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Berthold LB 9510 Operating Manual

Lumat luminescence tube luminometer
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LB 9510
Lumat Luminescence Tube Luminometer
Operating Manual
71450BA2
Rev. No .: 00, 12/2020

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Summary of Contents for Berthold LB 9510

  • Page 1 LB 9510 Lumat Luminescence Tube Luminometer Operating Manual 71450BA2 Rev. No .: 00, 12/2020...
  • Page 3 The information in this guide is subject to change without notice. DISCLAIMER TO THE EXTENT ALLOWED BY LAW, BERTHOLD TECHNOLOGIES AND/OR ITS AFFILIATE(S) WILL NOT BE LIABLE FOR SPECIAL, INCIDENTAL, INDIRECT, PUNITIVE, MULTIPLE, OR CONSEQUENTIAL DAMAGES IN CONNECTION WITH OR ARISING FROM THIS DOCUMENT, INCLUDING YOUR USE OF IT.

  • Page 5: Table Of Contents

    LB 9510 Lumat Table of Contents Table of Contents Prefatory Comments ................7 Contact Information .................. 7 Typographical Conventions ............... 8 Safety Manual and Operating Manual ............9 1.3.1 The Operating Manual ................9 Software Installation ................ 10 Installation and Registration of ICE Operating Software ......10 Installation of Driver Software ..............
  • Page 6 Table of Contents LB 9510 Lumat Maintenance of the Injector System ............64 5.1.1 Priming the tubing .................. 64 5.1.2 Customizing the Priming Sequence ............65 5.1.3 Emptying Tubings ..................67 5.1.4 Cleaning the Tubings ................68 Further maintenance information ............69 Quality Control Check ...............

  • Page 7: Prefatory Comments

    LB 9510 Lumat Prefatory Comments Prefatory Comments Contact Information Berthold Technologies GmbH & Co. KG Calmbacher Str. 22 75323 Bad Wildbad Germany Sales and Customer Service Internet: www.berthold.com Phone: +49 7081 177-0 Fax: +49 7081 177-100 E-Mail: bio@berthold.com Central Customer Service...

  • Page 8: Typographical Conventions

    1 Prefatory Comments LB 9510 Lumat Typographical Conventions j DANGER Indicates an imminent, major hazard, which will certainly result in seri- ous injuries or even death if the hazard is not avoided. j CAUTION Refers to a potentially dangerous situation, which can result in me- dium or minor physical injuries or damages to property, if it is not avoided.

  • Page 9: Safety Manual And Operating Manual

    Manual and both before first use of the instrument. 1.3.1 The Operating Manual The operating manual is valid from the delivery of the Berthold product to the user until its disposal. Keep the operating manual for future refer- ence. Read all instructions thoroughly and completely before working with the product.

  • Page 10: Software Installation

    2 Software Installation LB 9510 Lumat Software Installation The instrument is operated with ICE software. Follow the instructions for ICE software installation. IMPORTANT Precondition for both software packages: Windows compatible PC with Windows 10. Installation and Registration of ICE Operating Software...
  • Page 11 LB 9510 Lumat Software Installation IMPORTANT Additional resources: As the software requires some additional resources for proper opera- tion the set up wizard will check for the presence of these resources (.NET Framework 4.5 or newer and Crystal Reports for .NET Frame- work) on the computer.

  • Page 12: Installation Of Driver Software

    2 Software Installation LB 9510 Lumat Alternatively request the password via the BERTHOLD TECHNOLOGIES homepage. Please provide the System ID of the PC. The number is printed in red in the registration dialog of the software. Owner will receive the registration password from BERTHOLD TECHNOLOGIES only.
  • Page 13 LB 9510 Lumat Software Installation Make sure the instrument´s power switch is in OFF position. Insert the USB stick into a USB slot and browse to the USB root direc- tory. The software installation does not start automatically. The USB stick contain a folder with the instrument driver software.

  • Page 14: Adjusting The Response Of Both Adapters

    2 Software Installation LB 9510 Lumat Select the [ComPort]. Select the entry starting with BT20…. The raw data are usually displayed as RLU representing the total amount of counts acquired during the reading time per well. By checking [Calculate RLU/s] the total amount of counts will be di- vided by the respective reading time.

  • Page 15: Instrument Control And Evaluation Software (Ice)

    LB 9510 Lumat Instrument Control and Evaluation Software (ICE) Instrument Control and Evaluation Software (ICE) ICE Directories and Files The directories for data and parameter files are defaulted as described below. Any accessible directory on the computer and the local network can be selected though when saving data and parameter files using the “Save …...

