PlateReader nual gEN) manualAF2200 Eppendorf® PlateReader AF2200 Operating manual
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. Greiner® is a registered trademark of Greiner Bio-One GmbH, Frickenhausen, Germany.
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) 5.3 Installing user-specific filters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5.3.1 Removing filters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5.3.2 Inserting user-specific filters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5.3.3 Defining filters. . . . . . . . . . . . . .
Table of contents Eppendorf® PlateReader AF2200 English (EN) 9 Technical data . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9.1 Power supply. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9.2 Weight/dimensions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9.
Table of contents Eppendorf® PlateReader AF2200 English (EN)
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 Depiction Meaning 1. 2.
Operating instructions Eppendorf® PlateReader AF2200 English (EN) 1.
Operating instructions Eppendorf® PlateReader AF2200 English (EN)
Product description Eppendorf® PlateReader AF2200 English (EN) 2 2.1 Product description Main illustration Abb. 2-1: Front and rear view 7 1 2 Fig. 2-1: 3 4 5 6 Front and rear view 1 Filter slide flap 5 USB port, USB connector Typ B 2 Button for moving the plate compartment in and out 6 Warranty seal 7 Name plate 3 Plate compartment flap 4 Mains connection socket with power switch 0 = switched off, I = switched on. The device may only be opened by the manufacturer's service technicians.
Product description Eppendorf® PlateReader AF2200 English (EN) 2.3 Features The PlateReader AF2200 is well-suited for the quantification of biomolecules, such as nucleic acids and proteins, and for measuring fluorescence-based assays in plate format. A wide variety of methods can be executed in the UV/Vis range, and various fluorescence dyes can be detected. The PlateReader AF2200 is designed for use in molecular biology, biochemistry and cell biology research laboratories.
Product description Eppendorf® PlateReader AF2200 English (EN) 2.
Product description Eppendorf® PlateReader AF2200 English (EN) Component Minimum Recommended 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.
Product description Eppendorf® PlateReader AF2200 English (EN) 2.5 Fluorescence detection system Abb. 2-2: Upper fluorescence detection system (fluorescence top reading) PMT reference diode 3 emission filter condensor beam splitter flash lamp 1 excitation filter 2 MTP 2 Fig.
Product description Eppendorf® PlateReader AF2200 English (EN) Abb. 2-3: Lower fluorescence detection system (fluorescence bottom reading) PMT reference diode 3 emission filter condensor flash lamp 1 2 excitation filter mirror Fig. 2-3: bottom emission fiber mirror MTP 16 bottom excitation fiber 2 Lower fluorescence detection system (fluorescence bottom reading) 1 Light source 3 Fluorescence detector 2 Fluorescence optics 2.5.
Product description Eppendorf® PlateReader AF2200 English (EN) Flash lamp The device uses a high-performance xenon gas-discharge lamp (flash lamp). The flash jumps over a small gap between two electrodes. The lamp contains high-pressure xenon. The flash disappears within several microseconds. The flash frequency is 40 Hz. The device uses the flash lamp for fluorescence and absorbance measurements.
Product description Eppendorf® PlateReader AF2200 English (EN) The lens system The lens system bundles the maximum amount of fluorescent light from a well and focuses it onto the detection system through the exit aperture. The lenses are made of quartz glass. This material is extremely UV-transparent and virtually non-autofluorescent. 2.5.3 Lower fluorescence optics Flash light enters the optical system focused through an aperture. The opening functions as a color-specific light source.
Product description Eppendorf® PlateReader AF2200 English (EN) 2.6 Absorbance measurement system A light path similar to the one used for fluorescence excitation is implemented for absorbance measurements. The absorbance measurement module is located below the plate compartment. It measures the light that is transmitted through the sample. A reference measurement, during which the plate compartment is positioned outside of the light beam, is carried out before the microplate is measured. Abb.
