Enzo Absorbance 96 User manual

Rev 052021
Product Manual
Absorbance 96 plate reader
Catalog #: ENZ-INS-A96
Manufactured by Byonoy GmbH
Device Version 2.0 (2021-01-18)
Software Version 1.2 (2021-01-18)
For the latest product information, including support documentation, visit us
online:
www.enzolifesciences.com
Technical Support (US): 800-942-0430
Technical Support (EU): [phone]
Customer Service (US): [email protected]
Customer Service (EU): [email protected]
FOR RESEARCH USE ONLY.
NOT FOR USE IN DIAGNOSTIC PROCEDURES.

Manufacturer:
Byonoy GmbH
Bernstorffstr. 118
22767 Hamburg, Germany
00 49 40 5379 866 00
[e-mail]
www.byonoy.com
Instructions for Use
Absorbance 96
Valid for REF Number ABS MH A 01/ABS MH D 01
Instructions for Use
Absorbance 96
Version 2.0 (18/01/2021)
©Copyright 2021 by Byonoy GmbH. All rights reserved.

II
Dear customer,
We are delighted that you have chosen the Absorbance 96. To take full advantage of the
instrument's performance and to enjoy your instrument for many years, please read these
instructions for use carefully before installation and commissioning. Operate the instrument
in accordance with these instructions. The operating-safety and function of the instrument
can only be guaranteed if both the general safety regulations and accident prevention reg-
ulations of the legislator as well as the safety instructions in this manual are observed. We
accept no liability for any damage resulting from improper use or incorrect operation.
Ensure that the manual is always accessible and is read and understood by all
persons operating the instrument.
This user manual may only be used according to its intended purpose. It may not be repro-
duced, changed, or translated in another language without the prior written consent of
Byonoy GmbH.
This document is subject to technical changes and updates.

III
TABLE OF CONTENTS
1 General information...................................................................................................................... 1
2 Overview of the Absorbance 96.......................................................................................... 3
2.1 Scope of application ................................................................................................................................. 3
2.2 Measurement method ............................................................................................................................. 3
2.3 Absorbance 96 System........................................................................................................................... 3
2.4 Consumables .................................................................................................................................................. 5
3 Preparing the product for use..............................................................................................6
3.1 Unpacking, storage, and transport ................................................................................................ 6
3.2 Supplied materials...................................................................................................................................... 6
3.3 Working environment .............................................................................................................................. 6
4 Operation ........................................................................................................................................... 8
4.1 App installation procedure and updates................................................................................... 8
4.2 Computer connection.............................................................................................................................. 8
4.3 Assay readout................................................................................................................................................ 9
5 Signals and troubleshooting................................................................................................ 10
5.1 Signal lights.................................................................................................................................................... 10
5.2 Troubleshooting ........................................................................................................................................ 10
6 Maintenance and cleaning .................................................................................................... 12
6.1 Maintenance ................................................................................................................................................. 12
6.2 Cleaning............................................................................................................................................................ 13
6.3 Technical support..................................................................................................................................... 15
6.4 Repairs .............................................................................................................................................................. 15
7 Technical specifications ......................................................................................................... 16
8 Guarantee ........................................................................................................................................ 18
9 EC-Conformity Declaration................................................................................................... 19

General information
1
1 General information
Important: Follow the instructions for use
Any commissioning or handling of the instrument requires precise knowledge and notice of
this manual. The device is intended for the described use only.
In this manual, particularly important remarks are labeled as followed:
Warning:
This is a warning that indicates an immediately or potentially dangerous situa-
tion, which, if not avoided, will result in death or serious injury.
Caution:
Caution indicates a potentially hazardous situation, which, if not avoided, could
result in minor or moderate injury.
Information:
This is a piece of information indicating certain properties that must be ob-
served.
Liability for function and damage
Liability for the function of the device shall, in any case, pass to the owner or operator if the
device is improperly maintained, repaired, or modified by persons who do not belong to the
authorized service personnel, or if it is handled in a way that does not comply with its in-
tended use. The service and operation of the product must be in accordance with this man-
ual. Byonoy shall not be liable for damages resulting from non-observance of the information
above. Warranty and liability conditions of the terms of sale and delivery of Byonoy are not
extended by the information above.
Warning
An incorrect working environment can lead to a reduction in service life, dam-
age to the device, or measurement errors. Please note the explanations and
warnings in chapter 3.3.
To avoid measurement errors, it is essential to ensure that the instrument is
connected properly and that the experiments are carried out correctly. Please
note the explanations and warnings in chapter 3.
Incorrect cleaning of the instrument can reduce its service life and can cause
damage to the instrument. Please note the explanations and warnings in chap-
ter 6.2.
Information
Follow all safety instructions on the device and in the attached documents.

