PlateReader ual AF2200 EN) manual Register your instrument! www.eppendorf.com/myeppendorf Eppendorf® PlateReader AF2200 Software manual Version 1.0.
Copyright© 2013 Eppendorf AG, Hamburg. No part of this publication may be reproduced without the prior permission of the copyright owner. Trademarks Eppendorf® and the Eppendorf Logo are registered trademarks of Eppendorf AG, Hamburg, Germany. Microsoft®, Windows® and Excel® are registered trademarks of Microsoft Corporation, Redmond, WA, USA. Pentium® is a registered trademark of Intel Corporation, Santa Clara, CA, USA.
Table of contents Eppendorf® PlateReader AF2200 English (EN) Table of contents 1 Operating instructions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1.1 Using this manual . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1.2 Danger symbols and danger levels . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1.2.
Table of contents Eppendorf® PlateReader AF2200 English (EN) 6 Predefined methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6.1 Method selection. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6.2 Method description . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6.2.
Operating instructions Eppendorf® PlateReader AF2200 English (EN) 1 1.1 Operating instructions Using this manual Read this operating manual completely before using the device for the first time. Please also note the operating instructions for the accessories, if applicable. This operating manual is part of the product. Thus, it must always be easily accessible. Enclose this operating manual when transferring the device to third parties. If this manual is lost, please request another one.
Operating instructions Eppendorf® PlateReader AF2200 English (EN) 1.3 Symbols used Symbol Meaning Handling 1. 2.
Product description Eppendorf® PlateReader AF2200 English (EN) 2 2.1 Product description Area of Application The Software is an easy-to-use and flexible tool, which gives the user complete control over the Eppendorf PlateReader AF2200. The Software presents the raw data for further use in Excel, offering excellent features for research purposes. 2.
Product description Eppendorf® PlateReader AF2200 English (EN) Component Minimum Recommended Communication 1x USB 2.0 2x USB 2.0 1x RS232 (serial) Drive, graphic card 1xCD-ROM drive Windows Vista: DirectX 9 graphic and 32 MB graphics memory (for Home Basic version) DirectX 9 graphic and 128 MB graphics memory with WDDM support for all other versions. Windows 7: DirectX 9 graphic with WDDM driver 1.0 or higher. .NET Microsoft .NET Framework 2.
Installation Eppendorf® PlateReader AF2200 English (EN) 3 3.1 Installation Install software You must have administrator rights to install the software. Install the software before connecting the PlateReader AF2200 to the computer. 1. Insert the CD with the software into the CD-ROM drive. 2. Start the installation by executing the Setup file. 3. Press the OK button to continue the installation. 4. Press the Next button to continue the installation.
Installation Eppendorf® PlateReader AF2200 English (EN) 5. Define the user. 6. Select language. 7. Define the installation directory.
Installation Eppendorf® PlateReader AF2200 English (EN) 8. Press the Install button to confirm the settings and start the installation. 9. Press the Finish button to complete the installation. The PlateReader AF2200 can now be connected to the computer.
Installation Eppendorf® PlateReader AF2200 English (EN) 3.2 Starting PlateReader AF 2200 The Software can be used either with a connected instrument or in simulation mode. 3.2.1 Connected Instrument Install the software before connecting the instrument to the computer. 1. Connect the instrument to your computer and switch the instrument on. 2. Start the program by selecting Programs > Eppendorf > PlateReader AF2200 from the Windows Start menu. 3. The following dialog box appears: 4.
Overview Eppendorf® PlateReader AF2200 English (EN) 4 4.1 Overview Main Window The main window of the software is used to set up workflows. Each workflow is easily created by dragging and dropping the process steps into a sequence according to the application. The application workflow is then visible to the user in the workflow pane and can be saved for future use. Each process step, that is each program element, can be copied and pasted and moved to the desired position in the workflow.
Overview Eppendorf® PlateReader AF2200 English (EN) 4.2 Explorer bar The Explorer bar is divided into different sections. Each section contains program elements used to create an individual workflow. 1. Create a workflow either by double-clicking the selected program element or by dragging and dropping it into the workflow pane.
Overview Eppendorf® PlateReader AF2200 English (EN) 4. Under Details it is possible to apply a filter so that only certain plate definition files are shown. Pattern The Pattern program element is displayed according to the selected plate format (number of wells). 1. To measure individual wells, click the desired well or to measure a range of wells drag a frame around the desired range. Independent Pattern 1.
