2010 - 2020 Microplate Absorbance Reader 2020 Sw-Version 2.0 Up Operating Manual Ver 03 - 10
2010 - 2020 Microplate Absorbance Reader 2020 Sw-Version 2.0 Up Operating Manual Ver 03 - 10
2010 - 2020 Microplate Absorbance Reader 2020 Sw-Version 2.0 Up Operating Manual Ver 03 - 10
Operating Manual
manual version: 03/10
Biochrom Ltd
22 Cambridge Science Park
Cambridge
UK
CB4 0FJ
E-Mail: enquiries@biochrom.co.uk
anthos 2010/2020 Operating Manual
Table of Contents:
Safety Information. 4
1 Introduction..........................................................................................................7
1.1 Intended use ..................................................................................................... 7
2 Technical Information .........................................................................................8
2.1 General Information regarding the Absorption Photometer................................ 8
2.2 Metrological specification .................................................................................. 8
2.2.1 Measuring system ............................................................................................................ 8
2.2.2 Measuring process ........................................................................................................... 8
2.2.3 Measuring range, specified according to DIN 58960 ....................................................... 8
2.2.4 Rated operating conditions .............................................................................................. 8
2.2.5 Limits of error and tolerance limits ................................................................................... 9
2.3 General technical data .................................................................................... 10
2.3.1 Weight and Dimensions ................................................................................................. 10
2.3.2 Environmental influences ............................................................................................... 10
2.3.3 Electrical Rating ............................................................................................................. 10
2.3.4 Interfacing ....................................................................................................................... 10
2.3.5 Options ........................................................................................................................... 10
2.4 General features ............................................................................................. 11
2.5 Scope of supply .............................................................................................. 12
2.6 Installation requirements, Environmental operating conditions ........................ 13
2.7 Connecting 2010/2020 with an external computer ........................................... 14
2.8 Re-packing to prevent damage during transport .............................................. 14
3 Warranty .............................................................................................................15
3.1 Liability ............................................................................................................ 15
4 Installation .........................................................................................................16
4.1 Un-packing and System Set up ....................................................................... 16
4.2 Power On ........................................................................................................ 16
4.3 Software Installation ........................................................................................ 17
4.3.1 ADI and ADAP................................................................................................................ 17
4.4 Switching off.................................................................................................... 17
5 2020 On-Board Software ...................................................................................18
5.1 Controls and System Setup............................................................................. 18
5.1.1 Description of the key functions ..................................................................................... 18
5.1.2 Display ............................................................................................................................ 19
5.1.3 System Setup Functions ................................................................................................ 20
5.1.4 Setup Menu Options ...................................................................................................... 21
5.2 QUICK measurements without test programming and storage ........................ 22
5.2.1 F1 Single Point ............................................................................................................ 22
5.2.2 F3 Kinetic .................................................................................................................... 23
5.3 Test Definition - Programming of tests (evaluation modes) ............................. 25
5.3.1 Test Identification and Pre-Measurement Functions: .................................................... 25
5.3.2 Plate Layout Pattern ....................................................................................................... 26
5.3.3 Plate Layout ................................................................................................................... 28
5.3.4 Variables ........................................................................................................................ 30
5.3.5 Pre-Measurement Functions .......................................................................................... 31
5.3.6 Wavelength Selection .................................................................................................... 32
5.3.7 Kinetic Absorbency Measurement ................................................................................. 33
5.3.8 Relative Calculation ....................................................................................................... 34
5.3.9 Quantitative Evaluation .................................................................................................. 36
5.3.10 Qualitative Evaluation .................................................................................................... 40
5.3.11 Replicate Elimination ...................................................................................................... 42
5.3.12 Validation ........................................................................................................................ 43
5.3.13 Printing & Safeguarding the Test in the Test-Memory ................................................... 45
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Safety Information
All Warnings and Cautions in this document include an exclamation point, a
lightning bolt, or a light burst symbol framed within a triangle. Please pay special
attention to the specific safety information associated with these symbols.
