LUMAT LB 9507 User manual

LUMAT
LB 9507
Variable Injector
Firmware Version 5.03
Id. No.: 81 957 BA4
Rev. No.: 06 5.03.2001

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LB 9507 Contents
i
Contents
Trademarks iv
1. SYSTEM DESCRIPTION 1-1
1.1 Application 1-1
1.2 Structure and Operation of the Software 1-3
1.3 The Principle of Measurement 1-8
1.4 Sample Tubes 1-9
1.5 Data Management and Software update 1-10
1.6 Simple and Reliable Operation 1-11
1.7 Instrument and software versions 1-11
2. DESIGN OF LUMAT LB 9507 2-1
2.1 Keyboard 2-4
2.2 Display 2-5
2.3 Printer 2-5
2.4 Measurement Unit 2-6
2.5 Reagent Storage Area 2-10
2.6 Variable injectors 2-11
2.7 Reagent Supply 2-13
2.8 Memory Management 2-14
3. GETTING STARTED 3-1
3.1 Instrument Setup 3-1
3.2 Power Supply 3-1
3.3 Checking the Paper Supply 3-1
3.4 Switching on Lumat 3-2
3.5 System Configuration Parameters 3-3
3.5.1 Description of the System Configuration Parameters 3-4
3.5.2 System Configuration Table 3-6
3.6 Instrument Parameters 3-7
3.6.1 Setting/Verifying the Instrument Parameters 3-7
3.6.2 Changing the Instrument Parameters 3-9
3.7 Reagent Supply 3-11
3.8 Safety Precautions for Operation 3-13
4. STRUCTURE AND OPERATION OF SOFTWARE 4-1
4.1 Operation 4-1
4.2 The Six Functions of the Main Menu 4-2
4.2.1 <MEASURE> Menu 4-3
4.2.2 <PROTOCOL> Menu 4-4
4.2.3 <OPERATOR FUNCTIONS> Menu 4-6
4.2.4 <SYSTEM CONFIGURATION> Menu 4-7
4.2.5 <INSTRUMENT PARAMETER> Menu 4-8
4.2.6 <TURN ONCE> function 4-8
4.2.7 Service functions 4-8
4.3 General Procedure 4-9
5. RAW DATA MEASUREMENTS 5-1

Contents LB 9507
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5.1 Introduction 5-1
5.2 Creating Raw Data Protocols 5-3
5.3 Editing/Deleting Protocols 5-9
5.4 Performing Raw Data Measurements 5-11
6. CUT-OFF MEASUREMENTS 6-1
6.1 Introduction 6-1
6.2 Creating CUT-OFF Protocols 6-4
6.3 Editing/Deleting Protocols 6-12
6.4 Running Cut-off Measurements 6-14
7. QUANTITATIVE MEASUREMENTS (LIA...) 7-1
7.1 Introduction 7-1
7.2 Setting Up Quantitative Protocols 7-3
7.3 Editing/Deleting Protocols 7-12
7.4 Entering a Master Curve 7-14
7.5 Performing Quantitative Measurements 7-17
7.5.1 Measurement with a Complete Standard Curve 7-17
7.5.2 Measurements with Master Curve 7-25
8. T-UPTAKE/FTI-MEASUREMENTS 8-1
8.1 Introduction 8-1
8.2 Creating T-uptake/FTI protocols 8-3
8.3 Editing/Deleting protocols 8-12
8.4 Performing T-uptake/FTI Measurements 8-14
9. DUAL-LUCIFERASETM REPORTER GENE ASSAY 9-1
9.1 Creating Protocols for Dual-LuciferaseTM Assay 9-3
9.1.1 With 2 injectors installed 9-3
9.1.2 With 1 injector installed 9-8
9.1.3 Without injectors 9-11
9.2 Editing/Deleting Protocols 9-14
9.3 Performing Dual-LuciferaseTM Reporter Gene Assay
Measurements 9-16
10. KINETIC AND REPEATED MEASUREMENTS 10-1
10.1 Kinetic Option in Raw Data Protocol 10-1
10.2 Repeated Measurements 10-2
10.2.1 Creating Repeated Protocols 10-4
10.2.2 Editing/Deleting Protocols 10-9
10.2.3 Performing Repeated Measurements 10-11
10.3 Ratemeter 10-14
11. OPERATOR FUNCTIONS 11-1
11.1 <REAGENT> Function 11-2
11.1.1 Loading reagents 11-3
11.1.2 Refreshing reagents 11-4
11.1.3 Washing of the injector system 11-5
11.1.4 Removing reagents 11-7
11.2 Performance test 11-8
11.3 Manual Data Entry 11-9

