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Dimension® EXL™ with LM Integrated Chemistry System HM & LOCI® Online Training

Locate system components on the HM and LOCI® areas. Perform routine maintenance tasks. Determine need for replacement of LOCI® parts. This clinical laboratory training qualifies for continuing education units (CEU).

Describe the HM reaction and processing steps Describe the homogeneous processing using LOCI® technology Identify consumables used by the HM and LOCI® systems Identify key components and their functions for both heterogeneous (HM) and homogeneous (LOCI®) testing Welcome to Assay Measurement  training course for the HM and LOCI®  technologies on the Dimension®  EXL™ with LM integrated chemistry system. After completing this course, you will be able to: Select Next to continue. The Heterogeneous Immunoassay Module (HM) is used to process chromium-based immunoassays for the measurement of specialty analytes of low molecular weight. The immunoassay is prepared in the HM area, and the final results are read photometrically after transferring reactants from reaction vessels to cuvettes.   Other HM module features include:     Disposable reaction vessels to set up the immunoassay  Automatic loading of reaction vessels into the incubate wheel via the vessel holder and track system Chromium-based antibody with high sensitivity  Increased processing throughput by performing immunoassay steps concurrently rather than sequentially HM Features Learn about the most important feature of the HM module: the heterogeneous immunoassay. Slide NumberText BlocksCalloutsAudio ScriptImage File1 In this section,  you will learn how HM methods are processed. Select Next to continue.Note: If audio does not automatically start, select the play arrow in the top left to begin.In this section, you will learn how HM methods are processed.2 All Flex® reagent cartridges used for HM testing have two components in common: Chromium dioxide coated with capture antibody Conjugate/tag antibody reagent Select Next to continue.All Flex® reagent cartridges used for HM testing have two common components: Chromium dioxide coated with capture antibody, and Conjugate/ tag antibody reagent. As reagent components, these antibodies are used to detect or measure specific antigens in a sample. Antigens are the analytes that will be measured during the heterogeneous immunoassay. 3 The type of immunoassay formats used for the HM methods depends on the particular analyte being tested. Note: Refer to the Flex® reagent cartridge Instructions for Use (IFU) sheet for details about each HM test method. Select Next to continue. Select each number to review the corresponding text.CalloutsTag AbTag Ab CrO2/Capture AbCrO2/Capture Ab CPRGCPRG CRPG DiluentCrO2 DiluentThe type of immunoassay formats used for the HM methods depends on the particular analyte being tested.4 R2 reagent probe adds chromium-based antibody to the reaction vessel on the incubate wheel. Select Next to continue. Select the number to review the corresponding text.CalloutsCrO2 added by R2 from Flex reagent cartridgeIn this example, let's process an HM test request for an MMB level. To start the assay, the R2 probe adds chromium dioxide from the Flex reagent cartridge to the reaction vessel on the incubate wheel. Chromium dioxide is coated with capture antibody specific for the analyte being tested.5 Photometric sample probe adds sample to the reaction vessel. Select Next to continue. Select the number to review the corresponding text.CalloutsSample containing analyte added from cup or tube. The photometric sample probe adds patient or QC sample from the sample wheel into the same reaction vessel on the incubate wheel. The reaction vessel now contains capture Ab + sample. 6 R2 probe adds conjugate/tag antibody to the reaction vessel. Select Next to continue. Select the number to review the corresponding text.CalloutsConjugate added by R2 from Flex® reagent cartridgeThe R2 probe adds conjugate reagent from the Flex® cartridge to the reaction vessel. The conjugate/tag antibody is specific for the analyte being tested. The reaction vessel now contains capture Ab + sample + tag Ab. 7 Vessel reactants incubate at 37 degrees to form immunosandwiches. Select Next to continue. Select each number to review the corresponding text.CalloutsImmunosandwich (captured and tagged MMB) Excess CrO2 Excess ConjugateThe vessel reactants incubate on the incubate wheel at 37 degrees. During this time an immunosandwich containing capture Ab + sample + tag Ab forms. Incubation times will vary according to the particular HM test being run.8Wash wheel magnets are used to separate the reactants. Select Next to continue.Select each number to review the corresponding text.CalloutsImmunosandwich (captured and tagged