Overview Eppendorf® PlateReader AF2200 English (EN) Well Use the Well program element to perform measurements well by well. Without this program element, all measurement steps are done plate-wise. 4.2.2 Mode Absorbance The Absorbance program element is used to perform absorbance measurements. 1. Enter or select the respective parameters: Wavelength Specify a measurement wavelength. The Reference wavelength may be selected to correct for flash variations.
Overview Eppendorf® PlateReader AF2200 English (EN) Fluorescence Intensity The Fluorescence Intensity program element contains fields for the selection of excitation and emission wavelength, top or bottom reading mode, integration and lag time, flash number and gain settings. A checkbox for multiple reads per well gives access to additional function. The following are the Fluorescence Intensity parameters: Wavelength and Bandwidth Specify an Excitation and an Emission wavelength.
Overview Eppendorf® PlateReader AF2200 English (EN) Gain The gain is an amplification factor for the photomultiplier tube (PMT) and may be set by selecting one of the following modes: Manual gain: user-defined gain value (valid range: 1-255) Optimal gain: calculated automatically by the instrument according to the highest signal within the selected well range in order to avoid OVER. Optimal gain determination is performed in a pre-measurement.
Overview Eppendorf® PlateReader AF2200 English (EN) Multiple Reads per Well The software allows the user to define multiple reads per well (MRW) in Absorbance, Fluorescence top and Fluorescence bottom mode. The MRW feature is not available for well wise measurements. The Reference wavelength on the absorbance program element is not selectable in combination with multiple reads per well.
Overview Eppendorf® PlateReader AF2200 English (EN) Shaking 1. Select the Shaking program element if the plate is to be shaken, either before the measurement or between kinetic cycles. 2. Enter the respective parameters (see Parameter list). 3. Clicking the link Wait a couple of seconds inserts a new program element (see Miscellaneous on p. 21). Tab. 4-1: Parameter list Duration Enter the duration of the shaking process. Mode Select between the options Linear and Orbital.
Overview Eppendorf® PlateReader AF2200 English (EN) Well-wise kinetic measurements All cycles of the kinetic measurement are first performed in one well before continuing to the next well. Well-wise kinetic measurements may be composed of a maximum of four measurement stripes of the same type, e.g., four absorbance stripes. Kinetic Condition Use the Kinetic Condition program element to define which actions should be executed at a certain cycle.
Overview Eppendorf® PlateReader AF2200 English (EN) User Intervention The User Intervention program element informs the operator of the instrument to execute a definite action during the workflow at a certain time. If for example the Move Plate program element is used to move the plate out to perform a certain action, then the entered text should inform the operator to perform these actions. A dialog box shows the message and the measurement process stops until OK is clicked.
Overview Eppendorf® PlateReader AF2200 English (EN) Incubation Incubation is always done at the heating position to ensure inside the instrument proper temperature distribution. Incubation can consist of shaking and waiting steps (up to 2 shaking steps and up to 2 waiting steps are allowed in any combination). The Remaining Wait step waits until the overall incubation time is over (including shaking and waiting times).
Overview Eppendorf® PlateReader AF2200 English (EN) 4.3 Workflow Pane The main window in the software is the Workflow pane, where the measurement script is visible and where parameters are defined and edited. There are two ways to insert a program element from the Explorer bar into the Workflow pane. 1. Select a program element from the Explorer bar; by double-clicking it, it is inserted into the Workflow pane directly after the previous program element. 2.
Overview Eppendorf® PlateReader AF2200 English (EN) 4.3.1 Hierarchy of Elements The hierarchy of elements in the Workflow pane is as follows: • Plate • Pattern • Well Any desired measurement step can be inserted directly after a plate, a pattern or a well element. Arrows placed on the left side of the program elements header to release or indent it.
Overview Eppendorf® PlateReader AF2200 English (EN) 4.5 Menu Bar 4.5.1 File Menu New This command opens a new measurement workflow. If an empty document is to be opened, you will be asked to save the current workflow. • Click Yes to save the current workflow. • Click No to create a new workflow without saving the previous one. • Click Cancel to leave the dialog box. Open This command opens an existing PlateReader AF2200 Software workflow (*.mdfx) from the selected folder.