LB 9507 Contents
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11.4 Retrieval of Stored Data 11-10
11.4.1 Rerun of a Full Standard Protocol 11-11
11.4.2 Rerun of a 2-Point Calibrator Protocol 11-14
12. CONNECTION TO EXTERNAL COMPUTERS 12-1
12.1 General 12-1
12.2 RS 232 Data Output LB 9507 in Protocol-Specific
Measuring Mode 12-2
12.3 Data Output in Raw Data Measurements 12-4
12.4 Data Output after Manual Input of RLU Values 12-4
12.5 Data Output of Stored Assay Data 12-4
12.6 Data Output of RLU/s in the Ratemeter Mode 12-4
13. MAINTENANCE 13-1
13.1 Supplying Paper 13-1
13.2 Replacing the Mains Fuse 13-2
13.3 Changing the Operating Voltage 13-3
13.4 Cleaning the Injector System 13-3
13.5 Cleaning Lumat 13-3
13.6 Changing the Air Filter 13-3
13.7 Checking the Hose Connections 13-3
14. TROUBLESHOOTING 14-1
14.1 Error Messages during Operation 14-1
14.2 Injector System Malfunction 14-1
14.3 Other Errors 14-2
14.4 Default Values 14-2
15. TECHNICAL SPECIFICATIONS OF LB 9507 15-1
16. FLOW DIAGRAM OF THE MEASUREMENT
PROCEDURE 16-1
APPENDIX 1
A.1 Mathematical Basis 2
A.1.1 Overview of the Mathematical Data Reduction 2
A.1.2 Variable Zero Concentration Factor 4
A.1.3 Spline Function 5
A.1.4 Calculation of a Variable Reference Value 6
for Standardization of the Logit Transformation 6
A.1.5 Extrapolation of the Concentration 7
A.2 Data Acquisition of the Lumat LB 9507 8
via the Serial Interface (I/O-Port) and 8
Saving this Data in a MS-DOS ASCII-File 8
A.3 Connection Cable Lumat/Apple Macintosh Computer 9
A.4 Background measurement 10
A.5 Overload Detection 12
A.6 Glossary 13
A.7 Index I

Contents LB 9507
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Trademarks
Dual-Luciferase and Stop and Glow are trademarks of Promega Corp.
Lumat is a trademark of Berthold Technologies.
Macintosh is a registered trademark of Apple Computer Inc.
Excel is a registered trademark of Microsoft Corp.

LB 9507 Preface
I
Preface
Use and Function The Lumat LB 9507 is a semi-automatic luminometer for universal
use in bio- and chemiluminescence, and is very easy to operate. It
can work with up to 2 reagent injectors, and its rotating sampling
method enables samples to be measured and loaded at the same
time.
Special Instructions
and Precautions The manufacturer has undertaken everything to ensure the safe
operation of the instrument (regarding electricity, electronics and
mechanics). The user has to make sure that the equipment is
set up and installed properly to guarantee safe operation:
•Use grounded mains.
•Do not open the instrument.
•Disconnect Lumat from the mains before cleaning.
•If the operational safety of the instrument is no longer ensured,
turn it off and disconnect it from the mains.
•If liquid gets into the instrument, disconnect it from the mains.
•Protect yourself from electrostatic charging (e.g. carpets),
since discharging may damage sensitive electronic
components
•Set up the instrument in a dry room only.
•Be careful when handling strong alkaline or acidic reagents.
Make sure you follow the reagent manufacturer’s instructions.
•If there is a leak in the reagent tubing system you must abort
the measurement series and fix the leak. If necessary, call the
manufacturer’s Technical Service. If a leak occurs there is a
risk of reagent entering the inside of the instrument.
•Only technicians authorized by Berthold should carry out
repairs or modifications on the instrument.
The Lumat LB 9507 should only be used to perform the
functions described in this manual.
The proper use and handling of the instrument requires that the
operating personnel be familiar with this manual. Please read it
carefully before taking the instrument into use - even though the
operation of Lumat may appear simple.

