Nano-100
Micro-Spectrohpotometer
USERS MANUAL
Version 1.0
HANGZHOU ALLSHENG INSTRUMENTS CO., LTD.
Foreword
Thank you for purchasing our Products: Micro-Spectrohpotometer. This
Manual for users contains function and operation of the Instrument. In order
to use the instrument properly, please read this manual carefully before
using the Instrument. Keep it for later use when you meet with difficulties.
Opening Check
Please check the Instrument and Appendix with the packing list when you
first open the instrument packing case. If you find there is something wrong
with the Instrument and the Appendix, do contact the vendor or the
producer.
HANGZHOU ALLSHENG INSTRUMENTS CO., LTD.
ADD：
No.2, Xiyuan Road 6th, Economic Park of Science and Technology
of the Westlake, Town of Sandun, Hangzhou, China
Tel.：
+86-571-88802738
Fax:
+86-571- 87205673
E-mail: [email protected]
Website: www.allsheng.com
File No.：AS64SM
Version No.：Version 1st, Nov. 2011
Safety Warnings and Guidelines
1 Important operation information of the security:
Before the users’ operation, they should have a perfect conception of how to use
the Instrument. Therefore, read this Manual carefully before using it.
Operation before reading the Manual is forbidden. Read the guidelines
and directions below and carry out the countermeasure according to
them.
2 Security:
The operation, maintenance and repair of the Instrument should comply with the
basic guidelines and the remarked warning below. If you don’t comply with them, it
will have effect on the scheduled using life of the Instrument and the protection
provided.
This product is indoor Instrument.
Read the Manual carefully before operation, or it may cause injured.
The expert of wiring equipment can operate this Instrument.
The operator should not open or repair the Instrument by himself, which
will result in losing the qualification of repair guarantee or occur
accident. If there is some wrong with the Instrument, the company will
repair it.
Power off when you finish your work. Pull off the connector plug when
there’s long time no use of the Instrument and cover it with a cloth or
plastic paper to prevent from dust.
Pull the connector plug from the jack at once in the following case, and
contact the vendor:

There is some liquid flowing into the Instrument;

Drenched or fire burned.

Abnormal operation: such as abnormal sound or smell.

Instrument dropping or outer shell damaged.

The function has obviously changed.

3
The maintenance of Instrument
The pedestal should be cleaned by the cloth stained with alcohol.
If there are smutches on the Instrument, clean them with cloth stained with
alcohol.
CONTENTS
Chapter 1 Introduction------------------------------------------------------------------1
Chapter 2 Specifications---------------------------------------------------------------2
1 The normal operating condition-----------------------------------------------2
2 The basic parameters and performance------------------------------------2
Chapter 3 Test Principle----------------------------------------------------------------3
Chapter 4 Initial Setup -----------------------------------------------------------------6
Chapter 5 Software Operation-------------------------------------------------------11
Nano-100 Operation Manual
Chapter 1 Introduction
Chapter 1 Introduction
The Nano-100 is a full-spectrum (200-850nm) spectrophotometer that measures
0.5-2ul samples with high accuracy and reproducibility. It utilizes a patented sample
retention technology that employs surface tension alone to hold the sample in
place. This eliminates the need for cumbersome cuvettes and other sample
containment devices and allows for clean up in seconds. In addition，the Nano-100
has the capability to measure highly concentrated samples without dilution(100X
higher concentration than the samples measured by a standard cuvette
spectrophotometer).
