MRC SPECTRO-96 User manual

PLEASE READ THIS MANUAL CAREFULLY BEFORE OPERATION
3, Hagavish st. Israel 58817 Tel: [phone], Fax: [phone] [email protected]
MRC.VER.01-5.12
Operation Manual for
FLUORESCENT SPECTROPHOTOMETER
SPECTRO-96

1
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.
The Spectro 96 Fluorospectrophotometer is a fluorophotometer using filters
with a monochromator. It has three functions, namely, wavelength scan, time
scan for fluorescent intensity, and quantitative analysis. The instrument is
capable of performing two modes of measurement: measuring with a single
instrument only, and measuring with an on-line computer. The amounts of the
functions and the convenience of measuring vary with the different modes of
measurement.
For proper use of the Spectro 96 Fluorospectrophotometer, the user of the
instrument should have a basic knowledge of using optical instruments and a
basic knowledge of molecular fluorescence spectroscopy. The user whose
instrument is equipped with a computer also should have a corresponding
knowledge and skills of operating the computer.
Before installation or the first use of this instrument, the user must read
through this instruction manual carefully. According to the user’s different
equipment, the spectrophotometer main unit may be equipped with an external
computer or a special printer.
Our company guarantees that if the instrument is subject to damage or
fails to work normally due to the imperfect manufacture of our product, the
company will be responsible for the repair of the instrument (excluding
quick-wear parts and consumables) without charge within the warranty period of
12 months from the delivery date under the condition that the user complies with
the rules of transport, maintenance and operation. If the instrument is in need of
repair, please send it and its accessories back to our company, along with the
quality assurance card of our product.
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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. (This label is attached on
the power switch and trigger.)
This symbol indicates heating on the surface. Please
perform cautiously according to the instructions
following.
!
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Points for Attention to Operation
1. The Spectro 96 Fluorospectrophotometer is suitably usedin 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.
2. If the user needs to move the instrument to other working site for analytical measurements,
please use original packing materials on moving.
3. Be sure to turn on⁄off the power of the instrument according to the operating procedure.
When starting up the instrument, please turn on the power source of the xenon lamp first.
Then turn on the power source of the spectrophotometer main unit after the xenon lamp is
lit. When shutting down the instrument, please turn off the power source of the
spectrophotometer first and then the power source of the xenon lamp. Otherwise, the
instrument may not work normally. After the xenon lamp is lit, it is necessary to wait for 30
minutes over till the lamplight becomes stable. After the power source of the xenon lamp is
turned off, please wait for 60 seconds, and then turn on the lamp if the lamp needs to be
used again.
4. When the power source of the xenon lamp is turned on but the lamp fails to be lit and a
squeaking sound of high frequency or a cracking sound of sparking is produced
continuously, please turn off the power source of xenon lamp immediately and wait for
several seconds, then turn on the lamp again. Because the life of the xenon lamp is closely
related to the number of on-off of the lamp power source, the lamp should avoid being
turned on/off repeatedly under a high voltage and the unnecessary number of on/off of the
xenon lamp power source should be cut down as far as possible.
5. When the instrument is working, the surface temperature of the heat dissipation opening at
the top of the left back side of the instrument (xenon lamp position) becomes very high. Be
sure not to touch the surface of the heat dissipation opening with hands in order not to be
burnt. The air around the heat dissipation opening should be kept circulating.
6. After each startup of the instrument, please first confirm that the heat dissipation fan at the
side of the instrument runs normally in order to ensure normal work of the instrument. If
the user finds anything wrong with the heat dissipation fan, the instrument should be closed
down for an overhaul.
7. In order to protect the photomultiplier when PMT level is higher (>6), do not let high light
pass into the sample compartment. When a sample of unknown concentration is measured,
PMT level should be set from low grade to high grade (1-17) gradually.
8. After PMT level is adjusted, the original set for zero-point of fluorescence should be
checked and note that zero-point adjustment may be needed again.