WARNING
A WARNING calls attention to a condition or possible situation that
could cause injury to the operator.
CAUTION
A CAUTION calls attention to a condition or possible situation that
could damage or destroy the product or the operators work.
Electrical Safety
To prevent electrically related injuries and property damage, properly inspect all electrical
equipment priors to use and immediately report any electrical deficiencies. Contact an
Anthos service representative for any servicing of equipment requiring the removal of
covers or panels.
High Voltage:
This symbol indicates the potential of an electrical shock hazard
existing from a high voltage source and that all safety instructions
should be read and understood before proceeding with the installation,
maintenance, and servicing of all modules.
Voltages dangerous to human life are present in this device. Before removing any
covers disconnect the device from the power source.
Ensure that the power cord supplied with the unit is used.
The power cord may only be inserted in a socket outlet provided with a protective
ground (earth) contact. The protective action must not be negated by use of an
extension cord without a protective grounding contact.
Do not replace fuses without first removing the main power cord. Ensure that only fuses
with the required rated current and of the specified type are used for replacement. The
use of makeshift fuses and the short-circuiting of fuse-holders is prohibited.
When the apparatus is connected to the main power source, the opening of the covers
or removal of components is likely to expose live parts. The device shall be
disconnected from all voltage sources before it is opened for adjustment or repair
Any adjustment or repair of the opened apparatus under voltage should be avoided,
but, if necessary, it must be carried out by qualified service personnel who are aware of
the hazards involved.
Use the equipment only in the intended manner and as specified by the manufacturer,
otherwise the protection provided by the equipment may be impaired.
Observe all cautionary information printed on the original solutions containers prior to
their use.
Dispose of all waste solutions according to your facilitys waste disposal procedures.
Operate the 2010 and 2020 Absorbance Detectors in accordance with the instructions
outlined in this manual, and take all the necessary precautions when using
pathological, toxic, or radioactive materials.
Splashing of liquids may occur; therefore, take appropriate safety precautions, such as
using safety glasses and wearing protective clothing, when working with potentially
hazardous liquids.
Observe the appropriate cautionary procedures as defined by your safety officer when
using flammable solvents in or near a powered-up instrument.
Observe the appropriate cautionary procedures as defined by your safety officer when
using toxic, pathological, or radioactive materials.
Wash your hands thoroughly after handling test fluids. If equipment has been in contact
with hazardous substances, it must be disinfected prior to shipment in accordance with
the effective provisions.
Application:
Make sure that the plate is inserted correctly with well A1 on the upper left position.
Always consider that the reader is an optical instrument and is using the liquid surface
and the well bottom as optical surfaces.
Avoid foaming and make sure that no bubbles or particles in the well can disturb the
measurements. Extremely strong reactions may cause substrates to precipitate.
Visually inspect the bottom of plates before use for optical distortion and contamination.
Use bichromatic reading for eliminating unspecific signals caused by plate material or
contamination.
When using plate frames for column/row strips or single wells (breakable strips) check
that all strips and wells are pushed down completely and are level with the frame.
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Keep the bottom of the plate clean and dry. If liquid has contaminated the well bottoms,
wipe off with a soft cloth or paper suited for optical surfaces. Never touch the optical
surfaces with your fingers.
Do not insert plates into the reader, which are colder than the instrument and
environment. If this is required by the application, check the well bottoms for possible
condensation before reading.
Moving Parts
Do not touch the plate during movement of the plate transport (risk of injury).
Keep the 2010 and 2020 Absorbance Detectors work area clear to prevent obstruction
of the movement
.
1 Introduction
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2 Technical Information
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2.3.4 Interfacing
Parallel printer interface for IBM or EPSON compatible 9
and 24 pin printer and HP DeskJet 500 series (2020
only).
