Azure Ao Absorbance User manual

Azure Ao Absorbance
Microplate Reader
User Manual
Rev 20201009

Azure Ao Absorbance Microplate Reader User Manual Page 2
Foreword
Thank you for purchasing the Azure Ao Microplate Reader. This user manual describes how the instrument
works and how it should be operated. Please read carefully before operating. Keep this user manual for
reference in case you encounter problems.
Opening Inspection
After opening, please inspect the instrument as well as all the accessories that are listed on the packing list.
If there is a mistake, or anything is missing, please contact the distributor or manufacturer.
File No: DCAR0001
Version No: Ao100920

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Safety Warning and Guidelines
Important Information for Safe Use
Read user manual carefully. Users should understand how to use this instrument before operating.
Do not use instrument without reading user manual. Improper operation may result in injury or electric
shock. Read manual carefully and operate using the guidelines described here.
Instrument is for scientic research purposes only!
Security
The operation, maintenance and repair of the Instrument should comply with the basic guidelines and
specications below. Any usage outside of specications may affect the life of the Instrument and the
protection provided.
This product is an indoor Instrument which conforms to Standard B style- I type- GB9706.1.
Protective ground terminal
The ground terminal, intended for connection to an external protective conductor for protection
against electric shock in case of a fault, is located on the inside of the back panel.
Note: There is potential for biological contamination. All biological samples for testing, quality control,
and calibration are regarded as infectious, as well as any part that comes in contact with those
samples. Wear gloves when operating this device.
Before using the device, read manual carefully. This unit is designed for use in laboratory
environments. Device should be used by skilled personnel with appropriate training.
Notice: To avoid injury, keep body parts at least 15 cm away from instrument when it is running.
Operator should not attempt to open or repair instrument. Accidents or loss of warranty may occur. If
something is wrong with the Instrument, please contact the manufacturer for repair.
Before powering on, ensure that the correct voltage and electrical outlet is used (and not lower than
that required). If necessary, the electrical cord can be replaced with the same type. Do not place
objects on top of electric cord.
Hold the jack when pulling out plug, and never pull directly on electrical cord.
When storing instrument, avoid high temperatures, dust, water and UV light. Storage should
include adequate ventilation and instrument should be kept away from corrosive gases, strong
magnetic elds, and heat sources. Avoid obstructing the vent on the instrument as it is designed to
provide aeration.
When nished with the instrument, shut the unit off. Unplug when not in regular use and cover to
prevent dust accumulation.
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Immediately pull the plug from the jack in the following cases, and contact the vendor:
• Fire or ood
• Liquid owing into the instrument
• Signs of abnormal operation (such as sound or smell)
• Instrument is dropped or outer shell damaged
• The function has obviously changed
The Maintenance of Instrument
Microplate holder and instrument outer shell can be cleaned using a cloth dampened with alcohol.
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Table of Contents
1. Brief Introduction 6
2. Features 7
2.1 Working Conditions 7
2.2 The Basic Parameters and Characteristics 7
3. Installation 9
3.1 Initial Inspection 9
3.2 System Placement 9
3.3 Installation Steps 9
4. Operation Guide 10
4.1 Instrument Self Check 10
4.2 Ao Software Overview 10
4.3 The Results Tab 11
4.4 The Settings Tab 12
4.5 The Help Tab 12
5. Operation Quick Guide 13
6. Detailed Software Operation 14
6.1 Creating a New Protocol 14
6.1.1 Measure 14
6.1.2 Layout 16
6.1.3 Shake Setting 19
6.1.4 Calculate 20
6.1.5 Interpret 23
6.1.6 Quality Controls 23
6.2 Report 25
6.2.1 Ordinary Report Interface 25
6.2.2 Kinetics Report Interface 27
6.3 Records 27
6.4 Settings 29
6.4.1 Language Setting 30
6.4.2 Filter Setting 31
6.4.3 Maintenance 32
6.4.4 Time and Date setting 32
7. Maintenance, Storage & Transportation 34
7.1 Maintenance 34
7.1.1 Light Source Replacement 35
7.1.2 Add Optical Filter 35
7.2 Storage and transportation 36
8. Troubleshooting 39

