MRC SPECTRO-96 Operation Manual
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MRC SPECTRO-96 Operation Manual

Fluorescent spectrophotometer
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Operation Manual for
FLUORESCENT SPECTROPHOTOMETER
SPECTRO-96
PLEASE READ THIS MANUAL CAREFULLY BEFORE OPERATION
3, Hagavish st. Israel 58817 Tel: 972 3 5595252, Fax: 972 3 5594529 mrc@mrclab.com
MRC.VER.01-5.12

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  • Page 1 Operation Manual for FLUORESCENT SPECTROPHOTOMETER SPECTRO-96 PLEASE READ THIS MANUAL CAREFULLY BEFORE OPERATION 3, Hagavish st. Israel 58817 Tel: 972 3 5595252, Fax: 972 3 5594529 mrc@mrclab.com MRC.VER.01-5.12...
  • Page 2 INSTRUCTION MANUAL FOR SPECTRO 96 FLUOROSPECTROPHOTOMETER Preface The method of fluorescence analysis (fluorometry) is a modern analytical method of high PMT level and high selectivity. It much can provide information, including an excitation spectrum, an emission spectrum, fluorescent intensity, fluorescence life, and fluorescence polarization. The linear range of working curve is wide and hence it has been an important analytical method in the field of trace analysis.
  • Page 3 INSTRUCTION MANUAL FOR SPECTRO 96 FLUOROSPECTROPHOTOMETER Alert Symbols of Safe Operation This symbol indicates important information of operating the instrument safely. Please read carefully and operate according to the instructions following. This symbol indicates a potentially hazard to the electrical appliances through the operation, and indicates that a qualified professional man is needed to deal with such a hazard according to the corresponding procedures.
  • Page 4 INSTRUCTION MANUAL FOR SPECTRO 96 FLUOROSPECTROPHOTOMETER Points for Attention to Operation 1. The Spectro 96 Fluorospectrophotometer is suitably used in a laboratory room to perform analytical measurements. If the instrument needs to be used at a worksite, the working environment of the worksite should accord with the demands of the laboratory environment basically.

  • Page 5: Table Of Contents

    INSTRUCTION MANUAL FOR SPECTRO 96 FLUOROSPECTROPHOTOMETER Contents Appearance and Performances of the Instrument ..............7 Appearance of the Complete Instrument and Main Components......7 1.1.1 The Complete Instrument .................7 Main Components ........................8 Main Components ........................9 Operating Panel of the Instrument................11 Working Principles of the Instrument................12 Signal Processing and Control System................12 Optical System ........................13 Measurement Functions of the Instrument..............14...
  • Page 6 INSTRUCTION MANUAL FOR SPECTRO 96 FLUOROSPECTROPHOTOMETER 3.4.2 Checking Wavelength of the Instrument............28 Maintenance of the Instrument....................29 Routine Maintenance ....................29 Maintenance and Replacement of Light Source Lamp..........30 4.2.1 Maintenance of Light Source Lamp ...............30 4.2.2 Replacement of Light Source Lamp ...............30 Maintenance and Replacement of Filter ..............31 4.3.1 Maintenance of Filter..................31...
  • Page 7 INSTRUCTION MANUAL FOR SPECTRO 96 FLUOROSPECTROPHOTOMETER Operation of Quantitative Analysis ................53 9.2.1 Setting of Fluorescence Wavelength for Measurement .........53 9.2.2 PMT Setting ......................53 9.2.3 Zero-point Adjustment..................53 9.2.4 Standard Sample Setting .................54 9.2.5 Measurement of Standard Samples ..............54 9.2.6 Method of Undetermined Coefficients ............55 9.2.7 Measurement of Samples.................55 9.2.8...

  • Page 8: Appearance And Performances Of The Instrument

    INSTRUCTION MANUAL FOR SPECTRO 96 FLUOROSPECTROPHOTOMETER Appearance and Performances of the Instrument Appearance of the Complete Instrument and Main Components 1.1.1 The Complete Instrument Operating Panel Heat Dissipation Opening at the Top of the Instrument Operating Panel Heat Dissipation Fan Lid of Sample Power Line Compartment...
  • Page 9 INSTRUCTION MANUAL FOR SPECTRO 96 FLUOROSPECTROPHOTOMETER Fig. 1-2 The Left Side View of SPECTRO 96 Fluorospectrophotometer Main Unit Sample Compartment (inside it a cell holder used to hold a sample cell), Power Switch (used to turn on/off the power source of the instrument), Fuse Holder (used to mount a fuse), Power Socket (used to interconnect a feed cable), Xenon Lamp Switch (used to turn on/off the Xenon lamp).