9. When and only when a computer error is due to the operator¹s false operation or other
interferences, the power switch of the spectrophotometer main unit should be turned off
immediately. When the software cannot be operated, please use task manager to end the
“f96pc.exe” process, then restart the software and the instrument.
10. Within the monochromator the tightened screws must not be loosened. The optical devices
and the working environment of the instrument should be kept clean.
11. When opening the cover to inspect the instrument, be sure to turn off the power source.
Particularly after the upper cover of the instrument is removed, as there are electrical
appliance parts with high voltage located at the left back of the instrument base, be sure not
to contact these parts while the power source is still turned on.
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Contents
1Appearance and Performances of the Instrument..................................................................7
1.1 Appearance of the Complete Instrument and Main Components.............................7
1.1.1 The Complete Instrument .................................................................................7
Main Components................................................................................................................8
Main Components................................................................................................................9
1.2 Operating Panel of the Instrument.............................................................................11
1.3 Working Principles of the Instrument........................................................................12
Signal Processing and Control System...............................................................................12
Optical System...................................................................................................................13
1.4 Measurement Functions of the Instrument................................................................14
1.4.1 Working Modes of Measurement....................................................................14
1.4.2 Modes of Data Measurement...........................................................................14
1.4.3 Self-checking and Self-correcting ...................................................................14
1.5 Performance Index of the Instrument (Specifications).............................................15
2Operation of the Instrument...................................................................................................16
2.1 Operation of the Instrument Panel.............................................................................16
2.1.1 Display Window................................................................................................16
2.1.2 Operation of the Function Keys......................................................................16
2.2 Normally Working Condition of Startup...................................................................17
2.2.1 Startup of the Instrument................................................................................17
2.2.2 Confirmation of Heat Dissipation Fan Running............................................17
2.2.3 Initialization......................................................................................................17
Table 1-2 Fluorospectrophotometer Initialization List.........................................18
2.3 Measurement................................................................................................................19
2.3.1 Pouring the Sample Solution into Sample Cell..............................................19
2.3.2 QuantitativeAnalysis in Measurement Mode with a Single Instrument.....19
2.3.3 Shutdown Instrument......................................................................................21
3Installation of the Instrument.................................................................................................22
3.1 Installation Environment.............................................................................................22
3.1.1 Space Environment of Laboratory.................................................................22
3.1.2 Working Table ..................................................................................................22
3.1.3 Power Source ....................................................................................................22
3.1.4 Environmental Alternation..............................................................................22
3.2 Unpacking for Checking..............................................................................................23
3.2.1 Checking the Outer Packing ...........................................................................23
3.2.2 Unpacking for Counting..................................................................................23
3.3 Installation....................................................................................................................25
3.3.1 Cleaning ............................................................................................................25
3.3.2 Check of Power Source Matching...................................................................25
3.3.3 Connecting the Power Cord ............................................................................25
3.4 Adjustment of Instrument...........................................................................................26
3.4.1 Signal Noise Ratio Test (Using Raman Spectral Peak of Water).................26
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3.4.2 Checking Wavelength of the Instrument........................................................28
4Maintenance of the Instrument...............................................................................................29
4.1
4.14.1
4.1 Routine Maintenance...................................................................................................29
4.2
4.24.2
4.2 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
4.3 Maintenance and Replacement of Filter....................................................................31
4.3.1 Maintenance of Filter.......................................................................................31
4.3.2 Replacement of Filter.......................................................................................31
5Environmental Requirements of Using Software and Installation......................................33
5.1 Basic Configuration of Computer Hardware and Software.....................................33
5.1.1 Hardware..........................................................................................................33
