MRC Spectro-97 Operation Manual

Fluorescent Spectrophotometers
SPECTRO-97 SERIES
Operation Manual
PLEAS
E READ THIS MANUAL CAREFULLY BEFORE OPERATION
Hagavish st. Israel 58817
Tel: 972 3 5595252, Fax: 972 3 5594529 mrc@mrclab.com
MRC.4.18
1
Preface
Thank you for purchasing the Fluorospectro­photometer of MRC Ltd. Please read this manual carefully before installation or first time using SPECTRO-97 series Fluorospectrophotometer. Fluorometry i
s a high sensitive, high selective and modern analysis method. Fluorometry provides excitation spectrum, emission spectrum, including luminous intensity, luminous life, fluorescence polarization and other information. It’s linear range of working curve is wide enough to become an important analysis method in Measure analysis. For proper use of SPECTRO-97 series Fluorospectrophtometer, basic knowledge of optical instruments and Molecular Fluorometry is needed. Computer operation skills are needed as well.
SPECTRO-97 series includes SPECTRO-97, SPECTRO-97 Pro and SPECTRO-97XP 3 kinds of
Fluorospectrophtometers. For more detail of parameter differences, please check manual chap.
1.4.
SPECTRO-97 series Fluorospectrophtometer
is a dual-monochromator fluorospec
trophtometer with fluorescence excitation wavelength scan, emission wavelength scan, 3D scan, Synchronous scan, time scan, quantitative analysis and other functions. The Fluorospectrophtometer should be operated with computer. This manual includes instrument instruction, software instruction and appendix. Chap. 6.3.5 is a value-added part. If you need this part, please contact us.
2
Special Statement
Pleas
e read this manual carefully before installation or operation. The company will not take responsible for any trouble or damage due to unproper use. The company has the final interpretation of this manual. Modifications of the manual due to improvements of the instrument will not be announced. The company will conduct 12 months free repair from the date of delivery if the instrument is in strict accordance withthe instructions and the transport safety specification. (Vulnerability and consumable parts are not included) Please useouroriginal packaging when returning the instrumentfor service with accessories and the warranty card. This manual is for SPECTRO-97series Fluorospectrophtometer. The company will post updates of all the information on our website.
Check out our website for more information. For more service, please dial our marketing department 972-3-5595252, or fax 972-3-5594529. Any chapter or images of this manual are not allowed to borrow, copy and translate to other languages without permission of the company.
3
Safety Signs
This sign indicates important part of the
instrument. Please follow instructions.
Please read and operate according to
the instructions following.
This sign indicates possible electric
harm.
Required by a professionally
qualified personnel according to the
appropriate procedures. (This label is
attached on the power switch and
trigger.)
This sign indicates heat o
n surface.
!
4
Precautions
1. The instrument is suitable for analysis in laboratory. If
the instrument is needed outside the lab, please make the field work environment meets the environmental requirements of the laboratory.
2. Please use our original packaging when moving
the
instrument.
3.
Please boot the instrument in sequence. First, t
urn
on
the Xenon lamp power supply. Then turn on the main power. When shutting down, turn off the main power first, then turn off the Xenon lamp power supply. There will be 30 min before Xenon lamp becomes steady. Please wait 60 sec to retrigger the Xenon lamp if the Xenon lamp power shut down.
4. If the Xenon lamp power is not triggered and making noise, shut down the Xenon lamp power immediately and retrigger after 60 sec. Due to the fact that Xenon lamp life is closely related to the switching times, please minimize unnecessary Xenon lamp trigger times.
5. When the instrument is on, the temperature of th
e
vents
on top left corner is high. Please keep the air circulating and away from the vents surface. DO NOT observed Xenon light directly with naked eye.
6. Please make sure the fans on the left side and t
op
left
corner operate normally. If the fans are not functioning, please turn off the instrument for repairs.
7. As to protect PMT, DO NOT let light into the
sample
cell
when the gain is higher than 6. When using unknown sample in the test, set the gain from low to high gradually from 1 to 17.
8. Please check the Fluorescent zero and adjust
zero
after setting gain.
9.
When an error occurred by wrong operation or o
ther
5
machine or instrument error, shut down the instrument immediately. When the software is not operating properly, Start Task Manager to end the "NeoLG.exe" process, then restart the software and the instrument.