  • Page 10: Main Components

    INSTRUCTION MANUAL FOR SPECTRO 96 FLUOROSPECTROPHOTOMETER Main Components 1. Xenon Lamp Line Lens Xenon Lamp (The lightening part) Line Fig. 1-3 Xenon Lamp and Lamp Holder...
  • Page 11 INSTRUCTION MANUAL FOR SPECTRO 96 FLUOROSPECTROPHOTOMETER 2. Sample Compartment Filter Position Sample Cell Position Fig. 1-4 Sample Compartment and Quartz Sample Cell (10 mm) 3. Filter Fig. 1-5 Filter (center wavelength 365 nm)

  • Page 12: Operating Panel Of The Instrument

    INSTRUCTION MANUAL FOR SPECTRO 96 FLUOROSPECTROPHOTOMETER 1. Appearance and Performances of the Instrument 1.2 Operating Panel of the Instrument Operating Panel of SPECTRO 96 Fluorospectrop hotometer Main Unit Fig. 1-6 1. Mode key, 2. Print key, 3. Adj 0% key, 4. Normalize key, 5. Display Window.

  • Page 13: Working Principles Of The Instrument

    INSTRUCTION MANUAL FOR SPECTRO 96 FLUOROSPECTROPHOTOMETER 1. Appearance and Performances of the Instrument 1.3 Working Principles of the Instrument Signal Processing and Control System Computer Excitation Xenon Sample Cell Communication Connector Filter Lamp Controlled Wavelength Photomultiplier Display Emission Power Source of Monolithic Monochromatoraton Xenon Lamp...

  • Page 14: Optical System

    INSTRUCTION MANUAL FOR SPECTRO 96 FLUOROSPECTROPHOTOMETER Optical System Fig. 1-8 Optical System Arrangement of SPECTRO 96 Fluorospectrophotometer...

  • Page 15: Measurement Functions Of The Instrument

    INSTRUCTION MANUAL FOR SPECTRO 96 FLUOROSPECTROPHOTOMETER 1. Appearance and Performances of the Instrument Measurement Functions of the Instrument 1.4.1 Working Modes of Measurement 1. Wavelength scan mode. At a fixed excitation wavelength a spectrum is obtained in which the fluorescent intensity of a sample varies with wavelength. 2.

  • Page 16: Performance Index Of The Instrument (Specifications)

    INSTRUCTION MANUAL FOR SPECTRO 96 FLUOROSPECTROPHOTOMETER 1. Appearance and Performances of the Instrument 1.5 Performance Index of the Instrument (Specifications) Light source 150W Xenon lamp (consumable part) Excitation filter Filter character: ±5nm labeled (An interference filter with center wavelength 365nm used as standard configuration.

  • Page 17: Operation Of The Instrument

    INSTRUCTION MANUAL FOR SPECTRO 96 FLUOROSPECTROPHOTOMETER Operation of the Instrument 2.1 Operation of the Instrument Panel 2.1.1 Display Window The display window is a 128*64 pixels LCD, shows information including initialization status, fluorescence value, concentration value, emission wavelength value, PMT level, concentration factors, standard concentration, etc.

  • Page 18: Normally Working Condition Of Startup

    INSTRUCTION MANUAL FOR SPECTRO 96 FLUOROSPECTROPHOTOMETER 2. Operation of the Instrument 2.2 Normally Working Condition of Startup 2.2.1 Startup of the Instrument First turn on the power source of the xenon lamp, a Fluorospectrophotometer main unit. The xenon lamp needs to stabilize for some time when it is lit.
  • Page 19 INSTRUCTION MANUAL FOR SPECTRO 96 FLUOROSPECTROPHOTOMETER STEP PASS FAILURE Online self-checking Into on-line mode Into single mode Parameters error Database Module Database initialized Memory chip error Self- DAC Module DAC Module initialized DAC module chip error checking Photomultiplier tube error PMT Module PMT Module initialized PMT Module error...