MMB)Excess CrO2Excess ConjugateAfter incubation, the reaction vessel is transferred to the wash wheel for magnetic separation. Because chromium dioxide is magnetic, the 12 stationary magnets positioned around the wash wheel pull only the chromium dioxide-containing immunosandwich plus excess chromium dioxide aside.9 Wash probe #1 uses the longer probe to aspirate unbound sample and conjugate. Select Next to continue. Select each number to review the corresponding text.CalloutsImmunosandwich (captured and tagged MMB)Excess CrO2Wash Probe 1 uses the longer, aspirate probe to remove sample and unbound conjugate (excess MMB tag Ab). The only conjugate Ab remaining is that which has bound to the MMB analyte in the sample and is part of the immunosandwich. The amount of bound conjugate Ab is directly proportional to the amount of MMB in the sample. Not all chromium dioxide Ab binds to sample MMB. Remember that excess chromium dioxide Ab still remains in the reaction vessel.10 Wash probe #1 uses the shorter probe to add Chemistry Wash to the reaction vessel. Select Next to continue. Select each number to review the corresponding text.CalloutsImmunosandwich (captured and tagged MMB)Excess CrO2To wash the bound reactants and facilitate removing unbound conjugate Ab, Wash Probe 1 uses the shorter probe to add Chemistry Wash to the reaction vessel.11 The two vessel mixers on the wash wheel resuspend the bound and unbound reactants. Select Next to continue. Select each number to review the corresponding text.CalloutsImmunosandwich (captured and tagged MMB)Excess CrO2The two vessel mixers on the wash wheel resuspend the bound and unbound reactants.12 Wash Cycle Steps Step 1: In the Wash wheel area, stationary magnets separate bound and unbound reactants in the reaction vessel. Step 2: Wash probe aspirates unbound reactants. Step 3: Wash probe dispenses Chemistry Wash reagent to wash the bound reactants and remove unbound reactants.   Step 4: Vessel mixers resuspend the bound and unbound reactants in the reaction vessel. Step 5: HM module repeats this process until four wash cycles have been completed. Select Next to continue. Select each number to review the corresponding text.CalloutsWash probe 2Wash probe 1The reaction vessel now has completed one wash cycle. For each immunoassay, the HM module will complete a total of four wash cycles to remove all excess conjugate. Wash Probe 1 will complete two wash cycles, and then Wash Probe 2 will complete the remaining two wash cycles. 13 The R1 probe delivers reagent from the Flex® reagent cartridge into cuvettes to prepare for the photometric read. Select Next to continue. Select each number to review the corresponding text.CalloutsCPRG added from Flex reagent cartridgeThe R1 probe delivers reagent from the Flex® cartridge into cuvettes to prepare for the photometric read.14 The photometric sample probe transfers the washed immunosandwiches from the reaction vessels to cuvettes. Select Next to continue. Select each number to review the corresponding text.CalloutsImmunosandwich & excess CrO2 added The photometric sample probe transfers the washed immunosandwiches from the reaction vessels to cuvettes.15 In the cuvette, a colorimetric reaction takes place and test results are read photometrically. The color produced is proportional to the amount of analyte in the sample. Select Next to continue. Select each number to review the corresponding text.CalloutsCPRG (non-absorbing at 577nm)CPR color (absorbs at 577nm)In the cuvette, a colorimetric reaction takes place and test results are read photometrically. The color produced is proportional to the amount of analyte in the sample.16 Congratulations. You have learned how heterogeneous immunoassays are performed. When complete select the X in the upper-right corner to close the window and continue.Congratulations. Now that you've learned how heterogeneous immunoassays are performed, let's look at how the HM components function during processing. Now that you’ve learned about basic HM concepts, let us look at key HM components and their functions during HM sample processing. HM Components and Processing Learn how HM components function to process samples. Slide NumberText BlocksCalloutsAudio ScriptImage File1 In this section, you will learn how HM components function to process samples. Select Next to continue. Note: If audio does not automatically start, select the play arrow in the top left to begin.In this section, you will learn about the functions of the HM components in processing samples.2 The vessel holder feeds reaction vessels to the track system where they are loaded onto the incubate wheel by the vessel transfer shuttle. Select Next to continue. Select each number to review the corresponding text. CalloutsVessel HolderTrackIncubate wheelVessel Transfer