10. DO NOT loose the screws in the monochromator. Keep the environment clean.
11.Cut the power before opening the instrument. Pay attention to the high-voltage electrical components on the left rear of the instrument.
12. Cover the instrument with dustproof if
the
instrument is not used for a long time.
6
Menu
Preface ............................................................................................................................................... 1
Special Statement .............................................................................................................................. 2
Safety Signs ...................................................................................................................................... 3
Precautions ........................................................................................................................................ 4
PART IU s e r ’ s G u i d e ........................................................................ 9
1 Appearance & Performance ......................................................................................................... 10
1.1 Appearance ........................................................................................................................ 10
1.1.1 Body ....................................................................................................................... 10
1.1.2 Interface ................................................................................................................. 11
1.1.3 Power & Switches .................................................................................................. 12
1.1.4 Sample Compartment ............................................................................................. 13
1.2 Mode of Operation ............................................................................................................ 14
1.2.1 Signal Processing & Control System ..................................................................... 14
1.2.2 Light Path ............................................................................................................... 15
1.3 Functions ........................................................................................................................... 15
1.3.1 Modes For Measurement ........................................................................................ 16
1.3.2 Self Tests & Adjustments ....................................................................................... 16
1.4Performance ....................................................................................................................... 16
2 Booting & Shutting Down ........................................................................................................... 18
2.1 Booting Status ................................................................................................................... 18
2.1.1 Booting ................................................................................................................... 18
2.1.2 Fans Condition ....................................................................................................... 18
2.1.3 Multi-instruments Booting ..................................................................................... 19
2.1.4 Initialization ........................................................................................................... 19
2.2 Power Off .......................................................................................................................... 19
3 Installation .................................................................................................................................... 20
3.1Environment ....................................................................................................................... 20
3.1.1 Laboratory Environment ........................................................................................ 20
3.1.2 Work Bench ............................................................................................................ 20
3.1.3 Power ..................................................................................................................... 20
3.1.4Environment Change ............................................................................................... 20
3.2 Package ............................................................................................................................. 20
3.2.1 Check the Package ................................................................................................. 20
3.2.2 Unpack ................................................................................................................... 20
3.3 Installation ......................................................................................................................... 22
3.3.1 Cleaning ................................................................................................................. 22
3.3.2 Check the Power Source ........................................................................................ 22
3.3.3 Plug In .................................................................................................................... 22
3.4 Testing ............................................................................................................................... 22
3.4.1 Signal to Noise Ratio Test ..............................
........................................................ 22
3.4.2
Wavelength Test ..................................................................................................... 23
4 Maintenance ................................................................................................................................. 24
4.1Routine maintenance .......................................................................................................... 24
4.2 Light Source Maintenance & Replacement ....................................................................... 24
7
4.2.1 Maintenance ........................................................................................................... 24
4.2.2 Light Source Replacement ..................................................................................... 24
Part IISoftware Manual ............................................................................................................... 25
5 Software Installation .................................................................................................................... 26
5.1 Requirements .................................................................................................................... 26
5.1.1 Hardware Requirements ......................................................................................... 26
5.1.2 System Requirements ............................................................................................. 26
5
.2 Install SPECTRO-97 Software ......................................................................................... 26