  • Page 16: Ice User Interface

    3 Instrument Control and Evaluation Software (ICE) LB 9510 Lumat ICE User Interface 3.2.1 ICE Default Setup Default main screen of ICE: To return to the default layout after any changes you may have to check [View]/[Navigation panel] and [Lumat].

  • Page 17: File Menu

    LB 9510 Lumat Instrument Control and Evaluation Software (ICE) 3.2.2 File Menu The [File] menu contains commands to open and save data and protocol files. [New] clears data display to start a new measure- ment [Open Protocol File…] opens an existing protocol...

  • Page 18: View Menu

    3 Instrument Control and Evaluation Software (ICE) LB 9510 Lumat 3.2.3 View Menu The View menu defines how the user interface and data are displayed. [Navigation Panel] shows/hides navigation panel on the left [Lumat] adjusts user interface for Lumat [Protocol Wizard]...

  • Page 19: Tools Menu

    LB 9510 Lumat Instrument Control and Evaluation Software (ICE) [Properties] includes instrument driver settings and instru- ment configuration [Load Tube] turns sample rotor once: moves tube from loading position into measurement position [Prime] Opens the injector prime sequence. See chap- ter 5.1.1 for details.

  • Page 20: Help Menu

    3 Instrument Control and Evaluation Software (ICE) LB 9510 Lumat 3.2.6 Help Menu The [Help] menu allows you to view basic software information. Page 20 of 79 71450BA2 Rev. 00, 12/2020...

  • Page 21: Operation With Ice

    LB 9510 Lumat Operation with ICE Operation with ICE Defining protocols and running measurements on the Lumat is straight forward. The procedure is the same for all types of assay types, e.g. Raw Data, Curve Fit and Cut-off. A measurement can be carried out immediately after a stored protocol is selected.

  • Page 22: Define A Raw Data Protocol

    4 Operation with ICE LB 9510 Lumat • Click [Edit] to edit an existing protocol file. The [ReRun&Edit] function is mainly interesting for Curve fit measurements. It allows to re-evalu- ate stored data with an edited parameter file. See chapter 4.8 for further information.
  • Page 23 LB 9510 Lumat Operation with ICE content, single reporter gene expression, ac- tivities of caspases, kinases and many other enzymes. [Endpoint Dual] In a Dual Label Assay two consecutive End- point measurement series are defined. Each series may start with an injection and/or a de- lay.
  • Page 24 4 Operation with ICE LB 9510 Lumat Example for Endpoint Single Insert the following parameters for your Endpoint Single Raw Data protocol in the [General Page]. [Protocol name] enter a descriptive name [Delay time] insert the delay time before the next opera- tion step (0-600s).
  • Page 25 LB 9510 Lumat Operation with ICE Example for Kinetic Single Insert the following parameters for your Kinetic Single Raw Data pro- tocol in the [General Page]. [Protocol name] enter a descriptive name [Total time time] insert the total time of the Kinetic measure- ment (0-86400 s).
  • Page 26 4 Operation with ICE LB 9510 Lumat In case of Endpoint Dual and Dual Glo measurement types the [Mul- tilabel Calculation Page] appears additionally. Select the type of calculation. Click <Next>. Define the parameter for each required injector in the [Injector Page].
  • Page 27 LB 9510 Lumat Operation with ICE The [Measuring Sequence Page] shows the sample and replicate se- quence to be measured. Click <Next>. Define [Print] settings. Select the items by dragging from left to right. [Page Header] file names [Measurement Data] readings...
  • Page 28 4 Operation with ICE LB 9510 Lumat Type your [Footer] specific for this protocol. Define the [Directory] for the export file. Check if [Automatic Export] is required. Click <Next>. Click <Finish> to save the protocol. Page 28 of 79 71450BA2 Rev. 00, 12/2020...

  • Page 29: Perform A Raw Data Measurement

    LB 9510 Lumat Operation with ICE Perform a Raw Data Measurement The steps to perform a Raw Data Measurement are valid for all Raw Data measurement types (Endpoint Single/Dual, Kinetic Single etc.). NOTICE Use of injectors: If injectors are to be used, they must be primed prior to protocol start.

  • Page 30: The Kinetics Tab

    4 Operation with ICE LB 9510 Lumat Example for a DualGlo Assay Click on the tab <Statistics> to view information about your protocol and instrument. In case of kinetic measurements an additional <Kinetics> tab, de- scribed in chapter 4.2.1., is provided. Click on <Kinetics> to view the curves.