ShuttleReaction Vessels feed from the vessel holder to the track system. From there, they are loaded onto the incubate wheel by the vessel transfer shuttle.3 The R2 reagent probe picks up and delivers reagents to the reaction vessel. The photometric sample probe picks up and delivers sample to the same reaction vessel. Select Next to continue. Select each number to review the corresponding text.CalloutsPhotometric Sample ProbeR2 ProbeThe R2 reagent probe picks up and delivers reagents to the reaction vessel. The photometric sample probe picks up and delivers sample to the same reaction vessel.4 The incubate wheel incubates reactionvessel contents to allow reactants to form an immunosandwich. The incubate wheel has 45 slots and temperature specifications are 37.3-39.6° C. Incubation times vary according to the particular HM test being run. NOTE: Without the LOCI® module, the temperature specification for the incubate wheel is 42-44° C. Select Next to continue. Select each number to review the corresponding text.CalloutsIncubate WheelReaction VesselThe incubate wheel incubates reaction vessel contents. This allows reactants to form an immunosandwich. The incubate wheel has 45 slots and maintains a temperature of 37-39° C. Incubation times may vary according to the particular HM test being run.5 After incubation, the vessel transfer shuttle moves reaction vessels to the inner wash wheel, where the wash cycle begins by magnetically separating chromium-containing bound reactants from unbound reactants in the reaction vessel. Select Next to continue. Select each number to review the corresponding text.CalloutsVessel Transfer StationWash wheelAfter incubation is completed, the vessel transfer shuttle moves reaction vessels to the inner wash wheel. At this point, the wash cycle begins by magnetically separating chromium-containing bound reactants from unbound reactants in the reaction vessel.6 Wash station probes aspirate unbound reactants and dispense Chemistry Wash reagent into the reaction vessel to further separate bound from unbound reactants. Select Next to continue. Select each number to review the corresponding text.CalloutsIncubate WheelWash WheelWash Probe 2Wash Probe 1Wash station probes aspirate unbound reactants and dispense Chemistry Wash reagent into the reaction vessel, further separating bound from unbound reactants.7 Vessel mixers, located under the wash wheel, resuspend vessel contents. The mixer can process either a single vessel or two vessels simultaneously. Vortex mixing concludes the wash cycle. The HM module repeats the cycle until four wash cycles have been performed. Select Next to continue. Reminder: Magnets are used to separate chromium bound reactants from unbound reactants.Select each number to review the corresponding text.CalloutsVessel MixersMagnetVessel mixers which are located under the wash wheel resuspend vessel contents. The mixer can process either a single vessel or two vessels simultaneously. Vortex mixing concludes the wash cycle. The HM module repeats the cycle until four wash cycles have been performed.8The R1 reagent probe adds Flex® cartridge reagents to a cuvette. The vessel transfer shuttle moves the reaction vessel to the incubate wheel, and the photometric sample probe transfers immunoassay reactants to the same cuvette. Select Next to continue.  Select each number to review the corresponding text.CalloutsIncubate/Wash WheelPhotometric Sample ProbeR1 Reagent ProbeThe R1 reagent probe adds Flex cartridge reagents to a cuvette and the photometric sample probe transfers immunoassay reactants to the same cuvette.9The transfer shuttle moves the used reaction vessels to the waste chute which then discards the vessels to a waste container. Select Next to continue.Avoid Vessel Jams: The waste container liner (not pictured) should be fully open and conform to the inside walls and bottom of the container.The transfer shuttle moves the used reaction vessels to the waste chute which then discards the vessels to a waste container. Make sure the waste container liner is fully open and conforms to the inside walls and bottom of the container. Failure to insert the liner properly may result in vessel jams in the waste chute.10 Congratulations. You have learned how HM components function to process samples. When complete, select the X in the upper-right corner to close the window and continue.Congratulation. You now have learned how HM components function to process samples. The Luminescent Oxygen Channeling Immunoassay (LOCI®) technology measures specialty analytes such as TNI (Cardiac Troponin I), TSHL, and FT4L with increased sensitivity and accuracy. The immunoassay sandwich is prepared on the incubation wheel and measurement takes place in the LOCI® read chamber. Homogeneous immunoassays do not require separation of immune complexes from unbound reactants. After a brief incubation, measurements take place in the LOCI® read chamber.     