MRC Spectro-97 Series User manual

Fluorescent Spectrophotometers
SPECTRO-97 SERIES
Operation Manual
PLEASE READ THIS MANUAL CAREFULLY BEFORE OPERATION
Hagavish st. Israel 58817 Tel: [phone], Fax: [phone] [email protected]
MRC.4.18

1
Preface
Thank you for purchasing the Fluorospectro-
photometer of MRC Ltd. Please read this manual
carefully before installation or first time
using SPECTRO-97 series
Fluorospectrophotometer.
Fluorometry is 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 fluorospectrophtometer
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
Please 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 on 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, turn
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 the
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 top
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 other

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 I：U 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 II：Software 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 III：APPENDIX .................................................................................................................. 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 I：User’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-vent：Vent beside the xenom light for air circulating.
Power Switch：Please turn to chap. 1.1.3 for more information.
Sample Compartment Cover：The 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 Port：Link to PC with a USB cable.
RS232 Serial Port：For 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-in：For connecting power cable.
Xenom Light Fuse：For xenom light fuse.
Xenom Light Switch：Turn 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
图1-4F97XP 荧光分光光度计主机侧图
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
Mirror1
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-97Pro：2nm、5nm、10nm、20nm
SPECTRO-97：10nm

17
Emission Slit
SPECTRO-97XP/SPECTRO-97Pro：2nm、5nm、10nm、20nm
SPECTRO-97：10nm
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/ml（Quinine 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 380×445×310（mm）
Weight Net Weight：12kg Gross Weight：14kg

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：
①If 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)
②As xenon lamp life and the switching times are closely related,
Please reduce unnecessary trigger.
③Xenon light needs 30 minutes to stable.
④Don’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.
！