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Chapter 2 Specification
Chapter 2 Specifications
1. The normal operating condition
Ambient temperature：5C  35C
The relative humidity：≤70%
Power Supply：DC24V 2A
2. The basic parameters and performance
Model
Parameter
Nano-100
Minimum Sample Size
0.5ul-2ul (2ul advised)
0.2mm or 1mm
Path Length
Light Source
Xenon flash lamp
Detector Type
3864—element linear silicon CCD array
Wavelength Range
200－850nm
±1 nm
Wavelength Accuracy
≤3nm（FWHM@Hg 253.7nm）
Spectral Resolution
Absorbance Precision
0.003Abs（1mm path length）
Absorbance Accuracy
1％（at 0.76 at 350nm）
Absorbance Range
0.02 - 75（10mm equivalent）
Detection limit
8ng/ul dsDNA
Maximum Detection Concentration
5,000ng/ul（dsDNA）
Detection Time
10 - 20s
Dimension (mm)
200×262×154mm
Net Weight (kg)
2.5 kg
Pedestal Material
SUS304
Operating Voltage
24V
Power
12-18W
Software Compatibility
Windows XP、Vista（32bit）and Win7（32bit）
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Nano-100 manual
Chapter 3 Test Principle
Chapter 3
Test Principle
1. Samples Retention System
A 2ul sample is pipetted onto the end of a fiber optic cable. It is at least to use 0.5ul
sample to measure high concentration nucleic acid and protein A280. A second fiber
optic cable (the source fiber) is then brought into contact with the liquid sample causing
the liquid to bridge the gap between the fiber optic ends. A pulsed xenon flash lamp
provides the light source and a spectrometer utilizing a linear CCD array is used to
analyze the light after passing through the sample. The instrument is controlled by
special software run from a PC, and the data is logged in an Excel file on the PC.
2. Sample Size Requirements
Although sample size is not critical, it is essential that the liquid column be formed so
that the gap between the upper and lower measurement pedestals is bridged with
samples
The hydrophobic between the water molecules is the main factor of surface tension. In
general, the presence solute of liquids (（including protein, DNA,RNA, salt ion, detergent
molecule) can significantly reduce surface tension. Although, for most samples, a 1ul
sample size is enough, a 2ul sample size is recommended for protein measurements.
Field experience indicates that the following volumes are sufficient to ensure
reproducibility:
Aqueous solution of nucleic acid：1ul
Purified protein：2ul
Microbial cell suspension：2ul
Others：2ul
It is best to use a precision pipettor (0-2ul) with precision tips to assure the
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Chapter 3 Test Principle
sufficient sample ( 1-2ul) is used. Lower precision pipettors (0-10ul and larger) are
not as good as at delivering 1 ul volumes to the measurement pedestal. If you are
unsure about your sample characteristics or users or pipettor accuracy, a 2 ul sample is
recommended.
3. Basic use
3.1 With the sampling arm open, pipette the sample onto the lower measurement
pedestal.
3.2 Close the sampling arm and initiate a spectral measurement using the operating
software on the PC. The sample column is automatically drawn between the upper and
lower measurement pedestals and the spectral measurement made.
3.3 When the measurement is complete, open the sampling arm and wipe the sample from
both the upper and lower pedestals using a soft laboratory wipe. Simple wiping
prevents sample carryover in successive measurements for sample.
4. Blanking and Absorbance Calculation
When NANO-100 Spectrophotometer is “blanked”, a spectrum is taken a reference
material (blank) and stored in memory as an array of light intensities by wavelength.
When a measurement of a sample is taken, the intensity of light that has transmitted
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Chapter 3 Test Principle
through the sample is recorded. The sample intensities along with the bland intensities
are used to calculate the sample absorbance according to the following equation.
Thus, the measured light intensity of both the sample and of the blank are required to
calculate the absorbance at a given wavelength. The Beer-Lambert equation is used to
correlate the calculated absorbance with concentration.
A＝absorbance represented in absorbance units (A)
ε＝the wavelength—dependent molar absorptivity (unit: L/mol*cm)
b＝path length (unit: cm)
c＝sample concentration (unit: mol/L)
Contrasted solution, or blank solution ，is normally solvent to dissolve the targeting
molecular and it need to be same with the samples in PH and ionic strength
5. Blank Cycle
For most consistent results, it is best to begin any measurement session with a blanking
cycle. This will assure the user that the instrument is working well and that the pedestal
is clean. To perform a blanking cycle, perform the following:
1. Load a blank sample onto the lower measurement pedestal and lower the sampling
arm into the 'down' position.
2. Click 'Blank' button to make a blank and save the reference spectra.
3. Analyze an aliquot of the blanking solution as though it were a sample. This is done
using the 'Measure' button. The result should be a spectrum with a relatively flat
baseline, and the absorbance value change should be within 0.04A(10mm path).