LOCI® Technolgy Learn about the use of LOCI® technology to detect immune complexes. Checklist TitleChecklist TypeChecklist ContentLOCI® TechnologyHTMLSelect each checkbox to learn more about the use of LOCI® technology to detect immune complexes. LOCI® chemiluminescence represents an important advance in immunochemistry because it pairs a highly sensitive immunoassay with a patented, state-of-the-art detection method.Key LOCI® ReactantsHTML LOCI® chemiluminescence involves the reactants shown above. Key 1. Chemibead 2. Monoclonal antibody 3. Antigen (analyte in sample) 4. Biotinylated antibody 5. Biotin 6. Streptavidin 7. Sensibead  ChemibeadsHTML Non-competitive (sandwich) immunoassays use method-specific synthetic Chemibeads that are coated with monoclonal antibodies specific for the analyte being tested. Key 1. Chemibead 2. Monoclonal antibodyBiotinylated AntibodiesHTML If the media does not automatically start, select the play arrow to begin. Immunoassay reactants include the analyte being tested, which serves as the antigen (Ag), as well as a second monoclonal antibody (Ab) specific for the analyte being tested. Biotin is attached to this antibody-it’s a biotinylated antibody. During incubation at 37° C, Ab + Ag + Ab immune complexes form, where antigen is “sandwiched” between monoclonal antibodies. Key 1. Chemibead 2. Monoclonal antibody 3. Antigen (analyte in sample) 4. Biotinylated antibody 5. BiotinSensibeads + StreptavidinHTML If the media does not automatically start, select the play arrow to begin. Immunoassay reactants also include Sensibeads coated with streptavidin. Sensibeads contain a photosensitive dye. Streptavidin is a small protein with unique biochemical properties, such as its high affinity to bind to biotin. When added to the assay, streptavidin quickly binds to the biotin portion of the biotinylated antibody to form bead-pair immune complexes. Key 1. Chemibead 2. Monoclonal antibody 3. Antigen (analyte in sample) 4. Biotinylated antibody 5. Biotin 6. Streptavidin 7. SensibeadGenerating ChemiluminescenceHTML If the media does not automatically start, select the play arrow to begin. When illuminated at a 680 nm wavelength, Sensibeads generate singlet oxygen, which diffuses into nearby bound Chemibeads and triggers a chemiluminescent reaction. The resulting signal is measured at 612 nm and is proportional to the concentration of analyte being tested. Key 1. LED light source 2. Singlet oxygen released from Sensibead 3. Chemiluminescence generated from Chemibead When complete select the X in the upper-right corner to close the window and continue. Now that you have learned about LOCI® technology, let us look at key LOCI® components and their functions during sample processing   LOCI® Components and Processing Learn how LOCI® components function to process a sample. Slide NumberText BlocksCalloutsAudio ScriptImage File1 In this section, you will learn how the various LOCI® components function to process a sample. Select Next to continue. Note: If audio does not automatically start, select the play arrow in the top left to begin.In this section, you will learn how the various LOCI components function to process a sample.2 The vessel holder feeds reaction vessels to the track system, where they are loaded onto the incubate wheel by the vessel transfer shuttle. Select Next to continue. Select each number to review the corresponding text.CalloutsVessel HolderVessel TrackThe vessel holder feeds reaction vessels to the track system where they are loaded onto the incubate wheel by the vessel transfer shuttle.3 The R2 reagent probe aspirates reagents from a Flex® cartridge and delivers to a reaction vessel. Select Next to continue. Select each number to review the corresponding text.CalloutsIncubate WheelR2 ProbeThe R2 reagent probe aspirates reagents from a Flex cartridge and delivers to a reaction vessel.4 The photometric sample probe aspirates specimen from the sample area and delivers it to the same reaction vessel. Select Next to continue. Select each number to review the corresponding text.CalloutsPhotometric Sample ProbeReaction VesselThe photometric sample probe aspirates specimen from the sample area and delivers it to the same reaction vessel.5 The incubate wheel incubates reaction vessel contents to allow reactants to form an immunosandwich. The incubate wheel has 45 slots and temperature specifications are 37.3 -39.6° C. This maintains a temperature in the liquid in the reaction vessel of 36.5 – 37.5° C. Incubation times vary according to the particular LOCI® test being run. Note:  Without  the LOCI®   module, the temperature specification for the incubate wheel is 42-44° C. Select