BioLAB BSFL-103 User manual

Operation Manual
Fluorescence Spectrophotometer
BSFL-103
Thank you for Choosing Biolab products. Please read the “Operating Instructions” and
“Warranty” before operating this unit to assure proper operation.

Index
. Preface.................................................................................................................................................03
2. Appearance and Performances of the Instrument .............................................08
3. Operation of the Instrument....................................................................................................... 4
4. Installation of the Instrument...................................................................................................... 9
5. Maintenance of the Instrument..................................................................................................24
6. Environmental Requirements of Using Software and Installation.................................26
7. Operation of Software.....................................................................................................................27
8. Operation of Wavelength Scan Mode (Connected with a Computer).........................3
9. Operation of Time Scanning Mode (When Connected with a Computer).................39
0. Operation of Quantitative Analysis Mode (When connected with a Computer)......42
. Appendix: Data Processing with Microsoft Excel 2000.......................................................50
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Fluorescence Spectrophotometer BSFL-103

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 BSFL- 03 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 BSFL- 03 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 2 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
!
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Fluorescence Spectrophotometer BSFL-103
Symbol indicates important information of operating the instrument
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.

Points for Attention to Operation
. The BSFL- 03 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.
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
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pass into the sample compartment. When a sample of unknown concentration is measured,
PMT level should be set from low grade to high grade ( - 7) 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
“BSFL- 03pc.exe” process, then restart the software and the instrument.
0. Within the monochromator the tightened screws must not be loosened. The optical
devices and the working environment of the instrument should be kept clean.
. 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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Fluorescence Spectrophotometer BSFL-103

0 Appearance and Performances of the
Instrument
. Appearance of the Complete Instrument and Main Components
. 38% The Complete InstrumentThe Complete Instrument
Fig. - BSFL- 03 Fluorospectrophotometer Main Unit
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Heat Dissipation Opening at the Top of the Instrument
Heat Dissipation Fan
Lid of Sample
Compartment
Power Line
Operating Panel
USB Interfa e
RS232 Communi ation Conne tor

Fig. -2 The Left Side View of BSFL- 03 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).
Main Components
1. Xenon Lamp
Fig. -3 Xenon Lamp and Lamp Holder
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Fluorescence Spectrophotometer BSFL-103
Line
Xenon Lamp (The
lightening part)
Lens
Line

2. Sample Compartment
Fig. -4 Sample Compartment and Quartz Sample Cell ( 0 mm)
3. Filter
Fig. -5 Filter (center wavelength 365 nm)
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Filter Position
Sample Cell
Position

1
2 3
4
5
1.2 Operating Panel of the Instrument
Fig. -6 Operating Panel of BSFL- 03 Fluorospectrophotometer Main Unit
. key, 2. key, 3. key, 4. key, 5. Display window.
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Fluorescence Spectrophotometer BSFL-103
Mode Print Print Normalise

1.3 Working Prin iples of the Instrument
Signal Pro essing and Control System
Fig. -7 Signal Processing and Control System
Opti al System
Fig. -8 Optical System Arrangement of BSFL- 03 Fluorospectrophotometer
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Xenon
Lamp
Power Sour e of
Xenon Lamp
Ex itation
Filter
Sample Cell USB
Communi ation Conne tor
Photomultiplier Emission
Mono hromatoraton
单色器
Monolithi
Pro essor
Display
Key
Board
Serial Interfa e
Serial Printer
Preamplifier
Negative
High Voltage
Computer
Controlled
Wavelength

1.4 Measurement Fun tions of the Instrument
1.4.1 Working Modes of Measurement
1. Wavelength s an mode. At a fixed excitation wavelength a spectrum is obtained in which
the fluorescent intensity of a sample varies with wavelength.
2. Time s an 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
. 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 omputer. 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.
4.3 Self- he king and Self- orre ting
. 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 -2. Wavelength
correcting program will guarantee the accuracy of wavelength.
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Fluorescence Spectrophotometer BSFL-103

1.5 Performan e Index of the Instrument (Spe ifi ations)
Light source 50W 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 ± nm
Wavelength repetition ≤0.5nm
Emission spectrum bandwidth: 0nm
S∕N ratio S/N≥ 50 (Using cm quartz sample cell, measure the signal noise
ratio
of Raman spectrum of water )
Check utmost × 0- 0 g/ml sulfuric acid quinine solution
Measure linearity ≥0.995
Peak value ≤ .5%
Zero line drift ≤0.3% (Within 0 minutes)
Upper limit change
of indicating value
± .5% (Within 0 minutes)
Power requirement 220V±22V 0V/60HZ
Dimensions 442×392×250（mm）
Weight Net weight kg
Gross weight 3kg
Table - Performance Index of the Instrument
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02 Operation of the Instrument
2.1 Operation of the Instrument Panel
1 Display Window
The display window is a 28*64 pixels LCD, shows information including initialization status,
fluorescence value, concentration value, emission wavelength value, PMT level, concentration
factors, standard concentration, etc.
Symbol related:
. “●” means the excitation light source is on.
2. “%” means the fluorescence value has been normalized.
2 Operation of the Fun tion Keys
. “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 Level Adjust, 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 00
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.
2.2 Normally Working Condition of Startup
2.2.1 Startup of the Instrument
First turn on the power source of the xenon lamp, and then turn on the power source of the
BSFL- 03 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 :
If the xenon lamp fails to be lit and a squeaking sound of high frequency or a cracking ①
sound of sparking is continuously produced, please turn off the power source of xenon lamp
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Fluorescence Spectrophotometer BSFL-103