6 How To Use The Software ........................................................................................................... 31
6.1 Before Use ......................................................................................................................... 31
6.1.1 Connect to PC ........................................................................................................ 31
6.1.2 Link Procedure ....................................................................................................... 31
6.2 Functions ........................................................................................................................... 32
6.2.1 Measurement Modes .............................................................................................. 32
6.2.2 Interface ................................................................................................................. 33
6.3Software Operation ............................................................................................................ 38
6.3.1 Wavelength Scan .................................................................................................... 38
6.3.2 Time Scan ............................................................................................................... 47
6.3.3 Quantitative Analysis ............................................................................................. 54
6.3.4 3D-Scan .................................................................................................................. 65
6.3.5 Synchronous Scan .................................................................................................. 75
6.3.6 MORE CONVENIENT OPERATING METHODS .............................................. 83
PART IIIAPPENDIX .................................................................................................................. 95
Appendix I: Fluorescence & Phosphorescence ....................................................................... 96
F1.1 Theory ..................................................................................................................... 96
F1.2 Fluorescence analysis .............................................................................................. 99
Appendix II MEASUREMENT OF INSTRUMENTAL RESPONSE ............................ 100
F2.1 Theory ................................................................................................................... 100
F2.2 Measurement of Instrumental Response on Excitation Side ................................. 100
F2.3 Measurement of Instrumental Response on Emission Side .................................. 102
Appendix III Raman Scattering of Water & Detection Limit of Quinine Sulfate ............ 103
F3.1 Raman Scattering of Water ................................................................................... 103
F3.2 Water Raman Scattering S/N Ratio ...................................................................... 103
F3.3 Detection Limit of Quinine Sulfate ....................................................................... 104
Appendix IV Quantitative analysis wavelength method .................................................. 105
F4.1 Single Wavelength ................................................................................................ 105
F4.2 Double Wavelengths ............................................................................................. 105
F4.3 Triple Wavelengths................................................................................................ 106
Appendix V DETAILS ON QUANTITATIVE ............................................................... 107
F5.1 Linear Working Curve (1st order) ........................................................................ 107
F5.2 Quadratic Working Curve (2nd order) .................................................................. 107
F5.3 The correlation coefficient .................................................................................... 108
Appendix VI Synchronous Scan ...................................................................................... 109
F6.1 Constant Wavelength Difference .......................................................................... 109
F6.2 Constant Energy Difference .................................................................................. 109
8
Appendix VII Derivative Operation on Spectrum............................................................ 110
Appendix VIII Smoothing ................................................................................................ 112
F8.1 Savitzky–Golay ..................................................................................................... 112
F8.2 Mean ..................................................................................................................... 112
F8.3 Median .................................................................................................................. 1 1 2
Appendix IX Phosphorescence ........................................................................................ 113
F9.1 Theory ................................................................................................................... 113
F9.2 Phosphorescence Wavelength Scan ...................................................................... 113
F9.3 Phosphorescence Time Scan ................................................................................. 114
F9.4 Applications .......................................................................................................... 114
Appendix X Chemiluminescence ..................................................................................... 115
Appendix XI Multiple Excitation Scattering ................................................................... 116
Appendix XII Accessories ............................................................................................... 117
9
PART IUser’s
Guide
10
1 Appearance & Performance
1.1 Appearance
1.1.1 Body
Top Air-vent
Left Air-vent
Power Switch
Sample Compartment
Cover
Pilot Light
Panel
Fig.1-1 Body
Top Air-vent:Vent above the xenom light for air circulating. Please don’t touch. Left Air-ventVent beside the xenom light for air circulating. Power SwitchPlease turn to chap. 1.1.3 for more information. Sample Compartment CoverThe cover can be opened up to less than 90 degrees. Pilot Light:Power light.
11
1.1.2 Interface
Fig.1-2 Side
View
Fig.1-3 Interface
RS232 Serial
Port
USB Port
USB PortLink to PC with a USB cable. RS232 Serial PortFor debugging.
12
1.1.3 Power & Switches
Fig.1-4 Side
View
Fig.1-5 Power & Switches
Power
Plug-in
Xenom
Light
Main Power
Switch
Main Power
Fuse
Xenom Light
Switch
Power Plug-inFor connecting power cable. Xenom Light FuseFor xenom light fuse. Xenom Light SwitchTurn on/off the xenom light. Main Power Fuse:For main power fuse. Main Power Switch:Turn on/off the instrument.
13
1.1.4 Sample Compartment
1-4F97XP 荧光分光度计主机
Fig.1-6 Sample Compartment
Compartment
Cover
Sample
Hold
Fig.1-7 Quartz Fluorescence Sample Pool
Sample Compartment:Sample hold inside. Sample Hold:For fixing sample pool.