Overview Eppendorf® PlateReader AF2200 English (EN) Properties Possibilty to save information for a script or a method. Save This command saves the current script. Save As… This command saves the current workflow under a different name. List of most recently used script files A list of the most recently saved workflow files is displayed. Define how many files are to be included in this list in the Settings menu > User settings. Exit This command exits and closes the program.
Overview Eppendorf® PlateReader AF2200 English (EN) 4.5.3 View Menu Status bar This command shows or hides the status bar (located at the bottom of the window). Collapse All This command collapses all program elements in the workflow pane to view only one line of text. Expand All This command expands all program elements in the workflow pane to extended view and shows all visible parameters. 4.5.
Overview Eppendorf® PlateReader AF2200 English (EN) Heating This command is used to set the target temperature of the instrument manually. • Select or enter the Target temperature and click Set and On to start instrument heating. • Click the Read button to display the current temperature inside the instrument or click the Auto check box to have it read automatically. • Click Off to stop heating.
Overview Eppendorf® PlateReader AF2200 English (EN) Filter Definitions Purchase Date This option enables the user to enter the purchase or installation date of the filter. Flash Counter The flash counter monitors the number of flashes through a filter. The flash counter number provides the user only with additional information about the filter in use. • For a new filter, set the counter to 0. • For a previously used filter, enter the last collected flash number if the number is available.
Overview Eppendorf® PlateReader AF2200 English (EN) Plate Definition This command allows you to choose a plate file from the drop-down list of available plates. The plate definition files contain all relevant parameters of a specific plate type, e.g. coordinates of measurement points, number of columns, number of rows, well form, well diameter, plate height, plate height with cover…). A graphic element at the bottom of the dialog visualizes the parameter which is currently defined.
Overview Eppendorf® PlateReader AF2200 English (EN) User Settings User Settings - Tab Start Up Behavior at start up can be set. 1. Select a default plate. 2. Determine if the workflow pane should start with an empty workflow, plate only, or plate and part of plate. 3. Select whether the last used instrument should be reconnected 4. Select whether the Open Template dialog at startup should be skipped. User Settings - Tab General General options can be set. 1.
Overview Eppendorf® PlateReader AF2200 English (EN) User Settings - Tab Language 1. Select the language of the PlateReader AF2200 Software 2. Click OK to save your settings or click Cancel to leave the dialog box without saving any changes Data Presentation This command offers the following tabs to determine the output settings of the measured results in Excel.
Overview Eppendorf® PlateReader AF2200 English (EN) View Mode: Select between Matrix and List. • If Matrix is selected, the data alignment corresponds to a microplate; times per well cannot be displayed. Not relevant for kinetic result presentation. • If List is selected, choose between: Align, Rotation, Display Times. Show: Select between All and Measured. • If All is selected, the whole plate geometry, including all possible rows and columns, is displayed.
Overview Eppendorf® PlateReader AF2200 English (EN) Data Presentation - Eppendorf μPlate G 0.5 Show Raw Data: Select the Show Raw Data box to display the raw measurement values of a Nucleic Acid Quantification measurement. Exception History The Exception History dialog box shows a list of exceptions (instrument errors, software failures) with date and time. Every time an exception occurs and an error box is displayed, all relevant information is collected and saved in a zip-file.
Overview Eppendorf® PlateReader AF2200 English (EN) 4.6 Plate List Order No.
Overview Eppendorf® PlateReader AF2200 English (EN) Order No.
Overview Eppendorf® PlateReader AF2200 English (EN) Order No.
Overview Eppendorf® PlateReader AF2200 English (EN) Order No.
Overview Eppendorf® PlateReader AF2200 English (EN) Order No. Manufacturer Number of Wells Bottom Shape Color Additional Information 0030 621.905 Eppendorf 384 V Black - 0030 601.475 Eppendorf 96 Flat White - 0030 601.572 Eppendorf 96 U White - 0030 601.670 Eppendorf 96 V White - 0030 621.670 Eppendorf 384 V White - 0030 730.020 Eppendorf 96 F Clear VIS 0030 741.048 Eppendorf 96 F Clear UV-VIS 0030 730.119 Eppendorf 96 F Clear Cell Culture 0030 741.
Defining measurements Eppendorf® PlateReader AF2200 English (EN) 5 Defining measurements The following chapter describes some examples to illustrate the definition of different measurements. 5.1 Plate Size – Pattern 1. Use the Plate program element in the workflow pane to choose a plate format. 2. Select the desired plate format from the Plate definition drop-down list (e.g. Eppendorf 96 Flat Clear UV-VIS) 3. To measure a particular well or a range of wells on the plate click the link Pattern. 4.