General information
2
Follow all general precautions that apply to electrical instruments.
Disposal of used electrical and electronic equipment
The symbol on the product or its packaging indicates that this product is not to be treated
as normal household waste. It must be disposed of at a collection point for the recycling of
electrical and electronic equipment. By contributing to the correct disposal of this product,
you protect the environment and the health of your fellow human beings. Recycling helps to
reduce the consumption of raw materials. For further information on how to recycle this
product, please contact your local authority or municipal waste disposal centres.
Working with biological and harmful material
The Absorbance 96 is not to be used for the measurement of biohazardous substances.
Always observe the manufacturer's hazard information pertaining to the substances to be
measured.
The instrument does not produce any toxic or harmful gases or substances. During the
measurement, make sure that there are no toxic or harmful substances in the microtiter plate.

Overview of the Absorbance 96
3
2 Overview of the Absorbance 96
2.1 Scope of application
The Absorbance 96 is an optical laboratory instrument for measuring the absorbance (opti-
cal density) of biological or non-biological samples in ANSI/SBS-standard 96-well microtiter
plates in accordance with the specifications described in the user manual. The Absorbance
96 is intended for research and other non-in-vitro-diagnostic analyses only. It is to be oper-
ated by trained laboratory personnel and is intended for professional use.
2.2 Measurement method
The Absorbance 96 is designed to carry out sensitive absorbance measurements. It
measures the optical density (OD) of samples at defined wavelengths.
Absorption
Absorption refers to the amount of light absorbed by a medium. Absorption reduces trans-
mission. Transmission is the ratio of incident to transmitted light. Accordingly, the degree of
transmission is calculated as follows
T = (I/I0)
where I is transmitted light and I0is incident light.
Optical Density
Optical density is a measure of the attenuation of light radiation after it has passed through
a medium. Optical density is the logarithmic quantity that describes the reciprocal of the
transmittance T:
OD = log(I0/I)
where I is the transmitted light and I0is incident light.
Optical density is the absorbance of the sample plus other attenuating effects such as scat-
tered light due to turbidity. It is therefore necessary to avoid scattered light in order to meas-
ure absorbance correctly.
If no other attenuating effects are present, absorbance = optical density.
2.3 Absorbance 96 System
Absorbance 96 Reader
The Absorbance 96 reader is an absorbance-based measuring device, i. e. a measuring in-
strument, with which the absorbance values of a 96-well microtiter plate can be read, rec-
orded and provided for further processing. The Absorbance 96 is a solid-state microplate
reader and has 96 detection units, allowing measurement without a scanning mechanism.

Overview of the Absorbance 96
4
In the upper part of the measuring instrument, there are up to 4 LED light sources with dif-
ferent wavelengths and up to 4 interference filters. The four signal lights on the top of the
reader show which measuring channel is being used for the current measurement.
The microtiter plate is inserted manually into the slot of the device. There are two raised
locking-edges at the bottom of the slot, as well as an arrow for visual inspection. The arrow
becomes wholly visible only after complete insertion of the microtiter plate into the slot.
Figure 1.
Visual representation of the product from the front, top. 1. Signal lights. 2. Slot for
microtiter plate. 3. Raised locking-edges. 4. Arrow.
On the back of the Absorbance 96 there is a Micro-USB port for connecting the instrument
to a computer via the included Micro-USB cable. Furthermore, there is an eject button for
ejecting the microtiter plate out of the slot.
Figure 2.
Visual representation of the product from behind, top. 5. Eject button. 6. Micro-
USB port.
Power consumption
The Absorbance 96 is an analytical instrument with very low power consumption. It is oper-
ated by a Micro-USB cable and the total power consumption is generally less than 2.5 watts.
Absorbance 96 App

Overview of the Absorbance 96
5
The reader is controlled via the Absorbance 96 App. The operation of the app is described
in-detail in a separate user manual.
2.4 Consumables
With the Absorbance 96, ANSI/SBS Standard 96-well microtiter plates can be used. Only
certain microplate types can be used with the Absorbance 96 (see chapter 7).
When using microtiter plates, always check the specifications of the microtiter plate manu-
facturer. Not all microtiter plates of a particular manufacturer are the same in design, mate-
rials, or configuration. The temperature stability within the microtiter plate can depend on
the type of microtiter plate used.