14
1.2 Mode of Operation
1.2.1 Signal Processing & Control System
Fig.1-8 Signal Processing & Control System
Controlled
Wavelengt
Xenon Light
Excitation
monochromator
Sample
Emission
monochromator
Photomultiplier
Pre-amplifier
Processor
USB
PC
Negative
High
Voltage
Xenon Light
Power
Excitation
Monitor
Opitcal
Gate
15
1.2.2 Light Path
Fig.1-9 Light Path
Lens
Lens
Lens
Len
Aspherical Mirror
Slit
Slit
Gratin
Sampl
Mirror2
Slit
Transflective
Mirror
Mirror3
Mirror4
Concav
e
Slit
Photomultiplier
Xenon
Light
Photocell
16
1.3 Functions
1.3.1 Modes For Measurement
1. Wavelength scan. Excitation wavelength scan provides spectrums of
fluorescence intensity which changes along with excitation wavelength under
fixed emission wavelength. Emission wavelength scan provides spectrums of
fluorescence intensity which changes along with emission wavelength under
fixed excitation wavelength.
2. Time scan. Time scan provides spectrums which changes along with time under
fixed emission wavelength and excitation wavelength.
3. Quantitative Analysis. According to the fluorescent spectra photometry, the
fluorescence intensity(F) is proportional to the concentration of the test sample(C)
under given conditions (the test group is dilute solution). Use fluorescent power F
and known sample concentration C to get standard curve. Then measure the
fluorescence of unknown sample to get the sample concentration.
4. 3D Scan. 3D scan includes excitation wavelength, emission wavelength and
fluorescence information.
5. Synchronous scan. The excitation side and the emission side scan at the same
time.
1.3.2 Self Tests & Adjustments
1. Self Tests & Adjustments
The instrument will initialize and self test while booting, including connection
detection, database detection, AD module detection, signal gain detection,
motion parts detection etc. Test result will be displayed on the screen and easy
for users to find problems.
1.4Performance
Tab.1-1 Performance of SPECTRO-97 series
Item Content
Excitation Source
150W xenon lamp(Hamamatsu)
Excitation
Wavelength
200nm~900nm
Emission Wavelength 200nm~900nm
Excitation Slit
SPECTRO-97XP/SPECTRO-97Pro2nm5nm10nm20nm
SPECTRO-9710nm
17
Emission Slit
SPECTRO-97XP/SPECTRO-97Pro2nm5nm10nm20nm
SPECTRO-9710nm
Wavelength Accuracy
SPECTRO-97XP±0.4nm
SPECTRO-97/Pro SPECTRO-97±1.0nm
Wavelength
Repeatability
SPECTRO-97XP≤0.2nm
SPECTRO-97/SPECTRO-97Pro≤0.5nm
Signal-to-Noise Ratio
SPECTRO-97XP:Raman peak of water(P-P):S/N≥200(10nm Slit
SPECTRO-97/97Pro:Raman peak of water(P-P):S/N≥150(
10nm Slit
Limit
SPECTRO-97XP≤5×10
-11
g/ml(Quinine Sulfate Solution)
SPECTRO-97/97Pro≤1×10
-10
g/mlQuinine Sulfate Solution
Linearity γ≥0.995
Peak Repeatability ≤1.5%
Stability(10min)
Zero Drift±0.3
Value Limit±1.5%
Wavelength Scan
Speed
Multi-speed Level, Maximum at 48000nm/min
Photometric
Quantity Range
0.00-10000.00
Data Transportation USB2.0
Power 200W
Power Source
AC 220V/50Hz 110V/60Hz
Demension
380×445×310mm
Weight
Net Weight12kg Gross Weight14kg
18
2 Booting & Shutting Down
2.1 Booting Status
2.1.1 Booting
Put the instrument on steady platform. Make sure the main power and light power
switch are off(switch on 0). Plug in the power line.
There is a certain probability that the high voltage trigger of the Xenon light power
will affect other electrical equipment around. We highly recommend you check the
following status before turning on the Xenon light power:
1 Unplug the USB cable connecting PC and the instrument. 2 Make sure the grounding of power source is reliable. 3 If other instruments in the same platform were affected by the high trigger
voltage before, please shut down those instruments and turn on after the xenon
light is triggered.
4 Turn on the xenon light power, then turn on main power after the xenon light is
triggered.
Connect PC and the instrument with the USB cable, then run the PC software.
ATTENTION
If the xenon light is not triggered properly and making high noise,
shut down the xenon light power immediately. Please turn on the xenon
light after a few seconds.(It happens only when the power is not stable
or the xenon light is reaching its limit)
As xenon lamp life and the switching times are closely related,
Please reduce unnecessary trigger.