Defining measurements Eppendorf® PlateReader AF2200 English (EN) 5.2 Defining Endpoint Measurements The following example describes an Absorbance Endpoint Measurement in all wells of a 96 well plate. Select a 96 well plate (e.g. Eppendorf 96 Flat Clear UV-VIS) from the Plate definition drop-down list. If the Pattern program element is not visible, click the link Pattern. It is recommended to use the Pattern program element in every workflow, even if all wells are measured.
Defining measurements Eppendorf® PlateReader AF2200 English (EN) If the plate shall be moved out of the instrument after measurement, insert a Move Plate program element and select the Out radio button. If a Move Plate program element is not defined after the measurement, the plate will stay inside the instrument until Move Plate Out is clicked. After finishing the definition as described above start the measurement by clicking the Start button on the right side beneath the Info bar.
Defining measurements Eppendorf® PlateReader AF2200 English (EN) 5.3 Defining Multimode Measurements Multimode measurements are measurements with multiple consecutive reading modes, e.g. with multiple absorbance and fluorescence measurements or with mixed measurements. The following example describes the definition of a Multimode measurement in a 96 well plate. Measurement 1 – Absorbance 462 nm in all wells 1. Select a 96 well plate (e.g.
Defining measurements Eppendorf® PlateReader AF2200 English (EN) Measurement 2 – Fluorescence 485/535 nm in all wells 1.
Defining measurements Eppendorf® PlateReader AF2200 English (EN) Measurement 3 – Fluorescence 360/465 nm in all wells 1. Insert a second Fluorescence Intensity program element from the Explorer bar and define as follows: • Wavelength/Excitation: 360 nm • Wavelength/Emission: 465 nm • Read/Number of reads: 25 • Gain: Optimal 2. After finishing the definition as described above start the measurement by clicking the Start button on the right side beneath the Info bar.
Defining measurements Eppendorf® PlateReader AF2200 English (EN) 5.4 Defining Kinetic Measurements The following example describes a kinetic measurement of a 96 well plate. Select the 96 well plate (e.g. Eppendorf 96 Flat Clear UV-VIS) from the Plate definition drop-down list, and select all wells in the Pattern program element.
Defining measurements Eppendorf® PlateReader AF2200 English (EN) After finishing the definition as described above start the measurement by clicking the Start button on the right side beneath the Info bar. When clicking the Start button, Excel opens automatically and the raw data are displayed in a worksheet.
Defining measurements Eppendorf® PlateReader AF2200 English (EN) 5.5 Indenting and Releasing Program Elements The decision to indent or release a program element will modify the workflow of the instrument during measurements. The actions of all program elements with the same indentation are performed sequentially. The only dependence between these program elements is that the next action starts directly after the previous action is finished.
Defining measurements Eppendorf® PlateReader AF2200 English (EN) The Workflow pane appears as shown in the screenshot. The above definition results in the following workflow. The Absorbance of all wells of a 96 well plate is first measured at 260 nm and then at 280 nm. Both Absorbance measurements are performed in 5 kinetic cycles. Indenting the second Absorbance program elements on a level with Kinetic Cycle item changes the workflow.
Defining measurements Eppendorf® PlateReader AF2200 English (EN) In this workflow, an Absorbance Kinetic measurement with 5 cycles is done first at 260 nm; finished this loop, Absorbance Endpoint measurement at 280 nm is performed. Ways to Indent or Release Script Elements Select a program element from the Workflow pane. Click Edit and Indent Stripe/Release Stripe Use the arrow buttons in the header of the program element to release or indent the selected element.
Defining measurements Eppendorf® PlateReader AF2200 English (EN)
Predefined methods Eppendorf® PlateReader AF2200 English (EN) 6 6.1 Predefined methods Method selection Frequently-used methods in molecular and cell biology are provided as predefined methods. To load a method, select the required method from the upper left "Methods" section of the Explorer bar either by double-clicking the method or by dragging and dropping the method into the Workflow pane. Abb. 6-1: Overview of predefined methods Fig.