Preparing the product for use
6
3 Preparing the product for use
3.1 Unpacking, storage, and transport
Unpacking
Remove the packaging material and carefully place the device on a firm and level surface.
Check the device for external damage and check the materials supplied (see chapter 3.2).
The device is packed in a specially designed cardboard box. Keep the packaging material. If
the device must be returned for repair, the original packaging material must be used.
Storage and protection during the intervals of normal use
Protect the device from moisture and dust during prolonged storage.
Do not store the device next to heat sources and protect it from direct sunlight.
The temperature for storage should be within the recommended temperature range (see
chapter 7).
Transportation
Before transportation, unplug the instrument and ensure there is no microtiter plate inside
the device. Depending on the transport distance, use the original packaging material. Make
sure that the new location meets the requirements described in chapter 3.3.
To avoid spillage and contamination, make sure that there is no microtiter plate
inside the instrument before transport.
Always transport the product well-packaged to avoid damage in transit.
3.2 Supplied materials
When unpacking the device, please check that the following components are present:
• Absorbance 96
• Micro-USB cable
• Hex key
• Microfiber cloth
• USB Stick
3.3 Working environment
Due to its small size, the Absorbance 96 can be operated very well in various working envi-
ronments.

Preparing the product for use
7
However, the following warnings must be observed. Ignoring them may lead to measurement
errors and a reduction in the expected lifetime or damage to the device:
The device should stand on a level surface, free from dust and vibrations.
Do not operate the device near heat sources or under direct sunlight.
The ambient temperature and the humidity should be within the recommended
range (see chapter 7).
Place the instrument in a horizontal position secured against falling.

Operation
8
4 Operation
Before commissioning the Absorbance 96, you should carefully read and understand the en-
tire manual to familiarize yourself with the system.
The Absorbance 96 is controlled via the Absorbance 96 App, including the analysis of meas-
urement data. The functions of the app are detailed in a separate user manual.
4.1 App installation procedure and updates
The app is compatible with Windows und MacOS operating systems (see chapter 7).
To install the Absorbance 96 App, insert the USB stick into the computer and follow the steps
below:
• Locate and select the USB drive in the document browser.
• Open the installation file either for Windows or macOS.
• Follow the steps in the Setup Wizard to complete the installation.
Modern operating systems offer plug and play functionality. Older operating systems may
require manual installation of the software drivers. The drivers for Windows are available on
the USB stick.
To install the proper drivers and software on the computer, the user must have
administrator rights or obtain such permissions.
4.2 Computer connection
The Absorbance 96 requires a connection to the computer for power supply, starting the
experiment, and data transfer. Insert the Micro-USB cable into the Absorbance 96 and the
USB port of the computer. The Absorbance 96 starts automatically when connected to a
computer via the Micro-USB cable and the software automatically establishes a connection.
Always use the included Micro-USB cable for the connection to your computer.
Connect the instrument directly to your computer and do not use an external
USB hub.
The device may only be operated at a USB port USB 2/USB 3 with 5 VDC and a
maximum of 3 A of a certificated computer (with certification mark of an ap-
proved testing laboratory). Use an adapter cable on USB 3.1 with type C plug
connection, which ensures the profile 1: 5 [email protected] A.
When the Absorbance 96 is switched on, the signal lights on the top of the instrument light
up. Each time the instrument is switched on, an internal self-test is performed to ensure there
are no malfunctions.
If no connection between the instrument and the computer is established, install the driver
for the USB connection. You can find it in the supplied USB Stick.

Operation
9
4.3 Assay readout
Once all settings have been completed, the measurement can be started. The app will auto-
matically guide the user through the readout process.
In order to guarantee a correct measurement result, please pay attention to the indications
in the Absorbance 96 App und heed the following warnings:
When inserting the microtiter plate, pay attention to its alignment.
Make sure that the microtiter plate was inserted in the slot completely. In that
case, the arrow at the bottom of the slot is visible completely.
The bottom of the microtiter plate must be dry before it is inserted into the in-
strument. If the microtiter plate is wet at the bottom, dry it with a paper towel
or similar.
Shocks of the Absorbance 96 or the microtiter plate can cause an unwanted
signal. Therefore, neither the Absorbance 96 nor the microtiter plate should be
touched during measurement.
Note that the system depends on a clean and dust-free environment. There-
fore, do not wear powdered safety gloves.
Ensure that no direct sunlight or other strong light sources shine into the slot
of the Absorbance 96.
Insert the plate cautiously to avoid spills.
After a successful measurement, the results section will open automatically in the app.
The microtiter plate may now be removed from the Absorbance 96. To do this, push the
eject button on the rear side of the device.