Product description Eppendorf® PlateReader AF2200 English (EN) 2.6.1 Light source system The absorbance light source system is similar to the upper fluorescence system (see p. 16). Absorbance filter For absorbance applications, optical band-pass filters are required to enable suitable wavelengths to pass through and to filter the rest of the flash lamp spectrum. The filters are mounted in removable slides.
Safety Eppendorf® PlateReader AF2200 English (EN) 3 3.1 Safety Intended use The PlateReader AF2200 is designed for use in molecular biology, biochemistry and cell biology research laboratories. The device was developed for use as a universal laboratory device for processing biological and non-biological samples. The PlateReader AF2200 is intended for indoor use only. All country-specific safety requirements for operating electrical equipment in the laboratory must be observed.
Safety Eppendorf® PlateReader AF2200 English (EN) WARNING! Damage to health from toxic, radioactive or aggressive chemicals as well as infectious liquids and pathogenic germs. Observe the national regulations for handling these substances, the biological security level of your laboratory, the material safety data sheets and the manufacturer's application notes. Wear personal protective equipment. CAUTION! Poor safety due to incorrect accessories and spare parts.
Safety Eppendorf® PlateReader AF2200 English (EN) NOTICE! Function impairment due to mechanical damage. After mechanical damage to the device, ensure that the measuring and evaluation functions of the device are operating correctly by completing an inspection. NOTICE! Damage from overheating. Do not install the device near heat sources (e.g. heating, drying cabinet). Do not expose the device to direct sunlight. Ensure unobstructed air circulation.
Safety Eppendorf® PlateReader AF2200 English (EN) 3.4 Information on product liability In the following cases, the designated protection of the device may be compromised. Liability for any resulting property damage or personal injury is then transferred to the operator: • The device is not used in accordance with the operating manual. • The device is used outside of its intended use. • The device is used with accessories or consumables which are not recommended by Eppendorf.
Installation Eppendorf® PlateReader AF2200 English (EN) 4 4.1 Installation Preparing installation Store the transport packaging and packing material for future safe transport or storage. Use the details included in the delivery package to check that the delivery is complete . Check all parts for any transport damage. 4.2 Removing the transport locks NOTICE! Damage to electronic components due to condensation.
Installation Eppendorf® PlateReader AF2200 English (EN) When the device is delivered, the plate compartment is secured using a transport lock to prevent damage. 1 2 2 1. Make sure the device is switched off. 2. Close the plate compartment flaps. 3. Loosen and remove the external screws (2) on the transport lock (1). 4. Remove the plate compartment by hand. 5. Loosen and remove the inner screws on the transport lock. 6. Remove the transport lock. 7. Close the plate compartment.
Installation Eppendorf® PlateReader AF2200 English (EN) 4.3 Selecting the location Find a suitable location for the device according to the following criteria: • Power supply in accordance with the name plate. • Minimum distance to other devices and walls: 10 cm. • Sufficient space for moving the plate compartment in and out. • A bench with a horizontal and even work surface that is designed to support the weight of the device (16 kg). • The surrounding area must be well ventilated.
Installation Eppendorf® PlateReader AF2200 English (EN)
Inserting the filter slide Eppendorf® PlateReader AF2200 English (EN) 5 5.1 Inserting the filter slide Information on optical filters Eppendorf offers two predefined filter slides. They are delivered preconfigured and can be used immediately for measurements. The filters are permanently installed on these predefined filter slides. They cannot be removed or exchanged. 5.1.1 Fluorescence filters The optical filters (band-pass filter) were especially developed for fluorescence measurements.
Inserting the filter slide Eppendorf® PlateReader AF2200 English (EN) 5.2 Filter slides and filter orientation 5.2.1 Filter slides The filter slides consist of an excitation section and an emission section. The filter slides allow the user to work with four independent excitation/emission filter pairs which can be defined on positions 1 to 4. The information of the inserted filters is saved on the integrated microchip.