Predefined methods Eppendorf® PlateReader AF2200 English (EN) 6.2 Method description 6.2.1 Nucleic acid quantification (UV 260 nm microvolume) This method is restricted to the Eppendorf μPlate G0.5. For microvolume measurements with the μPlate G0.5 please refer to the dedicated manual. 6.2.2 Nucleic acid quantification (UV 260 nm with factor) Introduction The most common technique for measuring nucleic acid concentration is based on measuring the absorbance at 260 nm.
Predefined methods Eppendorf® PlateReader AF2200 English (EN) Default measurement parameters • Measurement wavelength: 260 (5) nm • Ratio wavelength (for purity check with 260/280 ratio): 280 (5) nm, optional • Background wavelength (for correction of turbidity): 340 (10) nm, optional • Number of flashes: 25 • Settle time: 0 ms • Plate definition: Eppendorf Microplate UV-VIS, 96/F The method is restricted to Eppendorf Microplate UV-VIS, 96/F. The pathlengths are determined for aquaeus buffer.
Predefined methods Eppendorf® PlateReader AF2200 English (EN) Plate layout Abb. 6-2: Method strip: Nucleic acid quantification (UV 260 nm with factor) Fig. 6-2: Method strip: Nucleic acid quantification (UV 260 nm with factor) The selected number of blanks and samples are shown in the plate view on the right side of the method strip. 6.2.
Predefined methods Eppendorf® PlateReader AF2200 English (EN) User defined method parameters • The measurement is performed at 260 nm with default measurement parameters. • Define the number of blank replicates (possible number: 1 – 8). • Define the number of standards (possible number: 2 – 12). • Define number of standard replicates (possible number: 1 – 8). • Define the concentrations of each standard (Press triangle button to open the menu). • Define the number of samples (max.
Predefined methods Eppendorf® PlateReader AF2200 English (EN) Plate layout Abb. 6-3: Method strip: Nucleic acid quantification (UV 260 nm with standards) Fig. 6-3: Method strip: Nucleic acid quantification (UV 260 nm with standards) The selected number of blanks, standards and samples are shown in the plate view on the right side of the method strip.
Predefined methods Eppendorf® PlateReader AF2200 English (EN) 6.2.4 Nucleic acid quantification (Fluorescence 485/535 nm with standards) Introduction PicoGreen is a highly sensitive fluorescent nucleic acid stain for the quantitation of double-stranded (ds)DNA in solution. The measured fluorescence intensity signal is directly proportional to the amount of dsDNA present in the sample. RiboGreen is one of the most sensitive stains for the detection and quantitation of RNA in solution.
Predefined methods Eppendorf® PlateReader AF2200 English (EN) Plate layout Abb. 6-4: Method strip: Nucleic acid quantification (Fluorescence 485/535 nm with standards) Fig. 6-4: Method strip: Nucleic acid quantification (Fluorescence 485/535 nm with standards) The selected number of blanks, standards and samples are shown in the plate view on the right side of the method strip.
Predefined methods Eppendorf® PlateReader AF2200 English (EN) 6.2.5 Protein quantification BCA, Bradford, Lowry BCA 562 (600) nm with standards, Bradford 595 (600) nm with standards, Lowry 750 (600) nm with standards Introduction The BCA Protein Assay uses bicinchoninic acid (BCA) for colorimetric quantification of total protein in a sample. The method is based on the reduction of Cu2+ to Cu1+ by protein in an alkaline medium.
Predefined methods Eppendorf® PlateReader AF2200 English (EN) User defined method parameters • Using the predefined UV/Vis filter slide the measurement is performed at 600 nm with default measurement parameters. • Define the number of blank replicates (possible number: 1 – 8). • Define the number of standards (possible number: 2 – 12). • Define number of standard replicates (possible number: 1 – 8). • Define the concentrations of each standard (Press triangle button to open the menu).
Predefined methods Eppendorf® PlateReader AF2200 English (EN) 6.2.6 Protein quantification (NanoOrange 485/595 nm with standards) Introduction NanoOrange (Molecular Probes) is a highly sensitive fluorescent stain for protein quantification in solution. The reagent is virtually non-fluorescent in aqueous solution, but when bound to proteins undergoes a strong fluorescence enhancement with a broad excitation peak centered at app. 470 nm and a broad emission peak centered at app. 570 nm.
Predefined methods Eppendorf® PlateReader AF2200 English (EN) Plate layout Abb. 6-6: Method strip: Protein quantification (NanoOrange 485/595 nm with standards) Fig. 6-6: Method strip: Protein quantification (NanoOrange 485/595 nm with standards) The selected number of blanks, standards and samples are shown in the plate view on the right side of the method strip.