Signals and troubleshooting
10
5 Signals and troubleshooting
5.1 Signal lights
Table 1.
Explanation of the signal lights
Activity of the signal light
Meaning
All signal lights flash simultaneously
Error, software shows error message
No signal lights light up
Device is off
Device is waiting for input
Device is defective
All signal lights light up briefly in sequence
Self-test after connection to power
Signal light of the respective measuring
channel flashes/illuminates continuously
Initialization/measurement in progress
5.2 Troubleshooting
If there is an error, all four signal lights flash simultaneously. In this case, check the software
to see which error is present.
Table 2.
Error messages and solutions
Type of error
Cause
Solution
Initialization failed.
There was a microtiter
plate in the slot during ini-
tialization.
Remove the plate.
The inside of the slot is
dirty.
Clean the inside of the slot ac-
cording to the cleaning instruc-
tions in chapter 6.2.
Irrecoverable hardware
error.
Contact the service department.
Interference due to
ambient light that is
too bright.
Too much ambient light is
entering the slot.
Make sure the instrument is not
exposed to too much ambient
light, e. g. direct sunlight, during
the measurement process.
Temperature out of
specification.
The ambient temperature
was too high or too low.
Make sure the ambient tempera-
ture is within the specifications.
USB Power Supply
Defective (< 450mA or
< 4V).
Defective USB port.
Use a different USB port on the
computer or a different computer.
Using a USB hub.
Connect the instrument directly to
the computer or use another USB
hub.
Defective Micro-USB ca-
ble.
Contact the service department.

Signals and troubleshooting
11
Irrecoverable hard-
ware error.
Irrecoverable hardware
error.
Contact the service department.

Maintenance and cleaning
12
6 Maintenance and cleaning
6.1 Maintenance
The Absorbance 96 is maintenance-free. Each time the instrument is switched on, an internal
self-test is carried out to ensure there are no malfunctions.
There are no parts within the Absorbance 96 that can be serviced by the customer. It is only
necessary to ensure that the device is kept clean.
The accuracy, linearity and reproducibility of the device can be checked with a reference
plate. Byonoy recommends the reference plate from Hellma GmbH (item code 666-R013), for
which the following steps are described.
Accuracy
1. Measure the plate at the desired wavelength. Ensure that there is no dust or other
contamination on the reference plate.
2. Compare results taking the measurement uncertainty and specifications of the
reader into account and use the following formula:
ODRP - MU - AS ≤ODRES ≤ODRP + MU + AS
ODRP:Optical density of the reference plate at the desired wavelength. This value is
found on the calibration certificate from the manufacturer of the reference plate.
MU: Measurement uncertainty of the reference plate. This value is found on the cali-
bration certificate from the manufacturer of the reference plate.
AS: Accuracy specification of the reader at measured optical density. This value is
shown in chapter 7.
ODRES: The optical density measured by the reader at the desired wavelength.
Example:
ODRP:1.554
MU: ± 0.0079
AS: ≤1 % + 0.010 OD from 0.0–2.0 OD
ODRES: 1.573
(1) 1.554 - 0.0079 - (1.554*0.001 + 0.010) ≤ODRES ≤1.554 + 0.0079 + (1.554*0.001 +
0.010)
(2) 1.52056 ≤ODRES ≤1.58744
The measured optical density of the reader lies within the calculated range.
3. Rotate the reference plate by 180 degrees and repeat steps 1 and 2.
4. If all Results (ODRES) are within the range, the accuracy of the device has been veri-
fied.