  • Page 20: Measurement

    INSTRUCTION MANUAL FOR SPECTRO 96 FLUOROSPECTROPHOTOMETER 2. Operation of the Instrument 2.3 Measurement After preheating for 30 minutes, the instrument can be used for measurement operation. The operating methods of the two measuring modes (measuring with a single instrument, measuring with an on-line computer) of the instrument are separately different.
  • Page 21 INSTRUCTION MANUAL FOR SPECTRO 96 FLUOROSPECTROPHOTOMETER value to zero(±0.1). Attention: Zero adjustment is not allowed in the following measurement. (4) Measurement for Standard Sample. Place a series of standard solutions (from dilute to concentrated) in the light path one by one. After the fluorescence value is stable, record the value. Fit a working curve equation according to “Data Processing with Microsoft Excel 2000 Software”...

  • Page 22: Shutdown Instrument

    INSTRUCTION MANUAL FOR SPECTRO 96 FLUOROSPECTROPHOTOMETER key or the “Normalize” key to adjust the value of concentration factor. Note: When measuring the same sample, by setting the concentration factor, you may measure the sample directly. In this case, the PMT level should be in accordance with that of readout of concentration factor 2.3.3 Shutdown Instrument...

  • Page 23: Installation Of The Instrument

    INSTRUCTION MANUAL FOR SPECTRO 96 FLUOROSPECTROPHOTOMETER Installation of the Instrument 3.1 Installation Environment The instrument is suitably used in a laboratory room to perform analytical measurements. Since it works in combination with a computer, the working conditions must meet the following demands. 3.1.1 Space Environment of Laboratory Environment temperature range:10-30℃.

  • Page 24: Unpacking For Checking

    INSTRUCTION MANUAL FOR SPECTRO 96 FLUOROSPECTROPHOTOMETER 3. Installation of the Instrument Unpacking for Checking The Spectro 96 Fluorospectrophotometer main unit and the computer or the main unit and the printer are packed with a fiberboard box. For long-distance transport, users may make a request for a wooden box as additional outer packing.
  • Page 25 INSTRUCTION MANUAL FOR SPECTRO 96 FLUOROSPECTROPHOTOMETER Spectro 96 Fluorospectrophotometer main unit One piece 365nm filter (installed in the main unit) One piece Software package of data processing for One set Spectro 96 Fluorospectrophotometer (set) Power line One piece Standard USB wire One piece fittings Quartz sample cell 10mm...

  • Page 26: Installation

    INSTRUCTION MANUAL FOR SPECTRO 96 FLUOROSPECTROPHOTOMETER 3. Installation of the Instrument 3.3 Installation 3.3.1 Cleaning Remove all adhesive tapes used for fixing during transport. Clean the surface of the instrument. 3.3.2 Check of Power Source Matching Check that the power voltage to be supplied to the instrument should accord with that of the local.

  • Page 27: Adjustment Of Instrument

    INSTRUCTION MANUAL FOR SPECTRO 96 FLUOROSPECTROPHOTOMETER 3. Installation of the Instrument 3.4 Adjustment of Instrument 3.4.1 Signal Noise Ratio Test (Using Raman Spectral Peak of Water) 1..Testing Mode Using a Single Instrument Make sure the sample cell is empty. First turn on the power switch of the xenon lamp. Check that the xenon lamp is lit.
  • Page 28 INSTRUCTION MANUAL FOR SPECTRO 96 FLUOROSPECTROPHOTOMETER 2..Testing Mode by using the Instrument in Combination with an On-line Computer Finish the measurement for Raman Spectral Peak of water by using wavelength scan and time scan in the computer software. The theory is the same as Section 3.4.1 Part 1. The use of software please turn to Part Two.

  • Page 29: Checking Wavelength Of The Instrument

    INSTRUCTION MANUAL FOR SPECTRO 96 FLUOROSPECTROPHOTOMETER 3. Installation of the Instrument 3.4.2 Checking Wavelength of the Instrument The instrument can automatically check the accuracy of indicated value of the instrument wavelength. If finding a mistake, may use a mercury-arc lamp to check (If there is no mercury-arc lamp, a fluorescent lamp can be used instead, but a professional man must be needed to do such work.