Xenon light needs 30 minutes to stable.
Don’t turn on Xenon light without turning on the main power after.
2.1.2 Fans Condition
Please make sure the fans on the left side and on the top left are functioning every
time startup. If the fans are not working, shut it down and check.
19
2.1.3 Multi-instruments Booting
When using multiple instruments, please turn on all the xenon lights first, then turn
on all the main power to reduce the influence of the high trigger voltage.
2.1.4 Initialization
Initialization status will be displayed on computer software as Fig.1.3.2-1.
2.2 Power Off
When connecting to PC, close the software first; then turn off the main power; at
last turn off the xenon light power.
ATTENTION: To restart the Xenon light power, please wait for 60
seconds after power off.
20
3 Installation
3.1Environment
The instrument is suitable for analysis in laboratory environment. For its work with
computers, so need to meet the following working condition.
3.1.1 Laboratory Environment
Temperature 1030 , humidity under 85%. Avoid corrosive gas and the organic
and inorganic gases which are absorptive within the range of ultraviolet.
3.1.2 Work Bench
The work bench should be smooth and solid. Avoid vibration, dust, direct sunlight.
3.1.3 Power
AC 220V±22V50Hz±1Hz or 110V±11V60Hz±1Hz.
3.1.4Environment Change
If the instrument is needed in the field, please make sure the environment meets the
requirements above. Please use the original package moving instrument. If there are
special requirements please inform us when ordering.
3.2 Package
The instrument adopts carton packaging. Long-distance transport may require
additional outside wooden box.
3.2.1 Check the Package
Before unpacking, make sure the packaging is intact. If the package is damaged,
please contact with the transportation insurance.
3.2.2 Unpack
Open the case and carefully take out the instrument (Please keep the package for
21
transportation). Make sure the instrument and all accessories are correct according to
the package list. Please contact us if there is any mistake.
Table.3-1 Fluorescence spectrophotometer package list
Standard
fittings
SPECTRO-97 series instrument One piece
Fluorescence spectrophotometer
software
One set
Power Cable One piece
USB Cable One piece
Quartz fluorescence cell 10mm One piece
Fuse(2A/5A) two pieces
Instruction Manual for SPECTRO-97
series
Fluorospectrophotometer
One copy
Applications References Manual
One copy
Certification of products One copy
Packing list One copy
Guarantee repair list One copy
Dust proof One piece
Optional
spare
parts
Quartz fluore
scence cell 10mm
Glass fluorescence cell 10mm
Fuse(2A/5A)
USB cable
Power cable
Optional
accesso
ries
Cut-of
f filter
PC
Membrane kind sample accessories
Powder kind sample accessories
Micro scale capillary sample accessories
Jacket sample pool accessories
200μL centrifuge tube accessories
Note: Optional spare parts and accessories will be delivered according to your
shipping contract.
22
3.3 Installation
3.3.1 Cleaning
Remove the tape and clean the surface.
3.3.2 Check the Power Source
Make sure the instrument power supply voltage and area voltage are correct.
3.3.3 Plug In
Put the instrument on a stable work table about 10 cm away from wall. Plug in the
power cable to the lab power.
3.4 Testing
3.4.1 Signal to Noise Ratio Test
1. Turn on the instrument and preheat. Light source and electronic components require to reach heat balance after startup.
Start operation after 30 minutes preheating.
2. Put in sample Choose a clean quartz fluorescence cell. Fill the quartz fluorescence cell with sample
(double distilled water). Then put it in the sample cell.
ATTENTION: Dirty quartz fluorescence cell will affect the accuracy
of the test.
3. Run emission wavelength scan
Choose “Wavelength Scan” –“Emission wavelength scan”. Set the excitation wavelength at 350nm, the emission wavelength at 300nm to 500nm. Set slit to 10nm. Set scan speed at 60nm/min. Response: auto. Gain: high. Run wavelength scan, then check the peak around emission wavelength 397nm. This peak is signal S.
4. Run time scan.
Choose “Time scan”. Set the excitation wavelength at 350nm. Set the emission wavelength at 397nm. Scan time 120 seconds. Slit 10nm. Response time 2 seconds. When the signal is stable(Let doubly distilled water expose more than three minutes above conditions), run time scan.