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1. Brief Introduction
The Azure Ao Absorbance Microplate Reader is a professional instrument for performing EIA tests, measuring
concentrations, measuring absorbance, and performing Enzyme - Linked Immunosorbent Assays (ELISA).
This type of reader is widely used in biology, agriculture, food and environment research, and is greatly
benecial to the growing number of ELISA kit users.
Highlights
1. Easy to use, with 7 inch touch screen and 3 external keys.
2. English/Chinese operation system can edit, measure, and save data automatically.
3. Stand-alone instrument does not require an external computer for operation.
4. 8-position optical lter wheel, equipped with 5 standard lters and 3 custom lters.
5. 9-channel vertical optical path, zero dispersion single mode ber measurement system, automatic plate
well center position function.
6. 96-well visual layout helps to label the blank, samples, positive/negative controls, quality controls and
multi-value control.
7. Perform multiple tests on a single plate.
8. Single or double wavelength measurement.
9. Self-checking optical path, mechanical motion.
10. Plate-shaker function, with adjustable time and speed.
11. Designed with energy-saving light source.
12. Multiple data export ports.
13. Measurement results can be exported to MS Excel le.

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2. Features
2.1—Working Conditions
• Ambient temperature: 41°F – 95°F (5°C – 35°C)
• The relative humidity: ≤70%
• Power: AC100-240V 50-60Hz 2A
2.2—The Basic Parameters and Characteristics
Parameter/Model Ao Absorbance Microplate Reader
Light Source 6V10W Quartz-Halogen lamp
Wavelength 340nm – 750nm
Optical Filter Five standard lters – 405, 450, 492, 595 and 630nm with a max load of 8 lters
Read-out Range 0.000 – 4.000Abs
Resolution 0.001Abs
Linearity CV≤±1%(0.000 – 3.000A) , CV ≤±2%(3.000 – 4.000A)
Precision CV≤0.3%(0 – 3Abs); CV≤2%(3 – 4Abs)
Accuracy (450nm) ±0.005 or ±1%(0 – 3Abs), ±2%(3 – 4Abs)
Channel Deviation <0.01A—450nm 1.0Abs
Speed Single wavelength <15s/96well, double wavelength<28s/96well (Normal mode)
Dimension (WXDXH) 440x295x225mm
Weight (kg) 10kg

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• Plate holder: The plate holder holds the microplate securely within the drawer. Align the A1 of the plate
to the designated marking within the plate holder.
• USB port: Use USB ports to export data from the instrument or to connect a printer, keyboard or mouse.
• Back door: Access the light source or optical lter wheel.
• Knob: Use the knob to open the back door of the instrument.

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3.1—Initial Inspection
Each Ao Microplate Reader is inspected before it is packaged and shipped. Upon receipt, please perform
an initial inspection to conrm all parts are received and in good order. Contact your local distributor or
manufacturer in case:
• The outer package has evidence of moisture
• The outer package has evidence of damage
• The outer package has been opened or tampered with
If the packaging is intact, check the instrument and packing list.
• Check all accessories according to the packing list
• Inspect instrument appearance for cracks, warping, or damage.
3.2—System Placement
• Place instrument on a at, dry, clean surface ensuring enough space in the front for the plate drawer
to extend.
• Ensure at least 15 cm of space on each side and behind the instrument.
• Working environment:
• Atmosphere is free from corrosive steam or smoke.
• Temperature within +10°C to +40°C.
• Make sure there is no condensation or high humidity (Relative humidity from 10% to 80%).
Note: KEEP INSTRUMENT AWAY FROM CORROSIVE GAS OR LIQUID!
3.3—Installation Steps
1. Place instrument on work bench.
WARNING: Do not remove any screws or parts unless directed by manufacturer or distributor.
Doing so may void warranty.
2. Connect instrument to power supply.
3. Instrument will start self-testing after powering on and will be immediately ready for use.
Note: Do not connect instrument to an ungrounded outlet.
3. Installation

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This chapter introduces default software operation.
4.1—Instrument Self Check
When powered on, the software immediately performs a self-check to verify operation of light source and
lters. After completion of the self-check, instrument will enter the main software interface. See Fig 1.
4.2—Ao Software Overview
The Azure Ao operates with an intuitive, touch-based user interface. All assay parameters and instrument
functions can be accessed through four tabs at the top of the main interface – Protocol, Results, Settings,
and Help.
4. Operation Guide
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The following can be accessed from the Protocol Tab
1. Active Protocol – The name of the active protocol will be displayed here.
2. File List – Access, export or upload previously saved protocols and data les.
3. Save – Save changes to the currently active protocol.
4. Save As – Save changes to the currently active protocol as a new protocol.
5. New – Create a new protocol.
6. Measure – Select desired lter(s) and read speed (fast or normal mode).
7. Layout – Assign wells as standards, controls, unknowns, blanks or empty. If using standards, tap
Concentration to enter the standard values.
8. Shake – Control the plate shaking function – activate shake function, speed and time.
9. Calculate – Control pre-processing of multiple wavelengths, curve ts and kinetic reading parameters.
10. Interpret – Set algorithm for determining positive unknown samples based on positive and negative
control values.
11. Quality – Enter Quality Control acceptable ranges in order to determine assay validity.
12. Start – Start plate read.
13. Stop – Stop plate read.
14. Plate In/Out – Open and close the plate drawer.