5.1.2 Software Requirements....................................................................................33
5.2 Installation of “F96 Software” ....................................................................................34
6Operation of Software..............................................................................................................35
6.1 Preparation before Use................................................................................................35
6.1.1 Connection of Spectro96Main Unit with a Computer......................................35
6.1.2 Procedure for On-line Operation (Startup)...................................................35
6.2 Three Interfaces of Working Mode.............................................................................37
6.2.1 The Interface of Wavelength Scan..................................................................37
6.2.2 The Interface of Time Scan.............................................................................37
6.2.3 The Interface of Quantitative Analysis...........................................................38
7Operation of Wavelength Scan Mode (Connected with a Computer).................................39
7.1 The Interface of Wavelength Scan Mode...................................................................39
7.1.1 The Functions of Shortcuts at the Bottom of the Screen..............................39
7.1.2 The Functions of Shortcut Icons.....................................................................40
7.1.3 Menu Bar at the Top of the Screen.................................................................40
7.2 Wavelength Scanning Operation ................................................................................41
7.2.1 Parameter Setting.............................................................................................41
7.2.2 Spectrum Processing........................................................................................43
7.2.3 Save Processing.................................................................................................45
7.2.4 Display Setting..................................................................................................46
8Operation of Time Scanning Mode (When Connected with a Computer)..........................47
8.1 Interface and Operation of Time Scanning Mode.....................................................47
8.1.1 Interface of Time Scanning Mode...................................................................47
8.1.2 Parameter Setting.............................................................................................47
8.1.3 Spectrum Processing........................................................................................48
8.1.4 Storage Processing............................................................................................48
8.1.5 Display Setting..................................................................................................48
9Operation of QuantitativeAnalysis Mode (When connected with a Computer)................49
9.1 Interface of Quantitative Analysis Mode ...................................................................49
9.1.1 The Functions of Short cuts on the Bottom of the Screen ............................50
9.1.2 The Functions of Shortcuts of Icon.................................................................50
9.1.3 Menu on the Top of the Screen........................................................................51
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9.2 Operation of QuantitativeAnalysis............................................................................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 Set Sample Name..............................................................................................55
10 Appendix: Data Processing with Microsoft Excel 2000................................................56
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1Appearance and Performances of the Instrument
1.1 Appearance of the Complete Instrument and Main Components
1.1.1 The Complete Instrument
Operating Panel
Fig. 1-1 SPECTRO 96 Fluorospectrophotometer Main Unit
Heat Dissipation Opening at the Top of the Instrument
Heat Dissipation Fan
Power Line Lid of Sample
Compartment
Operating Panel
USB Interface
RS232 Communication Connector
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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).
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Main Components
1. Xenon Lamp
Fig. 1-3 Xenon Lamp and Lamp Holder
Line
Xenon Lamp (The
lightening part)
Line
Lens
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Fig. 1-5 Filter (center wavelength 365 nm)
2. Sample Compartment
Fig. 1-4 Sample Compartment and Quartz Sample Cell (10 mm)
3. Filter
Filter Position
Sample Cell
Position
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Fig. 1-6 Operating Panel of SPECTRO 96 Fluorospectrop hotometer Main Unit
1. Mode key, 2. Print key, 3. Adj 0% key, 4. Normalize key, 5. Display Window.
2
5
4
3
1
1.2 Operating Panel of the Instrument
1.
Appearance and Performances of the Instrument
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1.3 Working Principles of the Instrument
Signal Processing and Control System
Fig. 1-7 Signal Processing and Control System
1.
Appearance and Performances of the Instrument
Xenon
Lamp
Power Source of
Xenon Lamp
Excitation
Filter Sample Cell
Emission
Monochromatoraton
Photomultiplier
Preamplifier
Monolithic
Processor
Serial Interface
USB
Communication Connector
Computer
Serial Printer
Negativ e
High Voltage
Display
Key
Board
Controlled
Wavelength
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Optical System
Fig. 1-8 Optical System Arrangement of SPECTRO 96 Fluorospectrophotometer
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1.4 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. Time scan mode. At a fixed excitation wavelength, a spectrum is obtained in which the
fluorescent intensity of a sample varies with time.
3.Quantitative analysis mode. According to the quantitative relationship of fluorophotometry
F=KC, under a given experimental condition, fluorescent intensity (F) is directly proportional to the
concentration (C) of a sample component to be determined. By measuring the fluorescent intensity
of a standard solution, K is acquired by curve fitting. Then measure the fluorescent intensity of an
unknown sample. From a quasi-linear equation calculate the concentration of the component to be
determined in the unknown sample. A calibration can also be made by standard curve method or
standard addition method.