  • Page 31: Define A Curvefit Protocol

    LB 9510 Lumat Operation with ICE Define a Curvefit Protocol IMPORTANT Protocol wizard The protocol setup is guided through a wizard. Most of the steps passed through the protocol setup are similar in all kinds of protocols, Raw Data, Cut-Off and Curve Fit.
  • Page 32 4 Operation with ICE LB 9510 Lumat IMPORTANT Description of curvefit protocols As curvefit protocols differ strongly dependent on the selected type of standardization, the type of curve fitting and special requirements of the respective assay system, the provided functions are described in general terms, but without sequence of actions for each standardiza- tion type.
  • Page 33 LB 9510 Lumat Operation with ICE measurement, there will be first shaked and afterwards the instrument is waiting for delay time to be expired. [Measuring time] define your measuring time per well (0.1-600s) [Concentration Unit] Enter the concentration unit to be shown in the report.
  • Page 34 4 Operation with ICE LB 9510 Lumat Insert the following parameters for your Immunoassay protocol in the [Curve Properties Page]. [No. of controls] Use controls to verify your results against known concentrations. Enter the number of controls (0-10). [No. of decimals] Enter the number of decimals (0 to 3) to be shown in the report.
  • Page 35 LB 9510 Lumat Operation with ICE [Type of transformation] select the axis transformation; availability of transformation types is dependent on the selected type of curve fitting. For spline:[logit/log], [log/log] Choose whether to perform a logit/log or log/log transformation prior to the spline fit.
  • Page 36 4 Operation with ICE LB 9510 Lumat above the limit, a warning message will ap- pear in the report. Click <Next> The [Controls Page] of Immunoassay protocol will only appear when controls are defined in the [Curve Properties Page]. Insert the ac- ceptable lower and upper limits (calculated concentration values) for the controls.
  • Page 37 LB 9510 Lumat Operation with ICE Example for a Mastercurve with calibrators and controls [No of standards] Insert the number of standards. Minimum of standards for Spline calculation: 2 Point to point calc: 4 Maximim of standards: 10 [Concentrations] Enter the respective concentration of each standard.
  • Page 38 4 Operation with ICE LB 9510 Lumat [Expected Range] Tic the checkbox to activate the submenu and insert the limit values. Below the low limit and above the high limit, concentrations will not be calculated. [Low Limit][High Limit] Insert a low and a high limit to define your expected range for measurement.
  • Page 39 LB 9510 Lumat Operation with ICE Insert the following parameters: [Volume] insert a volume of 10-100µl. [Speed] select the injection speed (1-5). Speed 2 is preset. [Delay] insert the delay time after injection. The [Sequence] of your defined settings for measurement and injec- tion are shown on the right side of the page.
  • Page 40 4 Operation with ICE LB 9510 Lumat [Sample] sample information [Read] single measured value [Concentration] calculated concentration of a single measured value [Mean Read] mean value of the measured sample repli- cates [Mean conc] mean calculated concentration of the sample replicates [c.v.]...

  • Page 41: Perform A Curvefit Measurement

    LB 9510 Lumat Operation with ICE Perform a Curvefit Measurement Depending on the type of standardization, the steps to be performed vary for the different types of Immunoassays . See chapter 9.1 for details on the mathematical data reduction used in the software after measurement of a full standard curve.
  • Page 42 4 Operation with ICE LB 9510 Lumat Insert the 1st tube, containing the first replicate of the lowest stand- ard according to the measuring sequence. Record a comment if desired. Click <OK> on the [Start Dialog] screen to start the measurement. The tube will be measured and the numerical value of the signal as well as the concentration will be displayed in the <Data>...

  • Page 43: Measure An Immunoassay With Reference Curve

    LB 9510 Lumat Operation with ICE Standard curve tab of measurement A right-click into the chart area opens a menu offering display, print and save options. Curve fit method, axis transformation and smoothing may be changed and values of standards may be edited if appropriate. The edited and recalculated data will be indicated by red colour.
  • Page 44 4 Operation with ICE LB 9510 Lumat To check the protocol settings you may click <Protocol Preview>. Click <Measure Ref-Std Curve>. The standards for your reference curve, underlaid with green color, will be displayed according to your definitions. Click <Start Measurement>. A [Start Dialog] screen will be displayed.

  • Page 45: Measure An Immunoassay With Master Curve

    LB 9510 Lumat Operation with ICE Enter a [file name] under which the measurement is to be stored. Follow the description of 4.-13. in chapter 4.4.1, select a [Reference curve] and measure the 1 replicate of calibrator 1 and the following tubes according to the given sequence.
  • Page 46 4 Operation with ICE LB 9510 Lumat A right-click into the chart area opens a menu offering display, print and save options. Curve fit method, axis transformation and smoothing may be changed and values of standards may be edited if appropriate.