Serial interface for remote control of basic measurement
functions, instrument status and setup by external PC.
2.3.5 Options
A wide range of filters
PC-Software
Serial interface cable
Measurement Modes:
Evaluation possibilities:
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Qualitative evaluation:
Quantitative evaluation:
Combined evaluation:
Please Note:
Only use the power cable supplied with the instrument or a power
cable with protective earth connection carrying the CE-mark.
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Please Note:
The power indicator does not indicate that the instrument is turned
on or off or that it is connected with mains but only that the
instrument is ready for operation!
Connect the other end of the serial cable with the plug
marked "RS232" on the rear side of the instrument. Lock
the connector to the plug using the corresponding screws
on the connector.
Please Note:
Models 2010 and 2020 use different cables!
Only use the original cables supplied by anthos for 2010 (order
number S VZ 000 001) or 2020 (order number S VZ 000 002).
In emergency a standard 0-modem cable for PC-PC connection may
work with Model 2020 only, but not with Model 2010.
All other cables with connectors looking identical must not be used
to try to establish communication!
3 Warranty
A warranty period of 12 months shall be granted to the
original buyer of the anthos 2010/2020. This warranty shall
lose effect if:
anthos 2010/2020 is not used in the defined scope of
application,
anthos 2010/2020 has obviously been damaged by
external influences which are not in accordance with the
provisions for the nominal range of use,
anthos 2010/2020 has been modified or parts
exchanged by a person other than anthos personnel or
an authorized servicing agent,
the warranty seals on the housing of the instrument are
broken,
parts and subassemblies are implemented, which are
not original from Anthos,
anthos 2010/2020 serial number is no longer legible,
has been removed or altered,
anthos 2010/2020 has not been installed in accordance
with the instructions supplied,
anthos 2010/2020 has been damaged during return
transport due to wrong packing (e.g. not in original
packing material).
anthos 2010/2020 was damaged due to wrong
operation, not according to the descriptions in the
manual.
3.1 Liability
In its original condition the instrument meets all safety
regulations for a risk-less operation.
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4 Installation
4.2 Power On
Switch on main switch (rear left side). The anthos 2010
is immediately ready to stand by for remote control.
The anthos 2020 performs the complete initialization and
shows the main menu after approx. 30 seconds.
The power indicator is located on the front left side of the
unit and is marked with the following symbol.
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5.1.2 Display
Options:
Press <menu> and use the cursor keys to select menu
options
Function keys:
F1 - QUICK
Start a quick measurement without test programming
F2 RUN-TEST
Start a pre-programmed test
F4 INIT
Perform an initialization of the instrument
F5 SHUTDOWN
Prepares the instrument to be turned off
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Options:
Date and time (important for printed protocols)
Free Memory: shows the amount of free (available)
space on the hard drive or flash disk
Printer Type:
possible choices for printers are:
Matrix 9-pin
Matrix 24-pin
Inkjet Canon & Epson
Inkjet HP & others
Laser
IBM Proprinter
Postscript
HP Paint jet
HP Think jet
Floppy Disk Drive: to activate/deactivate press <ce>, an
activated Floppy Disk Drive is marked with [X]. To
activate functions regarding to data management using
disks , restart the machine with activated Floppy Disk
Drive.
Filter position and wavelength: To change, delete wrong
entry with CE and enter new value using the numerical
keyboard.
Function keys:
F1 Print Cfg
Print the configuration of the instrument
F2 Save
F3 am/pm
Switch between am and pm (only when Date Format is
set to 12 h mode, see below)
F5 Set Filters
After changing one or more filters always perform F5
Set Filters to make sure that the changed filter
configuration is stored and all filters will work with the
correct lamp energy settings!
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Function keys:
F1 Single Pt
Perform a single point, mono- or bichromatic absorbency
reading
F3 Kinetic
Perform a kinetic absorbency reading
Options:
Measurement filter: highlight the desired measurement
filter and confirm with <enter>
Reference filter: highlight the desired reference filter (as
above; if no reference filter is required, select ---)
Function keys:
F5 Start Meas.