Predefined methods Eppendorf® PlateReader AF2200 English (EN) 6.2.7 Cell viability (CellTiter-Blue Assay) Introduction The CellTiter-Blue Cell Viability Assay provides a homogeneous, fluorometric method for monitoring cell viability. The assay is based on the ability of living cells to convert a redox dye (resazurin) into a fluorescent end product (resorufin). Nonviable cells rapidly lose metabolic capacity and thus do not generate a fluorescent signal.
Predefined methods Eppendorf® PlateReader AF2200 English (EN) Plate layout Abb. 6-7: Method strip: Cell viability (CellTiter-Blue Assay) Fig. 6-7: Method strip: Cell viability (CellTiter-Blue Assay) The selected number of blanks, controls and samples are shown in the plate view on the right side of the method strip.
Predefined methods Eppendorf® PlateReader AF2200 English (EN) 6.2.8 Cell viability (Apo-ONE Caspase-3/7 Assay) Introduction The Apo-ONE Homogeneous Caspase-3/7 Assay provides a homogenous fluorometric method for monitoring apoptosis in mammalian cells. The assay is based on the measurement of the activities of the proteases caspase-3 and -7 in the process of apoptosis. A pro-fluorescent substrate becomes fluorescent after cleavage by the activated caspases.
Predefined methods Eppendorf® PlateReader AF2200 English (EN) Plate layout Abb. 6-8: Method strip: Cell viability (Apo-ONE Caspase-3/7 Assay) Fig. 6-8: Method strip: Cell viability (Apo-ONE Caspase-3/7 Assay) The selected number of blanks, controls and samples are shown in the plate view on the right side of the method strip.
Predefined methods Eppendorf® PlateReader AF2200 English (EN) 6.3 Method (measurement) parameters The methods were created with default measurement parameter sets which are listed in the method descriptions. In some cases it may be necessary to change parameter settings in order to reach optimal results. 6.4 Method results As the measurement is performed, an Excel Workbook opens automatically. It consists of a raw data sheet, a blanking sheet (factor methods only) and a results sheet.
Predefined methods Eppendorf® PlateReader AF2200 English (EN) 6.4.2 Contents of the blanking sheet (factor methods only) Blanking results and all other intermediate results. Abb. 6-10: Blanking sheet Fig.
Predefined methods Eppendorf® PlateReader AF2200 English (EN) 6.4.3 Content of the results sheet The structure of the results sheet depends on the method. Default contents are: • method parameters • plate layout with sample IDs • raw data, blanked raw data and sample results • method specific calculations • standard methods contain standard results incl. standard curve graph • cell viability methods contain a bar chart which shows a result overview The sample IDs in the plate layout are alterable.
Predefined methods Eppendorf® PlateReader AF2200 English (EN)
Troubleshooting Eppendorf® PlateReader AF2200 English (EN) 7 7.1 Troubleshooting General Troubleshooting If you don’t meet your expected measurement results please check the following: • The volumes of all measured wells are homogeneous. Uniformity of pathlength [d] is essential for reliable measurements. Identical volumes of different fluid types and buffers may have different pathlengths. d • The homogeneity of your samples is given: The bottom of the wells must be entirely covered with fluid.
Troubleshooting Eppendorf® PlateReader AF2200 English (EN) 7.3 Troubleshooting Fluorescence mode Warning / error Consequences Remedy Sample data out of min-max-range of standard values. Corresponding sample results are Dilute sample or modify standard marked red in the results table. concentrations and remeasure. Correlation coefficient of standard Sample results are not calculated. Check input of standard curve < user input. concentrations and remeasure.
Evaluation procedure Eppendorf® PlateReader AF2200 English (EN) 8 8.1 Evaluation procedure Nucleic acid quantification UV 260 nm with factor Sample results [Unit] = absorbance * factor sample type * correction factor pathlength 8.
Evaluation procedure Eppendorf® PlateReader AF2200 English (EN) Average raw data Average value of control replicates / blank replicates / sample replicates in RFU SD raw data Standard deviation of control replicates / blank replicates / sample replicates in RFU SD [%] raw data Standard deviation of control replicates / blank replicates / sample replicates in % Average blanked Average value blanked = Average value of control replicates / sample replicates – average value of blank replicates in RFU SD bl
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