  • Page 30: Maintenance Of The Instrument

    INSTRUCTION MANUAL FOR SPECTRO 96 FLUOROSPECTROPHOTOMETER Maintenance of the Instrument Routine Maintenance 1. In routine operation, always check and make sure that the working conditions accord with the demands. If finding that the working conditions do not accord with the demands, should take measures without delay.

  • Page 31: Maintenance And Replacement Of Light Source Lamp

    INSTRUCTION MANUAL FOR SPECTRO 96 FLUOROSPECTROPHOTOMETER 4. Maintenance of the Instrument Maintenance and Replacement of Light Source Lamp 4.2.1 Maintenance of Light Source Lamp 1.A high-performance xenon lamp is used as the light source lamp of the instrument. It is mounted in a special lamp chamber as shown in Fig.1-3.

  • Page 32: Maintenance And Replacement Of Filter

    INSTRUCTION MANUAL FOR SPECTRO 96 FLUOROSPECTROPHOTOMETER 4. Maintenance of the Instrument 4.3 Maintenance and Replacement of Filter 4.3.1 Maintenance of Filter 1. The surface of the filter should be kept clean. Note: If having touched the surface of the filter, please wipe it clean with a piece of lens tissue and alcohol then dry it.
  • Page 33 INSTRUCTION MANUAL FOR SPECTRO 96 FLUOROSPECTROPHOTOMETER Part Two: Operation Instruction for the Instrument Connected with a Computer...

  • Page 34: Environmental Requirements Of Using Software And Installation

    INSTRUCTION MANUAL FOR SPECTRO 96 FLUOROSPECTROPHOTOMETER Environmental Requirements of Using Software and Installation Before reading this part, please first read the “Operation Instruction for the Instrument” in Part Two and the operation manuals for the above versions of Windows 98. Basic Configuration of Computer Hardware and Software 5.1.1 Hardware CPU P2 Celeron is 600 or more.

  • Page 35: Installation Of "F96 Software

    INSTRUCTION MANUAL FOR SPECTRO 96 FLUOROSPECTROPHOTOMETER 5．Environmental Requirements of Using Software and Installation 5.2 Installation of “Spectro 96 Software” Place CD-ROM into the drive. The computer will automatically install an application. In the absence of automatic installation, open the X: \ (double-click) SETUP.EXE documents (applications).

  • Page 36: Operation Of Software

    INSTRUCTION MANUAL FOR SPECTRO 96 FLUOROSPECTROPHOTOMETER Operation of Software 6.1 Preparation before Use 6.1.1 Connection of Spectro 96 Main Unit with a Computer The Spetro96 main unit is connected through a USB por t of a computer. The USB driver will be installed when the first time connected.
  • Page 37 INSTRUCTION MANUAL FOR SPECTRO 96 FLUOROSPECTROPHOTOMETER EM Wavelength Storage Fluor. Fluor. Fig. 2-1 Interface of Wavelength Scan Wavelength Scan Fig. 2-2 Switchover of Working Mode...

  • Page 38: Three Interfaces Of Working Mode

    INSTRUCTION MANUAL FOR SPECTO 96 FLUOROSPECTROPHOTOMETER 6. Operation of Software 6.2 Three Interfaces of Working Mode There are Three Interfaces of Working Mode, namely the interface of wavelength scan, the interface of time scan and the interface of quantitative analysis. 6.2.1 The Interface of Wavelength Scan Wavelength scan means that in the conditions of selected excitation source (namely at excitation wavelength) people measure the fluorescent intensity which varies with fluorescent...

  • Page 39: The Interface Of Quantitative Analysis

    INSTRUCTION MANUAL FOR SPECTRO 96 FLUOROSPECTROPHOTOMETER Time scan differs from wavelength scan in that the unit of abscissa is time during scanning. In the functions such as “Set Parameter”, “Spectrum”, “File” and operation, the requirements for time scan are the same as those for wavelength scan. 6.2.3 The Interface of Quantitative Analysis Quantitative analysis means that in the conditions of selected excitation source (namely at excitation wavelength) by measuring the fluorescent emission intensity (F) of the component (its...