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).
Because the life of the xenon lamp is closely related to the number of on-off of the lamp ②
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 -2.
STEP PASS FAILURE
Self-
checking
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
Motor Module Motor Module initialized Motor error
Motor driver board error
Self-
correcting
Wavelength
correcting
The 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 -2 BSFL- 03 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 BSFL- 03 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 pro edures 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. .2 for
detail). The wavelength will be automatically set to target value.
(2) PMT Level Adjustment. 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
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Fluorescence Spectrophotometer BSFL-103

Fluorescence Value Mode. Wait till the value is stable. Press “Adj 0” to adjust the fluorescence
value to zero(±0. ).
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” provided by Appendix.
(5) Measurement for Unknown Sample. Place the solution of the unknown sample in the
light path. After the fluorescence value is stable, record the value. Acquire the concentration
of the unknown sample from the working curve equation.
2. Measuring Pro edures by a Single-point Method of Standard Sample
Prepare one standard solution and one blank solution each. After finishing the preparation of
unknown sample solution, operate as the following sequence.
Note: The concentration of standard sample should close to that of sample to be measured as
far as possible.
(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. .2 for
detail). The wavelength will be automatically set to target value.
(2) PMT Level Adjustment. 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 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
value to zero(±0. ).
Attention: Zero adjustment is not allowed in the following measurement.
(4) Dire t-reading of Con entration. Press the “Mode” key to enter Concentration Input
Mode. When the LCD shows “Input Concentration: ”, use “Adj 0” key or the “Normalize” key to
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adjust the value to be the concentration of standard solution. Then press the “Print” key to
calculate the concentration factor and switch to Concentration Mode.
(5) Measurement for Unknown Sample. Place the unknown sample in the sample cell. The
value on the display window is the concentration of the unknown sample in Concentration
Mode. Press the “Print” key to print the current data if connected to a dedicated serial
interface printer.
(6) Readout of Con entration Fa tor. Press the “Mode” key to switch Factor Input Mode.
When the LCD shows “Input Factor: ”, the value is the current concentration factor. Use “Adj 0”
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
Turn off the power source of the BSFL- 03 Fluorospectrophotometer main unit first, and then
the power source of the xenon lamp. (The order is reverse to that of startup.)
If in on-line mode, close the software first, then turn off the power source of the main unit,
and turn off the Xenon lamp.
Note: After turning off the power source of the xenon lamp, please wait for 60 seconds at
least. Then turn on the xenon lamp if you want to use it once again.
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Fluorescence Spectrophotometer BSFL-103

03 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 Spa e Environment of Laboratory
Environment temperature range: 0-30 . Environment humidity is less than 85℅.Corrosive gas℃
and other organic or inorganic solvent gas that is absorptive within the range of violet
radiation should not be contained in the atmosphere.
3.1.2 Working Table
The instrument should be placed on a sturdy and level table or the like, avoiding strong
vibration, direct sun light, and dust.
3.1.3 Power Sour e
Alternating voltage must be within a range of 220V±22V, 0V± V.
3.1.4 Environmental Alternation
If the instrument needs to be used at a worksite, the working environment should accord with
the above demands for environment. If moving the instrument to other places, please use
original packing materials. If users have any special demands, may place a special order for
the instrument.
3.2 Unpa king for Che king
The BSFL- 03 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.
3.2.1 Che king the Outer Pa king
Before unpacking, check that the outer packing is intact. If finding the packing to be
incomplete or finding marks of impact on the packing, please contact the transport insurance
department.
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3.2.2 Unpa king for Counting
Unpack at the position of adhesive seal tape. Take out the BSFL- 03 Fluorospectrophotometer
main unit carefully. (Please keep the outer packing so as to use it again when moving the
instrument later. Check the main unit, the standard fittings, accessories purchased by choice
and spare parts against the packing list. If anything is missing or damaged, please contact the
local dealer or our company immediately.
Standard
fittings
BSFL- 03 Fluorospectrophotometer main
unit
One
piece
365nm filter (installed in the main unit) One
piece
Software package of data processing for
F96 Pro Fluorospectrophotometer (set) One set
Power line One
piece
USB wire One
piece
Quartz sample cell 0mm One pair
Fuse (2A/5A) two
pieces
Instruction Manual for BSFL- 03
Fluorospectrophotometer One
copy
Certification of products One
copy
Packing list One
copy
Spare
parts
Purchase
d by
choice
200―850nm Interference filter (φ25nm)
Quartz sample cell 0mm
Glass sample cell 0mm
Fuse (2A/5A)
USB wire
Power line
Accessories
Purchased
by choice
Microcomputer main unit
Display device
Dedicated serial interface printer
Membrane kind sample accessories
Powder kind sample accessories
Microscale capillary sample accessories
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