4. Wipe the both pedestals and repeat the process until the spectrum is within 0.05A
(10mm path).
Although, there is no need to make a blanking calculation between every sample, it is
best to make a blanking calculation every 30min when measure several samples. After
30min, the time for making last blanking will display in the status bar of the software.
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Nano-100 Operation Manual
Chapter 4 Initial Setup
Chapter 4 Initial Setup
1. Installation requirement：
Computer requirements：
Microsoft Windows XP、Win7（32bit）or Vista（32bit）operating system.
1.5GHz or higher processor.
1GB or more of RAM（2GB for Vista operation system）.
100MB of free hard disk space.
Open USB port（the instrument can only be connected via the USB port）.
Microsoft office 2003.
2. Installation preparation：
The system software must be loaded onto the PC before the USB cable is connected.
Administrator access on the PC is required to install the software.
3. Installation steps:
3.1 Close all programs and make sure that USB cable is unplugged.
3.2 Insert the operation software CD in the CD drive of the PC. Click
“
” and then enter into the installation step
Step1：Select Setup Language:
Step2：Click “Next” twice and then input the password supplied by the manufacturer
(initial password:123456)
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Chapter 4 Initial Setup
Click “next”。
Step3：Select Destination Location
Step4： Select shortcut.
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Chapter 4 Initial Setup
Step5：select default setting until the installation ends.
Click the “Finish” button and begin the installation.
3.3 Connect the USB cable to instrument and the computer (installed with software). Turn
on the power switch; the red indicator will be bright.
Normally, the computer will remind the user to find a new hardware Dialog box. In this
time, choose the last item, and click “Next” to enter into next step.
Then select “Install from a list or specific location (Advanced)”.
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Chapter 4 Initial Setup
3.4 Finally, Input “Nano 100 Drive” folder’ path, such as “C:\Program Files\Nano 100\ Nano 100
Drive” in “Include this location in the search” and then click “Next”. The drive will start
installation.
3.5 After installation, there will be a prompt Dialog box
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Chapter 4 Initial Setup
If the computer is prompted to install the driver again, repeat the above process. The computer
equipment manager will have the following tips after driver installation is successful.
If the Device Manager still display “?”, it means that this device hasn’t been installed properly.
Select the device again and click the right mouse button to reinstall the device.
Then the user can open the running test software.
At the same time, the user can see “.Net 2.0” frame in the “Add or Remove Programs”.
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Nano-100 Operation Manual
Chapter 5 Software Operation
Chapter 5 Software Operation
1. General introduction
1.1 USB cable connection
To make measurements with the instrument, connect the USB cable to instrument and the
PC, plug in the 24V power supply and connect to the power input at the back of the
instrument. The power supply can remain plugged into the Nano-100 while the instrument
is not in use. The unit is in “standby” mode, power consumption is 5W and the flashlamp is
not energized. After plugging tin the 24V power supply, utilize a power switch, there will be
a red indication in the top of the machine. The green light indicates that the unit is working.
1.2 Software features
There are two parts in Nano-100 software, status bar and working key in the left part and
display data window in the right part.
（1） Taskbar
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Chapter 5 Software Operation
The taskbar options include the following options:
Home – display the application main
My Date- restore the sample data saved in the folder specified by the user
Diagnostics- test instrument if it is connected properly
Options- enter into Debug mode
(2) Application selection region
Click to select the (color-keyed) type of the sample, and then enter the selected type interface.
(3) Function key
When the “application” is opened, the following five function keys are displayed in the top of
left column:
Blank - Use dissolved sample buffer to make a blank. Before making a sample measurement,
a blank must be measured.
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Chapter 5 Software Operation
Measure – Initiate the measurement for the samples
To Excel – The detail absorbance data of present measurement is stored in the Excel
table which is specified by user.
To Picture –To store the software (the user is working) interface image
Print – Print a copy of current data and corresponding spectrum to the default printer.
（4）Main menu bar option
File – The dropdown menu of file menu includes the following options:
New - The menu for application operated by specific user group, equivalent to the “Home”
key.
Open- Restore the sample data saved in the folder specified by the user, equivalent to “My
Date” key.