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4.3—The Results Tab
After a plate is read, the software will automatically open in the Results Tab. The Results tab allows users to
view and export raw data, analyzed data, samples interpreted as positive or negative, the standard curve,
and results of the quality control analysis if set.
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1. Raw Data – View and export raw optical density data. If a blank is set, the average blank value will be
displayed on the right under Filter.
2. Interpret – If values were set in the Interpret algorithm, click this tab to view the results. A red “+” symbol
indicates a strong positive sample. A yellow “+” symbol indicates a weak positive.
3. Analysis – If a standard curve was set, the results interpolated from the standard curve will be displayed
and can be exported from here.
4. Curve – View and export the standard curve. The curve t can be adjusted from this screen as well.
5. QC – If QC acceptance limits were entered, the results of all QC samples will be displayed here along
with indications of pass or fail.
6. Filter – The lter used to read the plate is displayed here. If multiple lters were used, touch this button to
toggle between the raw data for each lter or the pre-processed data if pre-processing was used.
7. Export – Click to export data to USB.
8. Print – Click to print using an Ao Printer.

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4.4—The Settings Tab
Basic instrument Settings can be accessed through the Settings tab.
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1. Language – Set instrument language to English or Chinese. Language is set to English my default.
2. Filter – View or adjust the lter positions on the lter wheel.
Note: Positions must math the physical positions on the lter wheel.
3. Print – Turn on and set report titles for printing.
4. Date & Time – Set the date and time of the instrument.
5. Maintenance – The following can be accessed from the
Maintenance tab:
• Plate – Set the default plate drawer position to In or Out
• Reset – Reset the instrument to manufacturer settings.
• Raw Intensity – Set instrument to return Optical Density or Raw light intensity measurements.
Note: Other settings in this tab are password protected. Azure Biosystems does not recommend
accessing these settings as they are reserved for use during the instrument manufacture process.
4.5—The Help Tab
View basic usage instructions, software version information, and Azure Biosystems contact information.
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1. Turn the instrument on by ipping the switch on the back right corner of the instrument.
• The instrument will go through a self-check before entering the main Program interface.
2. To run an existing protocol press File List to access saved protocols and select the desired protocol.
3. To create a new protocol press New and enter a new le name to create a new protocol.
4. Set or verify the protocol parameters
• Measure – Set or verify the desired lters and read speed.
• Layout – Set plate layout by assigning well types
1. BK – Blank well
2. SD – Standard. Double tap to select different standards. Enter standard values and units in the
Concentration window.
3. QC – Quality Control. Double tap to select different QCs.
4. UD – Unknown
5. EP – Empty Well
6. NC – Negative Control
7. PC – Positive Control
5. Activate or adjust the plate Shake function.
6. Set or verify curve t, wavelength pre-processing, or kinetic function from the Calculate menu.
7. Press Plate In/Out to open the plate drawer.
• Align well A1 of the 96-well plate with the A1 designation in the plate drawer.
• Gently angle the plate toward the metal plate tension piece and press plate against the tension piece
until it gives enough to slide the plate into place.
8. Press Start to read the plate. If the protocol was not saved, the instrument will prompt user to enter a le
name. The protocol and data will be saved as a single le.
9. After the plate is read, the software will automatically move to the Results tab. View, print or export raw
data or analyzed data from this tab.
5. Operation Quick Guide

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The main interface offers access to all assay parameters and instrument function as well as access to the
four main tabs: Program, Results, Settings, Help.
6.1—Creating a New Protocol
1. Create a new program.
Click “New”, input a new program name, click “OK” or “return”. and Help.
2. Save current program.
After creating a new program and setting parameters, hit “Save” to save all settings. After reading a
plate, hit “Save” to save plate data to the current program le.
Note: Once a program has been used to read a plate and data has been saved to the le, some
parameters of that program will be locked and grayed out. Program parameters may only be changed
before the program has been used to read a plate and data has been saved.
3. Save as new program.
To use a saved program as the base of a new program, open the saved program and tap “Save As.”
Give the program a unique name and hit “Return.” You may now change any parameters desired before
reading a plate.
“Save As” can also be used after a plate has been read to save the plate data and program parameters
to a new le. After reading a plate, hit “Save As”, change the le name and hit “Return.”
4. Open a saved data le.
Touch “File List” to retrieve saved program les. If the le was saved after reading a plate, selecting the
saved le will open the program along with the saved plate data and results reading a plate.
6. Detailed Software Operation
Figure 4