Start the measurement with the selected filter(s).
5.2.2 F3 Kinetic
Options:
Measurement filter: highlight the desired measurement
filter and confirm with <enter>
Reference filter: highlight the desired reference filter (as
above; if no reference filter is required, select ---);
Usually no reference filter is used for kinetic data since a
reference reduction only shifts the relevant reaction
process up or down, which has no effect on the results
of the data reduction.
Interval: interval setting in seconds from the beginning of
one measurement cycle to the beginning of the next
cycle (minimum time = 45 seconds for single
wavelength, 90 seconds for dual wavelength).
Number of cycles: number of measurement cycles to be
performed (press <ce> and enter number using
keyboard).
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Data Reduction:
Possible choices are:
Delta OD
Average Slope
Maximum Slope
dOD Max. Slope
dt Max. Slope
dt Relative
dt Absolute
Please refer to chapter 5.2.2.1 for detailed information
on the different data reduction modes.
Smoothing Points:
Depending on the selected data reduction mode,
smoothing points can be defined: To reduce peaks in the
result, a number of values can be defined, of which an
average or slope (depending on the data reduction
mode) is calculated (e.g. if 3 smoothing points are
selected, an average or slope for the values 1, 2 and 3 is
calculated, then the average or slope for the values 2, 3
and 4 is calculated, etc.).
Function keys:
F5 - Start Meas.:
By pressing this function key the measurement will be
started with above defined parameters.
Delta t - Relative: Time in [sec] from the first
measurement to reaching a set increase/decrease in
[O D].
Delta t - Absolute: Elapsed time from one pre-selected
threshold value to another.
Function keys:
F5 Start Meas.
Start the measurement with the selected filter(s).
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Options:
Blank Validity
Validity of the blank value positions on the plate:
Plate: Mean value of all individual blanks is
automatically deducted from all other measurement
results
Row: Free configuration of the blanks on the entire
plate. The mean value of all blanks in each row is
deducted from each sample of this row.
Example: Setting blanks on plate position A01 and A08.
Mean value of A01 and A08 = (A01+A08)/2
this mean value is deducted from each sample of this row
(A02/A03/A04/A05/A06/A07/A09/A10/A11/A12).
Column: Free configuration of the blanks on the
entire plate. The mean value of all blanks in each
column is deducted from each sample of this
column.
Example: Setting blanks on A01, B01 and D01.
Mean value of A01, B01 and D01 = (A01+B01+D01)/3
this mean value is deducted from each sample of this column
(C01/E01/F01/G01/H01).
Fill Direction:
Filling/pipetting direction (horizontal/vertical).
Note: Groups of replicates are sorted according to their replicated
direction AND the filling direction.
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Options:
Well Type:
Select one of the possible well types (Standard, Control,
Blank,...). Press <F1 Layout> to jump to the plate
layout, position the cursor into the desired well and
press <enter> to define this well as the selected well
type. To select a different well type press <F1 Well
Type> and select with the cursor keys.
In the well type window the ID-number of the standard,
control, etc... can be increased by pressing <F2
Number +>.
Function keys:
F1 Layout/Well Type
By pressing this key the cursor can be set from the Well
Type window into the Layout window and vice versa. To
place the selected well type in the highlighted position of
the layout, press the enter key.
F2 Number +
Press this key to increase the ID-number of the selected
well type (control, standard, sample,...).
F3 Sample-Fill (Sample-Clr)
Press this key for automatic sample positioning. The
vacant positions are filled with samples in the prescribed
filling direction. After filling all positions, the function key
will change to Sample-Clr, which allows to clear all
samples set previously.
F4 Back
Return to the previous screen.
F5 - Next
Proceed to the next screen.