Inserting the filter slide Eppendorf® PlateReader AF2200 English (EN) 2. Insert the filter without an arrow marking into the filter slide. The offset side of the filter with the metal lip must point away from the light source. Abb. 5-3: Filter without marking (arrow indicates the direction of the light) Fig. 5-3: 5.
Inserting the filter slide Eppendorf® PlateReader AF2200 English (EN) 5.3.2 Inserting user-specific filters The stop rings may only be used 5 times. 1. Insert the new filter into the filter slide while observing the light path. Abb. 5-5: Inserting the filter Fig. 5-5: Inserting the filter 2. Place the stop ring on the end of the filter fixing tool and turn it so it cannot fall down. 3. Push the stop ring and filter fixing tool into the filter slot. 4.
Inserting the filter slide Eppendorf® PlateReader AF2200 English (EN) 2. Adjust the filter definitions as follows: The Overview tab and four tabs for the filter definition are shown in the Filter definitions dialog window. Overview: The definition of the current filter slide is displayed for the user in the overview. Special characters (spaces, ?, $, %, ., / etc.) are not allowed in the filter slide description (exception: '_'). The filter slide description is part of the filter slide code.
Inserting the filter slide Eppendorf® PlateReader AF2200 English (EN) Wavelength: Enter the filter wavelength in the following area. (1) Fluorescence intensity mode: 230 to 600 nm (excitation) and 280 to 600 nm (emission). (2) Absorbance mode: 230 to 1000 nm. Enter the bandwidth (nm) of the filter. (3) Click on Save to adopt the new filter values. As soon as the Filter definition window is closed, the system is ready to record data with the new filters.
Inserting the filter slide Eppendorf® PlateReader AF2200 English (EN) When a filter is replaced, this information will be lost if the last flash counter reading has not been documented manually by the user. Set the counter to 0 for a completely new filter. For a filter that has already been used, enter the last recorded counter reading (if available). Optical filters may change over time depending on the frequency of use and ambient conditions. As such, they have a limited service life.
Inserting the filter slide Eppendorf® PlateReader AF2200 English (EN)
Operation Eppendorf® PlateReader AF2200 English (EN) 6 6.1 Operation Overview of operating controls 6.1.1 Notes on operation Make sure that the computer is switched on and the software is installed. The software is used for the operation and measurement. 6.1.2 Switching on the device Check to see if the device is connected to the computer via the USB cable. No initialization steps are executed when the device is switched on. 6.1.3 Starting the device The software activates the device.
Operation Eppendorf® PlateReader AF2200 English (EN) Before the start of each measurement, make sure that microtest plate position A1 is correctly inserted. Well A1 must be in the upper left position. The maximum permitted load of the plate compartment is 100 g. The maximum permitted plate height is 23 mm (including lid). 6.2 Settings The following settings are available independent of the corresponding measuring method. 6.2.
Operation Eppendorf® PlateReader AF2200 English (EN) 6.2.4 Multi mode measurements The software offers multimode measurements. This means that 10 measurements can be conducted for each plate/plate area (pattern) in the "Absorbance" or "Fluorescence intensity" modes, or in a combination of the two modes. 6.2.5 Ratio mode measurements (= measurements well by well) The well-by-well processing of one or several measurements is limited to 4 consecutive measuring strips of the same mode. 6.
Operation Eppendorf® PlateReader AF2200 English (EN) In this equation, U is the voltage, Gain is the selected amplification, 255 is the maximum amplification and 1250 V is the highest PMT voltage supply possible. Example: A gain setting of 100 corresponds to a supply voltage of 490 V: U= 6.3.3 100 * 1250 V = 490V 255 Flash settings Quick measurements with 1 flash (measurement) per well can be completed with all plate types.