5. Calculate
Note the peak signal S and the Peak-valley value N. S/N is the Raman signal to noise ratio.
23
Attention
The energy of Raman is weak, so the measurement is easy to be
interfered. If the result looks bad, please test again.
In order to protect the PMT tube, when the gain is high(above 6), please
don’t put high energy light source into the sample cell.
Setting the gain will affect the fluorescent zero. Please reset zero after
changing gain.
3.4.2 Wavelength Test
The instrument will automatically adjust wavelength. If there is any wavelength mistake,
trained professionals are allowed to test wavelength with mercury-arc lamp or
fluorescent lamp. Learn the test method through training.
Mercury-arc lamp spectral line wavelength and figure below.
Table.3-2Emission spectra of low pressure mercury lamp in UV and visible region
NO. Wavelength/nm NO. Wavelength/nm
1 253.65 7 404.66
2 296.73 8 407.78
3 302.15 9 435.84
4 313.16 10 546.07
5 334.15 11 576.96
6 365.01 12 579.07
Fig.3-1Spectra of mercury-arc lamp
Power
24
4 Maintenance
4.1Routine maintenance
1. Always check whether it meets the requirements of the work environment in daily use.
2. Keep the vents functioning while the instrument is on.
ATTENTION: High temperature vents. Keep distance.
3. Keep the instrument clean. Add a dust cover when not in use. Use water to clean the instrument appearance. DO NOT use alcohol, ether, acetone and other organic solvents. Do not clean when the instrument is working.
4. Keep the quartz fluorescence cell clean.
4.2 Light Source Maintenance & Replacement
4.2.1 Maintenance
1. Keep the light source clean.
2. Strictly in accordance with the order of operations when turn on/off xenon light.
ATTENTION: When power on, turn on the xenon light power first,
then the main power. When power off, turn off the main power first, then
the xenon light power.
3. Xenon light will be hard to trigger when the power is not stable or the xenon
light is reaching its limit.
ATTENTION: If the xenon light is not triggered properly and making
high noise, shut down the xenon light power immediately. Please turn on
the xenon light after a few seconds.
4. Avoid repeated triggering xenon lamp.
As xenon lamp life and the switching times are closely related,
Please reduce unnecessary trigger.
5. Make sure that the instrument cooling fan is working properly. Make sure the
surface of the instrument top vents maintain good ventilation.
4.2.2 Light Source Replacement
Professionals are allowed to replace the light source. Specific methods and
calibration procedures will be introduced through training.
25
Part IISoftware
Manual
26
5 Software Installation
Before reading this section, please read Part I carefully. Windows
XP system is recommended.
In order to run the software properly in Windows 7, please run the
......................................................
software in administrator account.
...............................
5.1 Requirements
5.1.1 Hardware Requirements
Hardware Minimum requirements
CPU Intel P4 2.0GHz or same level CPU
Memories 512M
Hard disk No less than 200M disk space
USB USB2.0
CD drive CD-ROM
Monitor resolution 1024*768
16-bit color
Table.5-1 Hardware requirements
5.1.2 System Requirements
Windows XP or higher version is recommended. Please turn off the screen saver
and
power management program while SPECTRO-97 Fluorescence Spectrophotometer
data processing software is running.
5.2 Install SPECTRO-97 Software
Put the software CD into the drive, the application will automatically start.
27
If the installation does not start, open X:\SETUP.EXE(X is the CD drive)
as Fig.5-1.This manual is based on SPECTRO-97 Pro.
Fig.5-1 Installation
Click “Next” in Fig.5-1, input user name and company name.
Fig.5-2 Information
Click “Next” in Fig.5-2 to set install path. Default path is as in Fig.5-3: C:\Program
28
File\MRC\SPECTRO-97Pro Fluorescence spectrophotometer. Click “Change”
to change the path. Click “Back” to go back, click “Cancel” to cancel.
Fig.5-3 Install path
Click “Next” to set account limit and shortcut path in fig.5-4.
Fig.5-4 Shortcut path & account limit
29
Fig.5-5 Installation information
Click “Next” if all the information is correct to start installation.
Fig.5-6 Installing
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