Next to continue. Select each number to review the corresponding text.CalloutsIncubate WheelReaction VesselThe incubate wheel incubates reaction vessel contents to allow reactants to form an immunosandwich. The incubate wheel has 45 slots and maintains a temperature in the range of 37.3-39.6° C. Incubation times vary according to the particular LOC® Itest being run.6 The LOCI® arm transfers the vessel from incubate wheel to the LOCI® reader. Select Next to continue. Select each number to review the corresponding text.CalloutsIncubate WheelLOCI® ReaderLOCI® ArmThe LOCI® arm transfers the vessel from the incubate wheel to the LOCI® reader.7 When illuminated at a 680 nm wavelength, Sensibeads generate singlet oxygen, which diffuses into nearby bound Chemibeads and triggers a chemiluminescent reaction. The resulting signal is measured at 612 nm and is proportional to the concentration of the analyte being tested. Select Next to continue. Select each number to review the corresponding text.CalloutsLED light sourceSinglet oxygen released from SensibeadChemiluminescence generated from ChemibeadFollowing a flash of light at 680nm, singlet oxygen is produced by the sensibeads. Singlet oxygen is diffused into bound chemibeads, producing chemiluminescence. Light produced is measured at 612nm. Signal produced is proportional to the amount of analyte present.8 Congratulations. You now have learned how key LOCI® components function in processing samples. When complete select the X in the upper-right corner to close the window and continue.Congratulations. You now have learned how key LOCI components function in processing samples. The HM and LOCI® technologies require periodic replacement of consumables. Use the HM System Counters screen to monitor which consumables need replacing. To access this screen from the Operating Menu, select F4: SYSTEM PREP, F6: SYSTEM COUNTERS, F6: HM COUNTERS. Note: From the Daily Maintenance Routines Screen, F3: CHECK HM COUNTS will be populated when there is an HM consumable need. Pressing F3 displays the HM Counter Screen. Refer to the Operator’s Guide for detailed instructions on how to replace HM consumables. Consumables Learn about the HM and LOCI® consumables. Tab TitleTextChemistry Wash Chemistry Wash is used to wash away unbound sample and reagents from the immunosandwich during HM processing. Chemistry Wash is located behind the Pump Panel. Replacewhen there is less than 5% remaining as indicated on the Daily Maintenance Routines screen (F3: Chk HM Counts) or if an error message appears, indicating that the consumable bottle is empty. The volumes of these fluids are monitored by a low-level sensor in each container. Note: To prevent possible contamination, do not pool bottles of Chemistry Wash solution when replacing this consumable.    Reaction Vessels The HM system uses special containers called reaction vessels. The operator loads reaction vessels into the vessel holder, and then the vessel feeder and track system automatically load reaction vessels from the vessel track onto the incubate wheel as needed. When replacing, do not overfill the reaction vessel holder. Filling above the red line may cause jams. Note:  The Dimension® EXL™ with LM integrated chemistry system uses only reaction vessels with a plastic tip on the bottom.    Reagent Probe Cleaner Reagent Probe Cleaner, which contains 0.1N sodium hydroxide, is pumped to one of the R2 probe dual drains and is used to clean the R2 reagent probe and drain for HM processing. Reagent Probe Cleaner is located underneath the film cartridge. Replace Reagent Probe Cleaner when there is less than 5% remaining as indicated on the Daily Maintenance Routines screen (F3: Chk HM Counts) or if an error message appears, indicating that the consumable bottle is empty. The volumes of these fluids are monitored by a low-level sensor in each container.   Sample Probe Cleaner Sample Probe Cleaner, which contains dilute sodium hypochlorite (bleach), fills one of the sample probe dual drains. The photometric sample probe performs this extra rinsing step to ensure that it is thoroughly cleaned to prevent sample carryover. Sample Probe Cleaner is located under the film cartridge. Replace Sample Probe Cleaner when there is less than 5% remaining as indicated on the Daily Maintenance Routines screen (F3: Chk HM Counts) or if an error message appears, indicating that the consumable bottle is empty. The volumes of these fluids are monitored by a low-level sensor in each container.   Flex® Flex® reagent cartridges contain reagents necessary to perform LOCI®(left) and HM (right) testing, as well as others. NOTE:  Note color and lid cover of Flex® reagent cartridges which are used to protect reagents from light. When complete, select the X in the upper-right corner to close the window and continue.   