  • Page 47: Define A Cut-Off Protocol

    LB 9510 Lumat Operation with ICE Define a Cut-Off Protocol IMPORTANT Protocol wizard The protocol setup is guided through a wizard. Most of the steps passed through the protocol setup are similar in all kinds of protocols, Raw Data, Cut-Off and Curve Fit.
  • Page 48 4 Operation with ICE LB 9510 Lumat [Delay time] insert the delay time before the next opera- tion step (0-600s). [Measuring time] Insert your measuring time (0.1-600s) [Replicates of samples] insert the number of replicates [Perform Background Measurement] Tic the checkbox to activate the submenu.
  • Page 49 LB 9510 Lumat Operation with ICE Define the [Offset] (AL): a fixed value [High Cut Off] i.e. the RLU value above which a result will be flagged positive. Define the [Offset] (AH): a fixed value Applied formulas: Low cut-off: AL...
  • Page 50 4 Operation with ICE LB 9510 Lumat Define the [Offset] (AL): a fixed value Define the [Factor] (BL): a factor to determine the negative control’s weight on the low cut- off. [High Cut Off] i.e. the RLU value above which a result will be flagged positive.
  • Page 51 LB 9510 Lumat Operation with ICE Use the settings as described below when • a positive control is measured besides the samples: [No. of positive controls] 1-3 [Replicates of Positive control] [Low Cut Off] i.e. the RLU value below which a result will be...
  • Page 52 4 Operation with ICE LB 9510 Lumat [%CV of ……] Define the max. acceptable coefficient of var- iation of the replicates of the samples and the negative control, above which the results will be flagged. Use the settings as described below when •...
  • Page 53 LB 9510 Lumat Operation with ICE Define the [Lpos factor] (CH): a factor to de- termine the positive control’s weight on the high cut-off. Applied formulas: Low cut-off = AL + BL*NegContr + CL*PosContr High Cut-Off = AH + BH*NegContr + CH*PosContr [%CV of ……]...
  • Page 54 4 Operation with ICE LB 9510 Lumat [use negative control] checked or unchecked. The negative control may be used for quality control. [Ratio Cut-Off] checked [No. of positive controls] Set the number of pos. controls to 1 or 2. Only positive control 1defines the cut-off.
  • Page 55 LB 9510 Lumat Operation with ICE [%CV of ……] Define the max. acceptable coefficient of var- iation of the replicates of the samples and the controls, above which the results will be flagged. [Low RLU of negative control] Define the limit of the RLU value be- low which the negative control will be flagged.
  • Page 56 4 Operation with ICE LB 9510 Lumat Define the parameter for each required injector in the [Injector Page]. This page is only available if the instrument is equipped with injectors. Insert the following parameters: To activate an injector, click the respective injector button and tic the checkbox [use injector x].
  • Page 57 LB 9510 Lumat Operation with ICE Define the settings for printout in the [Print Page]. Select the items by dragging from left to right. [Page Header] file names [Measurement Data] readings [Statistics] measurement settings Define [page orientation] and [margins]. Check if [Automatic Print after Measurement] is required.

  • Page 58: Perform A Cut-Off Measurement

    4 Operation with ICE LB 9510 Lumat Perform a Cut-off Measurement The steps to perform a Cut-Off Measurement are valid for all types of Cut- off protocols. NOTICE Use of injectors: If injectors are to be used, they must be primed prior to protocol start.

  • Page 59: Data Export And Print Out

    LB 9510 Lumat Operation with ICE When the measurement has finished, remove the last sample (still in measurement position) from the instrument using [Instrument] / [Load Tube]. Data export and print out The export and print-out of results are accessible through the report function of ICE software.

  • Page 60: Direct Data Print-Out

    4 Operation with ICE LB 9510 Lumat 4.7.2 Direct data print-out If not opened already, open the respective file by selecting [File] / [Open data file]. Select [File] / [Print preview] to get a preview of your print-out. Select [File] / [Print] or click on the printer icon in the left-hand navi- gation bar to start printing the data.

  • Page 61: Data Print-Out Via Report

    LB 9510 Lumat Operation with ICE Click the <Export> icon on the left side of the [Report form]. • Select the [target directory] • Define the [file name] • Select the [file type] Adobe Acrobat – saves report in pdf format Excel –...

  • Page 62: Rerun & Edit

    4 Operation with ICE LB 9510 Lumat ReRun & Edit This function is mainly interesting for Curve fit measurements. It allows to re-evaluate stored data with a parameter file, e.g. with another fit, an- other set of calibrators or to get additional copies of print-outs or export files.