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5.3.4 Variables
For the evaluation of measurement results it is sometimes
necessary to use lot dependent factors or concentrations
(e.g. lot dependent titers of standards). For this reason up
to 24 variables per test can be defined and may be used for
further calculations.
Options:
Name:
Define a name for the desired variable by pressing <ce>
and enter the name with the cursor keys.
Value:
Enter the value for the selected variable.
Function keys:
F4 Back
Return to the previous screen.
F5 - Next
Proceed to the next screen.
Options:
Waiting Duration:
Enter the waiting time (waiting prior to measurement) in
seconds (1 999).
Function keys:
F4 Back
Return to the previous screen.
F5 - Next
Proceed to the next screen.
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Options:
Function keys:
F4 Back
Return to the previous screen.
F5 - Next
Proceed to the next screen.
Options:
Function keys:
F4 Back
Return to the previous screen.
F5 - Next
Proceed to the next screen.
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Options:
Result Name
Activate the text input with <ce> and enter a result
name.
Formula
Setting all plate positions in relation to a predefined
value.
This value is calculated according to an algebraic formula,
which may contain all the standards, controls and blanks
defined in the plate layout as well as the fundamental
operations of arithmetic and numerical constants and the
previously defined variables. Press <ce> to activate the
formula input.
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Function keys:
F4 Back
Return to the previous screen.
F5 - Next
Proceed to the next screen.
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Extrapolation:
The concentration of samples having a higher / lower
measurement result than the highest / lowest standard
can be determined mathematically. A continued
calculation of the curve can be performed up to 40%
outside the real curve limits.
This percentage corresponds to the concentration range
between the lowest and highest standard (=100%).
Extrapolated values are marked with an X on the
printout. If a curve loses its uniformity in the
extrapolation range (it reaches its maximum or
minimum), extrapolation is blocked.
Enter a value between 0 and 40 on the numerical
keyboard and confirm with <enter>.
2. Point to point:
Direct connection of all standard points.
Minimum number of defined standard points: 2
3. Cubic spline:
All standard points are connected by the best fitting
curve. Can only be used for non-linear and non-sigmoid
functions.
Minimum number of defined standard points: 3
Definition of the natural cubic spline as found in: G.
Engeln-Mllges, F. Reutter: Formelsammlung fr
numerische Mathematik mit Pascal-Programmen,
Bibliographical Institute, Mannheim, Vienna, Zurich;
Wissenschaftsverlag, 1985; p146ff.
4. 4 Parameter Fit:
This procedure can only be used to characterize sigmoid
curves. Minimum number of defined standard points: 4
The curve is calculated according to the following
formula:
(a d )
yi d
x
[1 ( i ) b ]
c
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Options:
Base
Select the base values (original measurement values or
results of the relative calculation by selecting the Result
Name or concentrations) for the qualitative evaluation. If
quantitative and qualitative evaluation is combined, the
measurement data can also be displayed in
concentrations and evaluated qualitatively as such by
using the standard curve.
Interpretation Mode
Set the type of threshold value subdivisions
1. Normal:
Up to four thresholds (cutoffs) can be programmed
individually and up to 5 groups selected.
2. Auto-ranging:
Upper and lower threshold are programmed. The
intermediate range can be divided into 2 to 10
equidistant groups.
3. %-ranging:
Upper and lower limits are programmed as cutoffs.
The sample measurement values are printed also
as percentages on the printout (the sample
measurement values are scaled in percent).
Number of Groups
Only appears for Interpretation Mode: normal. Enter
the number of groups to be subdivided (minimum=2,
maximum=5).
Number of Divisions
Only appears for Interpretation Mode: Auto-ranging.
Enter the number of equidistant tolerance value
subdivisions (minimum=2, maximum=10).
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Options:
Column Base
Select the base values (original measurement values or
results of the relative calculation by selecting the Result
Name or concentrations) for the calculation for
eliminating replicates.
Column Type
Definition of the standard / control / blank /sample (Spl)
which is to be evaluated with the following condition.
