and wash hands with hand disinfectant. Evaluator Comments: Evaluator Date

10 Worksheet LESSON 6-6 Measuring Blood Glucose Name Date 1. Instrument Name: 2. Control test: Record lot numbers and acceptable ranges of glucose controls and reagent test unit. Record results of control test(s). If test(s) is within range, mark acceptable (A); if not, mark unacceptable (U). Test Unit Lot # Glucose Controls Lot No. Acceptable Range Control Results A U Normal Low High Action taken if control test is unacceptable: 3. Patient test: Record the results below and compare with the normal range given. Patient I.D. Test Result Reference Range (whole blood) mg/dl

11 Student Performance Guide LESSON 6-7 Measuring Electrolytes Name Date INSTRUCTIONS 1. Practice measuring electrolytes using an analyzer and following the step-by-step procedure. 2. Demonstrate your understanding of this lesson by: a. Completing a written examination successfully, and b. Performing the procedure for measuring electrolytes as directed by the instructor. All steps must be completed as listed on the instructor s Performance Check Sheet. Note: The following is a general procedure for measuring electrolytes. The manufacturers instructions for the specific methods or instruments being used must be consulted and carefully followed. MATERIALS AND EQUIPMENT gloves serum or plasma specimens blood-collection equipment for: syringe venipuncture vacuum-tube venipuncture capillary puncture i-stat analyzer or other small analyzer capable of measuring electrolytes, including test cartridges and necessary controls quality control charts surface disinfectant biohazard container biohazard sharps container PROCEDURE Record in the comment section any problems encountered while practicing the procedure (or have a fellow student or the instructor evaluate your performance). S = Satisfactory U = Unsatisfactory You must: S U Comments 1. Assemble equipment and materials 2. Wash hands and put on gloves 3. Turn on instrument 4. Perform quality control checks as directed by manufacturer and record results on quality control chart 5. Obtain test cartridge or cassette 6. Perform capillary puncture (or use previously obtained blood specimen)

12 You must: S U Comments 7. Apply blood specimen to test cartridge 8. Insert cartridge into instrument 9. Read and record test results 10. Discard test cartridge in biohazard container 11. Turn off instrument and store according to manufacturer s directions 12. Discard all capillary puncture materials appropriately 13. Clean work surface with surface disinfectant 14. Remove and discard gloves and wash hands with hand disinfectant Evaluator Comments: Evaluator Date

13 Student Performance Guide LESSON 6-8 Measuring Blood Cholesterol Name Date INSTRUCTIONS 1. Practice the procedure for measuring blood cholesterol following the step-by-step procedure. 2. Demonstrate your understanding of this lesson by: a. Completing a written examination successfully, and b. Performing the procedure for measuring blood cholesterol satisfactorily for the instructor. All steps must be completed as listed on the instructor s Performance Check Sheet. Note: The following procedure is a general procedure for measuring cholesterol using Sigma Diagnostics Procedure No Consult the manufacturer s instructions for the specific method being used. MATERIALS AND EQUIPMENT gloves blood-collecting equipment hand disinfectant spectrophotometer commercial kit for manual determination of cholesterol (or use an instrument, following all of the manufacturer s instructions) serum controls and cholesterol standards marking pencil test tubes (13 x 100 mm) test tube rack cuvettes for spectrophotometer laboratory tissue Parafilm calculator (optional) pipetter with disposable tips water bath at 30 C or 37 C (optional) surface disinfectant (10% chlorine bleach solution) biohazard container instrument for performing cholesterol determination, including test materials, controls, and standards Levey-Jennings chart biohazard sharps container PROCEDURE Record in the comment section any problems encountered while practicing the procedure (or have a fellow student or the instructor evaluate your performance). S = Satisfactory U = Unsatisfactory You must: S U Comments 1. Assemble equipment and materials 2. Wash hands and put on gloves 3. Obtain blood sample from the patient, either by fingerstick or venipuncture, depending on the type of sample required

14 You must: S U Comments 4. Perform either method a or b: a. Manual method (Sigma Diagnostics Procedure No. 352) (1) Prepare Cholesterol Reagent according to instructions (2) Turn on spectrophotometer and set wavelength to 500 nm (3) Set the absorbance reading to zero using water as the blank (reference) (4) If using spectrophotometer with temperature-controlled cuvette compartment, set temperature to 37 C, or the assay may be performed at ambient (room) temperature, or using a water bath at 30 or 37 C (5) Label cuvettes or test tubes for Blank, Calibrator, Control, and Sample (6) Warm reagent to the temperature being used for the assay (7) Pipet 1.0 ml of reagent into each of the prepared tubes (8) Add 0.01 ml (10 µl) deionized water to the Blank (9) Add 10 µl of Calibrator, Control, and Sample to the appropriately labeled tubes. Cover tubes with Parafilm and mix tubes by gentle inversion (10) Incubate tubes or cuvettes for five minutes at 37 C or ten minutes at C (11) Read and record absorbances of all tubes at 500 nm. Complete the readings within thirty minutes after the end of incubation time (12) Calculate total cholesterol in sample and control, using the absorbance formula (below). Record the results Serum A TEST A BLANK cholesterol = x Calibrator A (mg/dl) CALIBRATOR A BLANK (mg/dl) b. Automated method (general) (1) Turn on the instrument (2) Wash hands and put on gloves (3) Prepare any control or calibrator reagents (4) Run appropriate controls or calibrator samples (5) Obtain correct patient specimen (6) Choose appropriate test strip or pack for cholesterol determination (7) Add patient sample to test system (8) Insert strip or pack in instrument (9) Ensure that correct test is being performed (10) Wait for results to be displayed or printed out (11) Record results 5. For each method, add the control values to a Levey-Jennings chart 6. If the method is in control, report patient values 7. Disinfect and clean reusable equipment

15 You must: S U Comments 8. Dispose of contaminated materials in biohazard container or sharps container 9. Wipe work area with surface disinfectant 10. Turn instrument OFF (or leave as manual instructs) 11. Remove gloves and discard in biohazard container 12. Wash hands with hand disinfectant Evaluator Comments: Evaluator Date