  • Page 40: Operation Of Wavelength Scan Mode (Connected With A Computer)

    INSTRUCTION MANUAL FOR SPECTRO 96 FLUOROSPECTROPHOTOMETER Operation of Wavelength Scan Mode (Connected with a Computer) The Interface of Wavelength Scan Mode On completion of initialization for the instrument, the interface of wavelength shown in Fig.2-5 is immediately displayed. The first row at the top of screen is Menu Bar. In the second row are 15 “Shortcut Icons”.

  • Page 41: The Functions Of Shortcut Icons

    INSTRUCTION MANUAL FOR SPECTRO 96 FLUOROSPECTROPHOTOMETER “Spectrum Parameter” is the button to show the parameters of current wavelength scan. 7.1.2 The Functions of Shortcut Icons When the mouse pointer is moved onto a shortcut icon, the function of the shortcut icon will show.

  • Page 42: Wavelength Scanning Operation

    INSTRUCTION MANUAL FOR SPECTRO 96 FLUOROSPECTROPHOTOMETER 7. Operation of Wavelength Scan Mode (When Connected with a Computer) 7.2 Wavelength Scanning Operation Fluorescence spectrum can be acquired by wavelength scan, and the wavelength of fluorescent measurement used for quantitative analysis can be determined from the fluorescence spectrum.
  • Page 43 INSTRUCTION MANUAL FOR SPECTRO 96 FLUOROSPECTROPHOTOMETER 8.PMT Level. Set the PMT level of time scan. 9.Multi-Curve. Set the display mode of curves. “Overlap” shows different curves in the same window. “Overlay” will cover the last time scan curve, only showing the latest. 10.

  • Page 44: Spectrum Processing

    INSTRUCTION MANUAL FOR SPECTRO 96 FLUOROSPECTROPHOTOMETER Fig. 2-8 Interface of Wavelength Scan Parameter – Abbreviatory Curve 15.Pre-Scan. It’s a fast scan according to “Basic Settings” in “Wavelength Scan Parameter”. After a Pre-Scan, a thumbnail is created and proper Y axis will be automatically set. 7.2.2 Spectrum Processing Clicking the “...
  • Page 45 INSTRUCTION MANUAL FOR SPECTRO 96 FLUOROSPECTROPHOTOMETER spectrum to make the spectrum smoother. 5. P/ V Detection. It is used to detect the values of peaks and valleys of a spectrum. (1) Click this item and “Input Sensitivity” dialog box will appear. Input a limited value of “Input Sensitivity”...

  • Page 46: Save Processing

    INSTRUCTION MANUAL FOR SPECTRO 96 FLUOROSPECTROPHOTOMETER 7.2.3 Save Processing Click “File” in Fig.2-5 and a pull-down sub-menu is shown in Fig.2-11. Fig. 2-11 Save Processing Sub-menu 1. Select Channel. 10 channels (storage windows) altogether are provided for selection. Program can separately open 10 storage windows of wavelength scan or time scan at most for users’ use.

  • Page 47: Display Setting

    INSTRUCTION MANUAL FOR SPECTRO 96 FLUOROSPECTROPHOTOMETER 7.2.4 Display Setting Click “View” in Fig.2-5. A pull-down sub-menu is shown in Fig.2-12. 1. Line Color. Change the color of the current spectrum. 2. Back Color. Change the background color of the working window. 3.

  • Page 48: Operation Of Time Scanning Mode (When Connected With A Computer)

    INSTRUCTION MANUAL FOR SPECTRO 96 FLUOROSPECTROPHOTOMETER 8 Operation of Time Scanning Mode (When Connected with a Computer) Interface and Operation of Time Scanning Mode Time scan differs from wavelength scan in that the unit of X-axis is time during scanning. This working mode can be used to observe the situation in which the fluorescent intensity of a sample varies with time at a fixed emission wavelength.

  • Page 49: Spectrum Processing

    INSTRUCTION MANUAL FOR SPECTRO 96 FLUOROSPECTROPHOTOMETER 6. Replicates. Set the number of scans. 7. Interval. Set the update interval of time scan. Update every 0.1 second. 8.PMT. Set the signal PMT level of time scan. 9.Most Data. Set the maximum sample size of time scan. 10.Multi-Curve.