Preface LB 9507
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The tests and maintenance routines recommended by the
manufacturer should be carried out to ensure the operator’s safety
as well as the correct performance of the instrument.
All maintenance and service routines above and beyond those
mentioned in this Operator’s Manual must only be performed by
service technicians authorized by BERTHOLD TECHNOLOGIES
or by the company itself.
The Structure of
the operating manual Because the Lumat LB 9507 is an universal luminometer, it is
capable of a wide range of applications and different
measurement modes (raw data, cut-off, kinetics, repeated, T-
Uptake/FTI, Dual-LuciferaseTM reporter gene assay and
quantitative measurements). To make it easy for the user who is
working with only one measurement mode to find the required
information quickly, this operating manual includes all the relevant
information on each measurement mode in separate chapters.
This means you will not need to search through the manual to find
a different measurement mode.
The description of each measurement mode follows the same
pattern: first a brief introduction, then a flow chart providing a
summary of the procedure, and finally a detailed explanation of
how to create a protocol and run a measurement, using all the
messages shown on the instrument’s display.
Chapter 1 describes the system construction, the ways it can be used, and
the operating procedure
Chapter 2 provides detailed information on the individual system
components
Chapter 3 describes how to commission Lumat, from assembly to
configuration
Chapter 4 explains the structure and handling of the software and its six
menus
Chapter 5 describes the Raw data measurement mode
Chapter 6 describes the Cut-off measurement mode
Chapter 7 describes the Quantitative measurement mode (LIA..)
Chapter 8 describes the T-Uptake/FTI measurement mode
Chapter 9 describes the Dual-LuciferaseTM Reporter Gene Assay mode
Chapter 10 describes the Kinetic and Repeated measurement mode

LB 9507 Preface
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Chapter 11 describes the special instrument functions such as washing,
pumping back reagents, performance testing, etc.
Chapter 12 explains how to connect an external PC and how to transfer data
Chapter 13 summarizes the maintenance and service routines that can and
should be carried out by the user
Chapter 14 lists all possible error messages and tells you how to eliminate
problems
Chapter 15 lists technical specifications of Lumat
Chapter 16 contains a diagram and a flow chart to help you find your way
around the menu structure in the Lumat software. The diagram
shows the assignment of the functions to the menus, and the flow
chart shows the procedure when creating measurement protocols
Appendix includes a description of the mathematical basis for calculations, a
glossary and an index to help you find your way around, and finally
the data sheets.
Writing conventions The description of the software and dialogs are based on a writing
convention, which uses repeated symbols and makes reading
easier.
The software dialog, i.e. the input sequences performed by the
user via keyboard and display, are described
a) by illustrating the displays one after another
b) by explanatory texts.
How displays are shown In the displays shown, the functions in bold face are those that
can be selected using softkeys to move to the next display. The
text and numerical entries are only examples, and are also shown
in bold face on the displays illustrated.
This allows you to see the difference immediately between things
defined by the program and individual entries.
Explanatory text In the explanatory text, the menus and functions of the program
and the function keys are shown in pointed brackets <> and bold
face. Text and numerical inputs are placed in inverted commas
and printed in bold face. Actions that the user needs to perform
are immediately recognizable, e.g. enter ”15” and <enter>.

Preface LB 9507
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Display numbering For reasons of clarity, the displays within one measurement type
are numbered. In addition, every measurement type and special
function is given an identifying letter. This allows every display to
be easily identified and immediately assigned to a certain
procedure. The following identifying letters are used:
R = Raw data measurements (Chapter 5)
C = Cut-off measurements (Chapter 6)
L = Quantitative (LIA...) measurements (Chapter 7)
T = T-uptake/FTI measurements (Chapter 8)
D = Dual-LuciferaseTM Reporter Gene Assay (Chapter 9)
K = Kinetic measurements (Chapter 10)
W = Washing function (Chapter 11.1)
S = Call up stored data (Chapter 11.4)