Save - The detail absorbance data of present measurement is stored in the Excel table
which is specified by user. It is equivalent to “To Excel” Key.
Exit – Exit program
Tools – The dropdown menu of tools menu includes the following options:
Check correction –Test instrument if it is connected properly, it is equivalent to
“Diagnostics” key.
Dye Edit – Edit dye information.
Language - Select operation language.
Debug - The dropdown menu of debug menu includes the following options:
DebugEnable -Enter or exit the Debuge mode
Help – The dropdown menu of help menu includes the following options:
Contents - Display electronic version of the manual.
About- Display information about the software.
2. Applications
UV/VIS spectrophotometry is simple for small samples by using Nano-100 Spectrophotometer.
The small sample requirement and ease of use make the Nano-100 Spectrophotometer
ideally suited for measuring:
 Nucleic acid concentration and purity of nucleic acid samples up to 5000ng / UL (dsDNA)
without dilution
 General UV – Vis spectrophotometry
 Purified protein analysis(A280)
 Fluorescent dye density of Micro array samples
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Chapter 5 Software Operation
2.1 Quick start
(1) Double-click the software icon and select the interested application from the right
column
(2) Use suitable buffer to create a blank. Load a blank sample onto the lower
measurement pedestal and lower the sampling arm into the “down” position. Then,
click on the “Blank” button.
(3) When the measurement is complete, wipe the blanking buffer from both pedestals
using a laboratory wipe. Input the sample name in the suitable position and pipette 2ul
sample to make measurement.
Note: each sample must be pipette freshly
After measurement:
Wipe the sample from both upper and lower pedestals upon completion of each sample
measurement by using clean dust-free paper, so that it can do next sample measurement.
The concentration limit values of measurement as below:
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Chapter 5 Software Operation
3. Nucleic Acids measurement
3.1 General Information
Nucleic acid samples can be readily checked for concentration and quality using
Nano-100 Spectrophotometer. To measure nucleic acid samples select the ‘Nucleic Acid”
application module.
The Beer-Lambert equation is used to calculate nucleic acid concentration. Please see
following equation.
C＝nucleic acid concentration (unit: ng/ml)
A＝the absorbance in AU
ε＝the wavelength-dependent extinction coefficient (unit: ng-cm/ml)
b＝path length (unit: cm)
The generally accepted extinction coefficients for nucleic acids are:
Double-stranded DNA：50ng-cm/ul
Single-stranded DNA：33ng-cm/ul
RNA：40ng-cm/ul
When choosing pedestal mode, Nano-100 Spectrophotometer can measure the high
concentration without dilution by using 1.00mm and 0.2mm short path length to measure.
Note: The absorbance value of nucleic acid measurement is consistency of the reading
value under 1cm path length.
The Nano-100 spectrophotometer can accurately measure double-stranded DNA samples
up to 5000ng/ul without dilution. For each sample, software will automatically optimize the
best path length to make measurement. When the optical intensity (after measurement
sample extinction) is lower than 200(under 1cm path length), software will inform the
customer to choose shorter path length to make sure the precision of the measurement.
Unique screen is shown as below.
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Chapter 5 Software Operation
The data shown in the spectrum image are consistency of the reading value under 10mm
path length.
The right column of spectrum includes following information:
Name－Input the sample name: when sample measurement, the sample name should be
input.
Type－Used to select the type of nucleic acid being measured. The user can select
‘DNA-50’ for dsDNA, ‘RNA-40’ for RNA, ‘ssDNA-33’ for single-stranded DNA.
ID--Display the serial number for the sample being measured. The user can open the
software to start recording.
A260－Absorbance of the sample at 260nm with 10mm path
A280－Absorbance of the sample at 280nm with 10mm path
260/280－Ratio of sample absorbance at 260nm and 280nm. The ratio of absorbance at
260 and 280nm is used to assess the purity of DNA and RNA. A ratio of ~1.8 is generally
accepted as “pure” for DNA; a ratio of ~2.0 is generally accepted as “pure” for RNA. If the
ratio is appreciably lower in either case, it may indicate the presence of protein, phenol or
other contaminants that absorb strongly at or near 280nm.