Maintenance and cleaning
13
Linearity
With the Hellma reference plate, you can check the accuracy at 6 different OD points. If the
accuracy for all these points has been verified, the linearity of the reader is also verified.
Reproducibility
1. Measure the plate at the desired wavelength for at least 3 times. Ensure that there is
no dust or other contamination on the reference plate.
2. Calculate the standard deviation ODSD for each well.
3. Compare the measured standard deviation and specifications of the reader using the
following formula:
ODSD ≤LS
ODSD: Calculated standard deviation for each well
LS: Linearity specification of the reader at measured optical density. This value is
found in chapter 7.
In case that the results of accuracy, linearity or reproducibility are outside the range, please
contact the service department.
Please follow the plate manufacturer’s recalibration instructions for the refer-
ence plate.
6.2 Cleaning
Remove the Micro-USB cable from the instrument before cleaning.
Make sure before cleaning, that there is no microtiter plate in the instrument.
Cleaning the housing
The surfaces of the housing should be cleaned regularly. For this purpose, you can use e. g.
a cloth, or a sponge lightly wetted in water. For heavier soiling, clean the surface of the hous-
ing with a mild soap solution diluted with water or glass cleaner and then wipe with a lightly
moistened cloth or sponge to remove any residue. Do not use scouring agents.
Cleaning the slot
To clean the slot of the Absorbance 96 open the four screws on the bottom of the instrument
(see Figure 3) using the Hex key provided.
Do not loosen any screws that are not explicitly marked in the manual, as this
can lead to a malfunction and may void the instrument's warranty.

Maintenance and cleaning
14
Figure 3.
Bottom side of the instrument
Figure 4.
Pull the instrument apart into its two components
Carefully pull apart the top and the bottom of the instrument to separate the plug connection
on the back of the instrument.
To avoid damaging the rigid plug connection between the top and bottom of
the instrument, grasp the instrument at the back during the pull movement and
avoid any tilting movement.
Use a dry microfiber cloth or a bellows to carefully remove dust or dirt from the surface. If
this is not enough, the microfiber cloth can be slightly soaked with 80% ethanol to carefully
remove dust or contamination from the surface.

Maintenance and cleaning
15
Do not allow water or other liquids to get inside the instrument. If this happens, return the
device to the manufacturer for inspection.
Warnings for cleaning
When cleaning, pay attention to the following warnings. Failure to observe these warnings
may result in damage to the instrument. This can lead to a reduction of the service life or in
measurement errors:
Always avoid spraying liquid directly onto the surfaces of the Absorbance 96.
This is especially important for the inside of the slot, where there are optical el-
ements that are very sensitive and essential for the functioning of the instru-
ment.
Never clean the inside of the slot with sharp or abrasive scourges, and do not
use aggressive solvents or corrosive agents.
Biological hazard: Always wear gloves during cleaning operations that may in-
volve contact with biological or generally hazardous materials or liquids.
6.3 Technical support
In case of problems with the instrument, please contact the manufacturer’s service depart-
ment or your local representative.
Biological hazard: It is your responsibility to decontaminate the instrument and
all accessories before servicing and before returning the instrument or acces-
sories to the manufacturer.
For decontamination of the Absorbance 96, follow the government guidelines for inactiva-
tion of organisms used in biological laboratories.
6.4 Repairs
Repairs on the device may only be carried out by the manufacturer. Please contact the ser-
vice department. The product warranty is voided if the device is modified by unauthorized
persons, or different parts are installed.

Technical specifications
16
7 Technical specifications
Table 3.
Performance and technical data
Parameter
Value
Type of product
Absorbance reader
Product name
Absorbance 96
Service life
10 years at an average use of 4h/day
Housing material
Aluminium
Place for use
Laboratory
Degree of contamination
2
Temperature for storage/measurement
5–40 °C
Temperature (Transport)
-10–50 °C
Relative tolerated humidity (storage/measure-
ment)
Max. 70 %
Measurement method
Absorbance
Measuring technique
Endpoint and Kinetic
Microplate types
96-well, flat bottom
Light source
Up to 4 LEDs
Filters
Up to 4
Detection
96 Photodiodes
Measurement range
0–4.0 OD
Resolution
0.001 OD
Accuracy*
405 nm:
≤1.5 % + 0.010 OD from 0.0–2.0 OD
≤3 % + 0.010 OD from 2.0–3.0 OD
≥450 nm:
≤1 % + 0.010 OD from 0.0–2.0 OD
≤1.5 % + 0.010 OD from 2.0–3.0 OD
Reproducibility**
≤0.5 % + 0.005 OD from 0.0–2.0 OD
≤1 % + 0.010 OD from 2.0–3.0 OD
Linearity***
405 nm:
≤1.5 % from 0,0–2.0 OD!
≤3 % from 2.0–3.0 OD
≥450 nm:
≤1 % from 0,0–2.0 OD
≤1.5 % from 2.0–3.0 OD
Connection to computer
USB 2/USB 3 with 5 VDC and max. 3 A
Dimensions
55 x 96 x 154 mm
Power supply
5 VDC