  • Page 50: Operation Of Quantitative Analysis Mode (When Connected With A Computer)

    INSTRUCTION MANUAL FOR SPECTRO 96 FLUOROSPECTROPHOTOMETER Operation of Quantitative Analysis Mode (When connected with a Computer) Interface of Quantitative Analysis Mode Click “Working Mode” on the upper left screen. The interface of quantitative analysis mode from the pull-down menu will appear as shown in Fig.2-15 Fig.

  • Page 51: The Functions Of Short Cuts On The Bottom Of The Screen

    INSTRUCTION MANUAL FOR SPECTRO 96 FLUOROSPECTROPHOTOMETER 9.1.1 The Functions of Short cuts on the Bottom of the Screen The “Set Standard” button is used to set standard sample. After clicking it, a “Set Standard Sample” dialog will appear. For more details, see Fig. 2-20. The “Measure Sample”...

  • Page 52: Menu On The Top Of The Screen

    INSTRUCTION MANUAL FOR SPECTRO 96 FLUOROSPECTROPHOTOMETER 9.1.3 Menu on the Top of the Screen Menu on the top of the screen has columns such as “Analysis Mode”, “Set Parameter”, “Curve Type”, “Spectrum”, “Data”. 1. Analysis Mode This feature has not been activated yet. 2.
  • Page 53 INSTRUCTION MANUAL FOR SPECTRO 96 FLUOROSPECTROPHOTOMETER ⑶ ⑶ ⑶ ⑶ Save Curve As. Save Curve As. Save Curve As. Save and rename the current curve. Save Curve As. ⑷ ⑷ ⑷ ⑷ Delete Single Spl. Data. Delete the data of a single sample from the sample column. ⑸...

  • Page 54: Operation Of Quantitative Analysis

    INSTRUCTION MANUAL FOR SPECTRO 96 FLUOROSPECTROPHOTOMETER 9. Operation of Quantitative Analysis Mode (When connected with a Computer) Operation of Quantitative Analysis The interface of quantitative analysis is shown in Fig. 2-19. On the left is the display box of Standard Sample and on the right is the display box of Unkown Sample. 9.2.1 Setting of Fluorescence Wavelength for Measurement Click “Set Standard”...

  • Page 55: Standard Sample Setting

    INSTRUCTION MANUAL FOR SPECTRO 96 FLUOROSPECTROPHOTOMETER 9.2.4 Standard Sample Setting After clicking the “Set Standard.” shortcut on the bottom left of the interface of quantitative analysis, a dialog box of “Set Standard Samples” shown in Fig.2-18 appears. Concentration unit can be set in “Unit” box. If there are 5 standard samples and their concentrations are 10 for No.1 sample, 20 for No.2 sample, 30 for No.3…….respectively.

  • Page 56: Method Of Undetermined Coefficients

    INSTRUCTION MANUAL FOR SPECTRO 96 FLUOROSPECTROPHOTOMETER 9.2.6 Method of Undetermined Coefficients Click “Curve Type” in the menu bar. Select the type of curve that you need in the pull-down sub-menu (The default setting is linear curve.). Then click “Data” in the menu bar and select “Build Curve” in the sub-menu. A “Working Curve Window”...

  • Page 57: Appendix: Data Processing With Microsoft Excel 2000

    INSTRUCTION MANUAL FOR SPECTRO 96 FLUOROSPECTROPHOTOMETER Appendix: Data Processing with Microsoft Excel 2000 Data Processing with Microsoft Excel 2000 Software Experimental data processing of fluorophotometry by using Excel 2000 Software. Open Microsoft Excel 2000 software. The interface of the software will be displayed on the screen of the computer.
  • Page 58 INSTRUCTION MANUAL FOR SPECTRO 96 FLUOROSPECTROPHOTOMETER Legend”. (3) Insert Graphics. Click the “Next” button to go into “Chart Wizard 4-Procedure 4”. Select the location of inserting a graphics. 3. Data Processing by using Linear Regression From the Excel menu select “Chart (C)”→“Add Trend Line (R)…”. Select “Linear Regression”.

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