Press <ce>, select the corresponding position and
confirm with <enter>.
Column Min.
Enter the minimum number of remaining replicates (for
samples only 0 is possible).
Column Condition
Enter one ore more conditions, which must be fulfilled by
each replicate to avoid elimination. Press <ce> to
activate the input menu:
All special positions
defined in the plate layout (standards, controls,
blanks)
Arithmetic functions
(+, -, *, /. (), <, >, =, %)
Logical combinations
(NOT = true if condition is not fulfilled, AND = true if
all conditions are fulfilled, OR = true if one or more
conditions are fulfilled, XOR = true if only one of
the conditions is fulfilled)
Variables (please refer to chapter 5.3.4)
X (represents each individual replicate of the type
selected).
Move cursor to select a row and confirm with the
corresponding function key. Delete the last entry with <ce>.
Finally re-confirm the formula with <enter>.
Function keys:
F4 Back
Return to the previous screen.
F5 - Next
Proceed to the next screen.
5.3.12 Validation
If a programmed condition is not fulfilled, the entire test is
marked invalid.
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Options:
Column Base
Select the base for the calculation.
Column Condition
Enter one ore more conditions, which must be fulfilled by
the entire test to be valid. Press <ce> to activate the
input menu:
All special positions
defined in the plate layout (standards, controls,
blanks)
Arithmetic functions
(+, -, *, /. (), <, >, =, %)
Logical combinations
(NOT = true if condition is not fulfilled, AND = true if
all conditions are fulfilled, OR = true if one or more
conditions are fulfilled, XOR = true if only one of
the conditions is fulfilled)
Variables (please refer to chapter 5.3.4)
Move cursor to select a row and confirm with the
corresponding function key. Delete the last entry with <ce>.
Finally re-confirm the formula with <enter>.
Function keys:
F4 Back
Return to the previous screen.
F5 - Next
Proceed to the next screen.
5.3.13.1 Printing
Options:
Function keys:
F2 -->
Move the selected printing option from the Available
into the Chosen window.
F3 <--
Move the selected printing option from the Chosen into
the Available window.
F4 Back
Return to the previous screen.
F5 - Finish
Finish defining or editing the test and proceed to the
storage menu.
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Options:
Select the options with the cursor keys and confirm the
selection with <enter>.
Yes
Exit the define/edit mode and store the test in the test
memory of the instrument
No
Exit the define/edit mode without storing the test. All
definitions/changes will be discarded!
Cancel
Return to the define/edit mode (Finish screen).
Options:
Test Name
Press <ce> and choose the test to be started.
Plate-ID
An automatic plate ID is available in default
(YYMMDDxx, incremented from 1 to 99). To enter
another plate ID, press <ce> to delete the default
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Function keys:
Function keys:
F1 Print
Prints information concerning the test and measurement
time. Also, the results are printed according to the
settings in the test definition (please refer to chapter
5.3.13.1). Make sure a printer is connected and on-line,
and that the correct printer is selected in the system-
setup.
F2 Save As
Appears only if a plate had been re-evaluated. This
function enables to store the plate with a new name.
F5 Select Screen
Function keys:
F1 Well-Graph
Displays the reaction curve of the selected well.
F2 Plate-Graph
Displays the reaction curves of all well of the plate.
F3 Prev RawDat
Displays the raw data of the previous measurement.
F4 Next RawDat
Displays the raw data of the next measurement.
F5 Select Screen
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Function keys:
F1 Re-Calc
To re-calculate the plate in case that the rejection state
of one ore more wells had been changed.
F5 Select Screen
Function keys:
F5 Select Screen
Function keys:
F5 Select Screen
Function keys:
F1 Re-Calc
To re-calculate the plate in case that curve parameters
had been changed.
F3 Graphics
Displays the standard curve according to the defined
parameters.