260/230－ratio of sample absorbance at 260 and 230nm. This is a secondary measure of
nucleic acid purity. The 260/230 values of “pure” nucleic acid are often higher than the
respective 260/280 values. They are commonly in the range of 1.8-2.2. If the ratio is
appreciably lower, this may indicate the presence of co-purified contaminants.
Baseline correction－If select the baseline correction, the default correction wavelength is
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Chapter 5 Software Operation
340nm. The user can input different wavelength correction according to measurement
requirements. In any measurement, the baseline is automatically set to the absorbance
value of selectable wavelength. All readings under wavelengths should be minus this
value.
Note: The user may elect to turn off the baseline correction, which may result in the
spectra being offset from the baseline.
3.2 Nucleic Acids concentration application steps
1) In the main menu, select nucleic acid mode. It enter into the Nucleic Acids
measurement interface.
2) The users select the type of Nucleic Acid. The user can select 'DNA-50', 'RNA-40',
'ssDNA-33' and 'Other'. The default is DNA-50.
3) In the specific location, input sample name and item number.
4) Create a blanking by using suitable solution. Blank solution，is normally solvent to
dissolve the targeting molecular and it need to be same with the samples in PH and
ionic strength. Load a 1-2ul blank sample onto the lower measurement pedestal and
lower the sampling arm into the 'down' position. Click 'Blank' button to make a blank.
When the correction is complete, the intensity curve will come out. Wipe the sample
from both the upper and lower pedestals using soft laboratory wipe and measure the
next solution.
5) Load a 1-2ul ‘standby’ sample onto the lower measurement pedestal and lower the
sampling arm into the 'down' position. When the correction is complete, the absorbance
curve will come out. In the meantime, the data for this measurement will show in the
measurement result area. The default concentration unit is ng/ul. The default standard
wavelength is 340nm. If the users want to choose a new standard wavelength, enter
into Debug mode. In the meantime, the measurement results will be added into the
‘measurement result list’ in the “List”. If the users want to see more detail absorbance
intensity information, they can see them in ‘Show Detail’ in “List”. The absorbance
intensity data can be stored in Excel file by clicking ‘To Excel’. When click ‘My data’
button to open the file, the measurement result and absorbance curve will be restored
in the software.
6) When the measurement is complete, a new laboratory wipe should be used to wipe the
pedestals. In this way, the users can do next measurement. If measure the same lot
samples, the users do not need to re-blank. It is advisable to do the blanking every
15min at least.
7) The software automatically adds the measurement results into the measurement
results list in “List”. It is best to click 'save' button to save them in the user-specified
region before closing the software, since the data will disappear when closing the
software.
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Chapter 5 Software Operation
Fig 1.Nucleic Acids Measurement Result
Fig2. Measurement Results List
4. Protein A280
4.1 General Information
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Chapter 5 Software Operation
Proteins, unlike nucleic acids, can exhibit considerable diversity. Protein A280 method is
applicable to purified proteins (includes Trp, Tyr residues or Cys-Cys disulfide) exhibiting
absorbance at 280nm. It does not require gereration of a standard curve. The software
calculate the protein concentration directly after measure the absorbance value. These
methods for measure the colors like BCA, Pierce 660nm, Bradford and Lowry, are usually
applicable for the samples with uncertain extinction coefficient or cell Iysate.
The Protein A280 displays UV spectrum, measures the protein’s absorbance at 280nm and
calculate the concentration (mg/ml). Like the Nucleic Acide mode, it displays and records
10mm equivalent data.
Measurement Concentration Range
The Nano-100 Spectrophotometer will accurately measure protein samples up to 100mg/ml
BSA) with dilution. When the optical intensity (after measurement sample extinction) is lower
than 200(under 1cm path length), software will inform the customer to choose shorter path
length to make sure the precision of the measurement. Unique screen is shown as below.
The hydrophobic between the water molecules is the main factor of surface tension. In
general, the presence solute of liquids (（including protein, DNA,RNA, salt ion, detergent
molecule) can significantly reduce surface tension. Although, for most samples, a 1ul sample
size is enough, a 2ul sample size is recommended for protein measurements that the liquid
column be formed.
Screen display:
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Chapter 5 Software Operation
The data shown in the spectrum image are consistency of the reading value at 10mm path
length.