A concentration measurement can be made through direct reading from the instrument by
single-point standard calibrating method.
1.4.2 Modes of Data Measurement
1. Measuring with a single instrument. By using the functions of the keys and the functional
indicator lights on the instrument panel, read the measured data on the display window.
2. Measuring with an on-line computer. The instrument is connected with a computer through a
communication connector. The instrument control, the parameters setting, and real-time
measurement are all carried out by a computer software system. The software possesses powerful
functions of spectrum processing, data processing and storage.
Note: For different modes of data measurement, the number of usable working modes and the
convenience of use differ somewhat.
1.4.3 Self-checking and Self-correcting
1. Self-checking and self- correcting function of the instrument
The instrument will automatically initialize after startup, including self-checking and
wavelength correcting. Self-checking contains on-line condition check, excitation source check,
database module check, DAC module check, PMT module check and motor module check. The
result of each check will be printed on the LCD or in the software window when connected to
computer. Users can check out problems refer to table 1-2. Wavelength correcting program will
guarantee the accuracy of wavelength.
1.
Appearance and Performances of the Instrument
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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. Users may separately purchase other φ25mm
interference filters with center wavelength within 200―850nm range.)
Emission
monochromator
Emission monochromator wavelength range: (Em) 200―900nm
Wavelength accuracy within ±1nm
Wavelength repetition ≤0.5nm
Emission spectrum bandwidth: 10nm
S⁄N ratio S/N≥150 (Using 1cm quartz sample cell, measure the signal noise ratio
of Raman spectrum of water )
Check utmost 1×10
-10
g/ml sulfuric acid quinine solution
Measure linearity ≥0.995
Peak value ≤1.5%
Zero line drift ≤0.3% (Within 10 minutes)
Upper limit change
of indicating value
±1.5% (Within 10 minutes)
Power requirement 220V±22V 110V/60HZ
Dimensions 442×392×250（mm）
Net weight 11kgWeight
Gross weight 13kg
Table 1-1 Performance Index of the Instrument
1.
Appearance and Performances of the Instrument
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2Operation 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.
Symbol related:
1. “ ●
●●
● ” means the excitation light source is on.
2. “%” means the fluorescence value has been normalized.
2.1.2 Operation of the Function Keys
1. “Mode” Key : Select the operation mode. The switch circulation is as follow: Fluorescence
Value Mode -> Wavelength Mode -> PMT Level Adjust -> Concentration Input Mode -> Factors
Input Mode -> Concentration Mode -> Fluorescence Value Mode.
In Wavelength Mode and PMT LevelAdjust, if parameter value set is different from current
value, use “Mode” key to cancel current value and return the set value.
2. “Print” Key: The “Print” key can print the current value if a dedicated serial interface printer
is installed in Fluorescence Value Mode and Concentration Mode. In other modes, the “Print” key
confirm the changed parameters.
3. “Adj 0” Key: In Fluorescence Value Mode, The key can adjust the current value to zero. In
Wavelength Mode, Concentration Input Mode and Factors Input Mode, the key can select every
single number from left to right. While selected, the value can be changed. In PMT Level Adjust,
The key can reduce PMT level.
4. “Normalize” Key: In Fluorescence Value Mode, The key will normalize current value to 100
and displays “%” symbol. Press again will cancel the normalization. In Wavelength Mode,
Concentration Input Mode and Factors Input Mode, the key will add the selected number from 0-9
in circulation. In PMT Level Adjust, The key can add PMT level.
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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. For precise measurement, the time should be 30 minutes over.
Note :
① If the xenon lamp fails to be lit and a squeaking sound of high frequency or a cracking
sound of sparking is continuously produced, please turn off the power source of xenon lamp
immediately. Several seconds later, then turn on the xenon lamp again.(Such a case occurs only
when the power source is not stable and capacity of the power source is not sufficient, or xenon
lamp closes to the limit of service life).