Congratulations. You have completed the Assay Measurement for HM and LOCI® technolgies course for the Dimension® EXL™ with LM integrated chemistry system. Listed below are the key points that have been presented.  Take time to review the material before you proceed to the final quiz.   Describe the HM reaction and processing steps The Heterogeneous Immunoassay Module (HM) is used to process specialty analytes of low molecular weight. The immunoassay is prepared in the HM area, and the final results are read photometrically after transferring reactants from reaction vessels to cuvettes. The vessel holder feeds reaction vessels to the track system, where they are loaded onto the incubate wheel. The R2 reagent probe aspirates and delivers reagents to the reaction vessel. The photometric sample probe aspirates and delivers sample to the same reaction vessel.   The incubate wheel has 45 slots and maintains a temperature of 37° C in the reaction vessel to allow reactants to form an immunosandwich. Incubation times vary according to the particular HM test being run. The vessel transfer shuttle moves the reaction vessel from the incubate wheel to the wash wheel where immunosandwiches are magnetically separated from unbound reactants in the vessel. Once the reaction vessel is shuttled back to the incubate wheel, the content is transferred from the reaction vessel to a cuvette for measurement.   Describe the homogenous processing using LOCI® technology The Luminescent Oxygen Channeling Immunoassay (LOCI®) technology measures specialty analytes such as TNI (Cardio Troponin I), TSHL, and FT4L with increased sensitivity and accuracy. The immunoassay sandwich is prepared on the incubation wheel and measurement takes place in the LOCI® read chamber. Homogeneous immunoassays do not require separation of immune complexes from unbound reactants.After a brief incubation, measurements take place in the LOCI® read chamber. The vessel holder feeds reaction vessels to the track system, where they are loaded onto the incubate wheel. The R2 reagent probe aspirates and delivers reagents to the reaction vessel. The photometric sample probe aspirates and delivers sample to the same reaction vessel.  The incubate wheel incubates reaction vessel content to allow reactants to form an immunosandwich. The incubate wheel has 45 slots and maintains a temperature of 37° C. Incubation times vary according to the particular LOCI® test being run. The LOCI® arm transfers the reaction vessel from the incubate wheel to the LOCI® reader. Following a flash of light at 680nm, singlet oxygen is produced by the sensibeads. Singlet oxygen is diffused into bound chemibeads, producing chemiluminescence. Light produced is measured at 612nm. The signal produced is proportional to the amount of analyte present.   Identify key components and their functions for HM and/or LOCI®testing Both HM and LOCI® testing use the following components:       R2 reagent probe is used to transfer reagents from Flex® cartridges to reaction vessels Photometric Sample Probe is used to aspirate and dispense sample into reaction vessels In addition, HM uses the following components:   The HM wash probes are used to wash away unbound material from the reaction vessel The Sample Probe transfers reactants from the reaction vessel to a cuvette Cuvettes hold final HM reactants and are read photometrically by the photometer   Identify consumables used by the HM and LOCI®systems Chemistry Wash washes unbound sample and reagents from the immunosandwich, which is made up of the analyte being measured + chromium-based capture antibody + tag antibody. Replace Chemistry Wash when there is less than 5% remaining as indicated on the Daily Maintenance Routines screen (F3: Chk HM Counts) or if an error message appears, indicating that the consumable bottle is empty. The volumes of these fluids are monitored by a low-level sensor in each container. To prevent possible contamination, do not pool bottles of Chemistry Wash solution when replacing this consumable. Chemistry Wash is located behind the Pump Panel. The HM system uses special containers called reaction vessels. Reactants in the heterogeneous immunoassay are mixed, incubated, and washed in the reaction vessels. The operator loads reaction vessels into the vessel holder, and then the vessel feeder and track system automatically load reaction vessels from the vessel track onto the incubate wheel as needed. When replacing reaction vessels, do not overfill the reaction vessel holder-filling above the red line may cause jams. Reagent Probe Cleaner, which contains sodium hydroxide, fills one of the R2 probe dual drains and is used to clean the R2 reagent probe. Reagent Probe Cleaner is located under the cuvette film cartridge. Select Next to continue.