  • Page 63: Editing Data

    LB 9510 Lumat Operation with ICE The use of latest calibrators will be indicated by an exclamation mark. You may now print or export your results. 4.8.2 Editing Data Data that have been opened through [ReRun & Edit] can be edited if nec- essary –...

  • Page 64: Maintenance

    For good operation, the injector lines have to be maintained regularly and must be primed previously and washed after use. • Berthold Technology recommends strongly to perform the priming with deionized water first and leaving the lines filled with deionized...

  • Page 65: Customizing The Priming Sequence

    LB 9510 Lumat Maintenance Select the prime mode. Check [Use Customized Prime] to select a user defined priming method. See chapter 5.1.2 for further information. Uncheck [Use Customized Prime] to use the default prime mode. Select the [injectors]. All injectors may be selected at the same time.
  • Page 66 5 Maintenance LB 9510 Lumat Available settings: Injection: the injector is filled with the max. injection volume from the reagent reservoir and injects the set volume Volume percentage of the max. inj. vol. Delay delay before the operation in sec Speed 1 …...

  • Page 67: Emptying Tubings

    LB 9510 Lumat Maintenance dragging it from the left column [Operations] to the right column [Se- quence]. To change the sequence, use the arrow buttons on the right side. To remove an operation, click on it and use the dustbin buttons.

  • Page 68: Cleaning The Tubings

    Enter the [Number of Wash Cycles]. One wash cycle consists of one shot. 10 cycles ensure that the injector system is completely filled. Depending on the substances used, Berthold Technologies recom- mends up to 30 cycles for washing a single injector.

  • Page 69: Further Maintenance Information

    LB 9510 Lumat Maintenance Further maintenance information Further information on maintenance items can be found in the Safety Manual: Safety Manual Chapter Item Replace tubing connections Special spare parts Replace reagent filters 5.4 and sub-chapters Cleaning and Decontamination Disposal 71450BA2 Rev. 00, 12/2020...

  • Page 70: Quality Control Check

    It is recommended to run known internal quality standards or samples attendant to the measurement runs or to use the Luminescence Test Tube, offered by Berthold Technologies. Page 70 of 79 71450BA2 Rev. 00, 12/2020...

  • Page 71: Warranty And Technical Issues

    • the product has not been regularly and correctly maintained • the product has not been repaired or modified by a Berthold Tech- nologies authorized representative or user • parts other than original Berthold Technologies parts are used •...

  • Page 72: Customer Service

    Customer Service Customer service will be provided in the first instance by the network of Berthold Technologies representatives. In the event of any problem ex- perienced with your instrument, the first recourse should be your local Berthold Technologies representative. For further problems requiring hardware or software expertise, the Technical Support group at Berthold Technologies GmbH &...

  • Page 73: Technical Data Of Ice Software

    LB 9510 Lumat Technical Data of ICE Software Technical Data of ICE Software Operating system Win 10 (32/64bit) PC requirements Pentium like CPU (2 GHz or better/intel Core iX recommended), 1 free USB 2.0 port. Interface(s) ICE Software Wizard support for parameter entries...

  • Page 74: Appendix

    9 Appendix LB 9510 Lumat Appendix Mathematical Data Reduction This appendix provides an overview of the LIA data reduction used in the software, which is performed after the measurement of a full standard curve. The following steps take place one after another: The ZERO concentration factor f is calculated if the first standard con- centration = 0.
  • Page 75 LB 9510 Lumat Appendix – NSB) / (B – NSB) where NSB could also be = 0, i.e. for all concentrations C in c in b = log (f * C = logit (B – NSB) / (B – NSB)
  • Page 76 9 Appendix LB 9510 Lumat This factor is determined according to the formula the limits 1 <f < 100 are valid for f. As a consequence, the curve will be fairly linear at the zero point. Spline Function The spline .
  • Page 77 LB 9510 Lumat Appendix does not meet the specified requirements, the standard curve is consid- ered incorrect and the user may have to correct the individual values. Please Note: Before calculating the spline curve, the standards are checked for strict monotony (rising or falling, according to the curve type).
  • Page 78 9 Appendix LB 9510 Lumat plausible substitution values for the maximum binding (either 20 % or nearly 100%). If in special cases only two value pairs are available for calculation, the slope factor is equated with 1. Extrapolation of the Concentration...

  • Page 79: Index

    LB 9510 Lumat Appendix Index 4 parameter logistic ......74 ICE Software Installation ....10 Adjust adapter response ....14 ICE Technical Data ......73 Cleaning the Tubings ......68 ICE User Interface ......16 Confirmation on Decontamination ..72 Immunoassay ........

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