F5 Select Screen
Function keys:
F5 Select Screen
5.6.9 CV % Screen
This screen shows the percentage correlation of variation of
replicate standards, controls and samples. The value is
shown on the upper left plate position of replicates.
Function keys:
F5 Select Screen
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evaluated. Select the test with which the plate shall be re-
evaluated.
Note: Please make sure to have printouts of the result available prior to
deleting it!
Copy: The copy option copies the data from the anthos
2020 to the disk; the data is now available on both,
the instrument and the disk.
Shift: The shift option moves the data from the storage of
the anthos 2020 to the disk; the data is now available
only on the disk.
Function keys:
F1 Del Plate (Del Test / Del Result)
Deletes the highlighted data (plate, test or result)
F2 Del all Plt (Del all Tst / Del all Res)
Deletes all data in the selected (highlighted) list (plates,
tests or results)
F3 Delete All
Deletes all data (all plates, tests and results) on the
floppy disk.
F5 - Refresh
Re-reads the data stored on the inserted floppy disk.
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6 Maintenance
The following materials are recommended for cleaning and disinfection of the
reader:
Non-lint tissues.
6.2.1 Cleaning
Clean the reader regularly and immediately after spillage.
This has to be done with due care and attention. Always
observe laboratory safety rules and regulations. Do not use
force when cleaning the reader.
Wipe off spills immediately with soft tissue.
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6.2.2 Disinfection
Before the reader is returned to the distributor it must be
disinfected and a disinfection certificate must be completed.
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7 Appendix:
Transmission is not related to layer thickness (path-length of the light) and sample
concentration in a linear way:
If one layer of a certain material allows 10% transmission, doubling either path-
length or concentration would bring down the transmission to 1% and doubling both
would bring it down to only 0.01% (logarithmic relation)!
Absorbency is related in a linear way to layer thickness and sample concentration
according to the "Bouger-Lambert-Beer" law:
E = OD = c * d * e
(c = the concentration of the sample to be measured
d = the layer thickness of the sample to be measured
e = the molar extinction coefficient)
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7.2.1.11 Validation:
The validation formulas describe true conditions on basis of optical density values.
A bold warning will be stated on the printout, if one of the validation criterias is not
fulfilled.
Copy the original test procedure to a new file by changing the name and editing the test
(a copy will be stored under the new name).
The layout does not need to contain a real standard anymore. Fill the plate just with
samples
Standard Points S1 S2 S3 S4 S5
Concentration 0.1 1 10 100 1000
Values
Response Formulas S1 S2 S3 S4 S5
A new test is programmed: The measured ODs of the standards are entered to the
variables:
Standard Points S1 S2 S3 S4 S5
Response Values 0.3 0.6 0.8 0.9 1.0
[OD]
Put to Variables V1 V2 V3 V4 V5
Only standard S3 is used on the plate-layout of the new test. It is named C1 (control)
and its measured value is used to calculate the correction factor for all other
standards:
Standard Points S1 S2 S3 S4 S5
Concentration 0.1 1 10 100 1000
Values
Response V1*(C1/V3 V2*(C1/V3 V3*(C1/V3 V4*(C1/V3 V5*(C1/V3
Formulas ) ) ) ) )
If the actual measurement value of the standard 3 (C1) deviates from the original
measurement value (V3), all standards are re-adjusted by the correction factor
C1/V3 corrected:
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anthos 2010/2020 Operating Manual
Most probably, the total kinetic measurement time can be set shorter than the
normal substrate incubation time. TMB measured in the blue phase at 620nm has
about one third of the optical density than the same concentration measured after
stopping in the yellow phase. Samples and standards, which would reach
"overflow" in an endpoint reading, will remain within the good measurement range
during kinetic readings. Therefore the dynamic range and accuracy of ELISAs may
be significantly improved with kinetic readings.
A main advantage of kinetic readings is the elimination of the critical timing between
adding of substrate and stopping solution for all wells.