②Because the life of the xenon lamp is closely related to the number of on-off of the lamp
power source, the unnecessary number of turning on/off the xenon lamp should be cut down as far
as possible and the xenon lamp should avoid being turning on/off repeatedly under a high voltage.
2.2.2 Confirmation of Heat Dissipation Fan Running
Every time after startup of the instrument, please first make sure that the heat dissipation fan at
the side of the instrument runs normally in order to ensure that the instrument works normally. If
finding anything wrong with the heat dissipation fan, should close down the instrument for an
overhaul.
2.2.3 Initialization
The instrument will automatically initialize after startup. If the instrument is not connected to
computer, the initialization status will be displayed on LCD. If connected to computer, the status
will be displayed in software window, and the LCD will show “On-line”.
Initialization includes self-checking and self-correcting. Information of every step is given in
table 1-2.
2.
Operation of the Instrument
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STEP PASS FAILURE
Online self-
checking
Into on-line mode Into single mode
Database Module Database initialized Parameters error
Memory chip error
DAC Module DAC Module initialized DAC module chip error
PMT Module PMT Module initialized
Photomultiplier tube error
PMT Module error
Self-
checking
Motor Module Motor Module initialized Motor error
Motor driver board error
Self-
correcting
Wavelength
correcting
T
he value from bottom to
top and to bottom again,
shows a wave peak.
No obvious wave peak.
Wavelength error
Check if the sample cell is
empty
Table 1-2 SPECTRO 96 Fluorospectrophotometer Initialization List
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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.
When the Fluorospectrophotometer main unit is connected with a computer, the
selection of the different measuring modes and the operating methods are seen in detail in Section 7,
Section 8 and Section 9 of Part Two.
This section is about the operation of quantitative analysis using a single instrument for
measurement.
2.3.1 Pouring the Sample Solution into Sample Cell
Pour the sample solution to be measured into the sample cell up to about two thirds of height.
Use filter paper to remove the solution left on the outside surface of the sample cell. Place it
carefully into the sample cell holder.
Note: The sample cell is made of quartz material. When using, should take care to protect the
window of light transmission. When taking the sample cell out or placing it into the holder with
hand, should hold the sample cell diagonally with fingers.
2.3.2 Quantitative Analysis in Measurement Mode with a Single Instrument
Measuring mode with a single instrument is mainly used in the working mode for quantitative
analysis. There are two methods, that is multipoint method of standard sample and single-point
method of standard sample. The two operating methods are as follows.
1. Measuring procedures by Multipoint Method of Standard Sample
Prepare a series of standard solutions (including blank solution).After finishing the preparation
of the solution of unknown sample, operate as the following sequence.
(1) Wavelength Adjustment. Press the “Mode” key to enter Wavelength Mode. “Current
Wavelength: ***nm” shows current wavelength of emission monochromator. Use “Adj 0” key or
the “Normalize” key to adjust target value. Press “Print” key after adjustment(See 2.1.2 for detail).
The wavelength will be automatically set to target value.
(2) PMT LevelAdjustment. Press the “Mode” key into Fluorescence Value Mode. Put the
standard solution of maximal concentration into sample cell. Then press “Mode” to enter PMT level
adjust. “Current PMT level: **” shows the current PMT level. The higher the level is, the more
sensitive it will be. Use “Adj 0” key or the “Normalize” key to adjust the PMT level. Press “Print”
key after adjustment. The PMT level will be automatically set to target value. Adjust the PMT level
to make sure the fluorescence value of the maximal concentration standard solution is set between
200-400.
(3) Zero Adjustment. Place the blank solution in the light path. Press the “Mode” key into
Fluorescence Value Mode. Wait till the value is stable. Press “Adj 0” to adjust the fluorescence
2.
Operation of the Instrument
INSTRUCTION MANUAL FOR SPECTRO 96 FLUOROSPECTROPHOTOMETER