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Figure 5
6.1.1—Measure
Wavelength and mode (read speed) can be set in measurement mode.
1. On the “Protocol” interface, touch “Measure” to select up to two wavelength lters and to set the
measurement speed. See Fig 5.
2. Filter1 Setting
Touch the selection box to the right of “Filter 1” to select a lter. See Fig 6.
There are 5 standard lters included: 405nm, 450nm, 492nm, 595nm, and 630nm – as well as 3
additional customer selected lters. Choose one as Filter1.
Figure 6

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Figure 7
3. Filter2 Setting
The instrument can perform single or dual wavelength measurements. To select a second lter, touch
the selection box next to “Filter 2” and select the desired lter. Select “No Filter” to perform a single
wavelength measurement. See Fig 7.
Note: For single wavelength measurement, choose “NO Filter” for Filter2.
4. Measurement speed
There are two options: Fast or Normal. Touch the selection box to the right of “Mode” to choose the
desired read speed.
Note: “Normal” mode is recommended.
5. Press ”OK” to nish setting and return back to main interface.
6.1.2—Layout
Plate Layout
1. In the “Protocol” interface, you can touch the box to the right of “Plate Layout” to bring up the Plate
Layout Interface. See Fig 8.
Figure 8

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2. To set a plate layout:
First, choose the sample type (the active type will be marked with a red frame, like the UD (unknown
sample) type in the gure above. Next, select well positions on plate layout. The selected positions on
the layout will change to the color of the sample type.
3. Choose a Type from these seven: BK,SD,QC,UD,EP,NC,PC dened below:
Blank well for blank control. You can set several wells as blank.
Standard sample well for setting the standard curve. Press (see Fig 9) to select between
different sample numbers. Select the desired standard to activate it, then select the appropriate well
position on layout.
Figure 9
Set a maximum of 12 standard samples. Several wells can be set with the same number.
QC control. Steps for setting up the QC control samples are the same as above for setting
standards. (See Fig 10).
Figure 10
You can set up to 9 QC controls and can set several wells with the same number.

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Unknown sample. Set several positions as unknown samples. Instrument’s default setting is for
all positions on plate to be set as unknown samples.
Empty wells. Select EP to designate empty wells; the measurement result will show EP instead
of data.
Negative control. It is possible to set several positions as negative controls.
Positive control. It is possible to set several positions as positive controls.
4. After setting all sample types in their respective positions, press “OK” to return to the “Programs”
interface and start the measurement. If there are standards, input the standard sample concentration
and then begin measurement.
Note: You can load the microplate and then set the layout type or vice versa.
Standard sample concentration setting
From the “Plate layout” interface, press “Concentration” on the right. This will bring up the concentration
interface shown below (see Fig 11).
Figure 11
1. The interface will show all 12 standard samples, but you only need to set the concentration for the
positions which have been selected on the layout. Standards that have not been selected on the
layout will not be measured.
2. When setting concentration, click the corresponding No. to input the concentration value.
Note: Make sure that the No. of each sample position on the layout corresponds with the microplate
and that the sample concentration is correct. Any mistake will affect curve tting and the
measured result.
3. “Unit” can be set by pressing ”Unit” button on the right. See Fig 12.

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Figure 12
4. Press “OK” to conrm and return to main interface.
6.1.3—Shake Setting
This instrument can shake the plate to mix samples and knock drops off of the walls of the well before
measuring to ensure an accurate measurement.
1. On “Programs” interface, click “Shake” button under “Shake parameters” to set shake speed and time.
Figure 13
2. Click button beside “Shake” to choose “On” or “Off”. See Fig 14.
Figure 14

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3. Click button beside “Speed” to set speed: Slow, Medium, or Fast. See Fig 15.
Figure 15
4. Click button beside “Time” to set shake time. Input numbers to set HH/MM/SS. See Fig 16.
Figure 16
5. Press to complete settings and return to “Programs” interface.
6.1.4—Calculate
This section allows you to set the sample result processing method, including Kinetic, Preprocess,
or Curves.
Kinetic setting
1. On “Programs” interface, click box to the right of ”Calculate” to set “Calculate parameters”. Click button
beside “Kinetic” to choose On or Off. Set parameters for kinetic reads when set to “On.” See Fig 17.