The right column of spectrum includes following information:
Name－Input the sample name: when sample measurement, the sample name should be
input.
Type－Used to select the type of nucleic acid being measured. The user can select ‘A280’
for A280, ‘BSA’ for BSA, ‘lgG’ for lgG, and 'Lysozyme' for Lysozyme.
ID--Display the serial number for the sample being measured. The user can open the
software to start recording.
A260－Absorbance of the sample at 260nm with 10mm path
A280－Absorbance of the sample at 280nm with 10mm path
260/280－Ratio of sample absorbance at 260nm and 280nm.
260/230－ratio of sample absorbance at 260 and 230nm.
Baseline correction－If select the baseline correction, the default correction wavelength is
340nm. The users can input different wavelength correction
according to measurement requirements. In any measurement, the
baseline is readings under wavelengths should be minus this value.
Note: The user may elect to turn off the baseline correction, which may result in the spectra
being offset from the baseline.
4.2 Protein A280 concentration application steps
1) In the main menu, select Protein A280 mode. It enter into the protein measurement
interface.
2) The users select the type of Protein. The user can select 'A280', 'BSA', 'lgG' and 'Lysozyme'.
The default is A280.
3) In the specific location, input sample name and item number.
4) Create a blanking by using suitable solution. Blank solution，is normally solvent to dissolve
the targeting molecular and it need to be same with the samples in PH and ionic strength.
Load a 1-2ul blank sample onto the lower measurement pedestal and lower the sampling
arm into the 'down' position. Click 'Blank' button to make a blank. When the correction is
complete, the intensity curve will come out. Wipe the sample from both the upper and lower
pedestals using soft laboratory wipe and measure the next solution.
5) Load a 1-2ul ‘standby’ sample onto the lower measurement pedestal and lower the
sampling arm into the 'down' position. When the correction is complete, the absorbance
curve will come out. In the meantime, the data for this measurement will show in the
measurement result area. The default concentration unit is mg/ml. The default standard
wavelength is 340nm. If the users want to choose a new standard wavelength, enter into
Debug mode. In the meantime, the measurement results will be added into the
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Chapter 5 Software Operation
‘measurement result list’ in the “List”. If the users want to see more detail absorbance
intensity information, they can see them in ‘Show Detail’ in “List”. The absorbance intensity
data can be stored in Excel file by clicking ‘To Excel’. When click ‘My data’ button to open
the file, the measurement result and absorbance curve will be restored in the software.
6) When the measurement is complete, a new laboratory wipe should be used to wipe the
pedestals. In this way, the users can do next measurement. If measure the same lot
samples, the users do not need to re-blank. It is advisable to do the blanking every 15min
at least.
7) The software automatically adds the measurement results into the measurement results list
in “List”. It is best to click 'save' button to save them in the user-specified region before
closing the software, since the data will disappear when closing the software.
5. Debug Mode
In the Task bar, click "Options" or choose "DebugEnable" item from "Debug", then the
software enter into Debug mode. In that time, the setting keys for user references will be
added in the software interface. It includes:
1) Debug menu
Dropdown menu of Debug incudes:
DebugEnable: select if enter into Debug mode or not.
LampEnable: select if open Asynchronous switch lamp.
R_EliminateBlackI: select if eliminate black current
ContinueBlank: select if automatically continue blanking without clicking.
ContinueMeasure: select if automatically cycle measure without clicking.
SaveBlankData: save the blank data for this measurement
2) Debug tool bar
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The user can set references to satisfy specific measurement requires. If there is any question,
or you have any other requirement for the reference key, you can contact the manufacturer.
Inside the Debug tool bar includes:
Integration time: the integration time for CCD sensor test. The shortest time is not less than
25min.
iSampleNum: the average value for CCD sensor data
Average Num: the average value
Boxcar width: do the smooth processing for absorbance curve of specific wavelength
Boxcar Num: the times to do smooth processing for absorbance curve.
Note: Above set data will be active after clicking "Input"
3) Baseline correction checkbox
Inside the baseline correction checkbox includes：
Baseline correction: select if do baseline correction
Select wave: select the correction wave
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