Operation Eppendorf® PlateReader AF2200 English (EN) All standard microplates with 6 to 384 wells (maximum plate height: 23 mm incl. lid) that comply with the following standards can be measured: ANSI/SLAS 1-2004, ANSI/SLAS 2-2004, ANSI/SLAS 3-2004 and ANSI/SLAS 4-2004. Predefined plate definition files are installed at the same time as the operator software installation. A list with suitable microplates can be found in the software instructions. 6.4 Optimizing absorbance measurements 6.4.
Operation Eppendorf® PlateReader AF2200 English (EN) 6.4.4 MRW type The MRW type defines the pattern according to which the measurement is performed. You can select from seven different MRW types: • Square • Square (filled) • Circle • Circle (filled) • X-axis • Y-axis • XY axis Sample patterns Abb. 6-1: Square Fig. 6-1: Abb. 6-2: Square (filled) Square Abb. 6-3: Circle Fig. 6-3: Circle Fig. 6-4: Circle (filled) Abb. 6-6: Y-axis X-axis Abb. 6-7: XY axis Fig. 6-7: Square (filled) Abb.
Operation Eppendorf® PlateReader AF2200 English (EN) 6.4.5 MRW size The MRW size determines the number of points to be measured in a well. Depending on the device and the microplate type, a 1 x 1 area with a maximum of 15 x 15 points can be selected for the size. The diameter of the individual measuring points is the same as the theoretically calculated diameter of the light beam in the focus (see table).
Operation Eppendorf® PlateReader AF2200 English (EN) 6.4.7 Displaying results in Microsoft Excel A graphic, schematic overview of the measuring points is displayed in the Microsoft Excel® results sheet generated by the software. A number is assigned to each measuring point. The results are presented in a list: the measuring point number compared to the result value. The standard deviation ("Stdev") and average value ("Mean") of the measuring points/well are also displayed: Abb.
Troubleshooting Eppendorf® PlateReader AF2200 English (EN) 7 7.1 Troubleshooting General errors Error Measuring results are imprecise. Measuring results are incorrect. Positioning cannot be completed correctly. Possible cause Remedy • Reagent has exceeded its shelf life. Ensure that the reagent is still within its shelf life and properly prepared. • Reagent not prepared correctly. Use clean demineralized water of adequate quality for preparation, if required. • Pipetting incorrect.
Troubleshooting Eppendorf® PlateReader AF2200 English (EN) 7.2 Error messages If a system error occurs, the error message appears on the screen of the connected computer. System errors require an evaluation by the Technical Service. Please contact Eppendorf Service.
Maintenance Eppendorf® PlateReader AF2200 English (EN) 8 8.1 Maintenance Decontamination before shipment If you are shipping the device to the authorized Technical Service for repairs or to your authorized dealer for disposal please note the following: WARNING! Risk to health from contaminated device Follow the instructions in the decontamination certificate. It is available as a PDF file on our webpage (www.eppendorf.com/decontamination). Decontaminate all the parts you would like to dispatch.
Maintenance Eppendorf® PlateReader AF2200 English (EN) 8.3 Replacing air filters The air filter must be replaced every six months. The air filter must be replaced immediately if it is heavily contaminated. 1. Switch off the device. 2. Remove filter grille 1 and dispose of the filter insert properly. 3. Mount the filter grille with a new filter insert. 1 8.4 Cleaning DANGER! Electric shock as a result of penetration of liquid.
Maintenance Eppendorf® PlateReader AF2200 English (EN) 8.5 Disinfection/Decontamination DANGER! Electric shock as a result of penetration of liquid. Switch off the device and disconnect the power plug before starting cleaning or disinfection work. Do not allow any liquids to penetrate the inside of the housing. Do not spray clean/spray disinfect the housing. Only plug the device back in if it is completely dry, both inside and outside. 1. Remove the microplate from the device. 2.
Maintenance Eppendorf® PlateReader AF2200 English (EN) 8.6.1 Test criteria All specifications can be changed without prior notification.
Maintenance Eppendorf® PlateReader AF2200 English (EN) 8.6.2 Test instructions, upper fluorescence The following tests can be conducted on the device to check the device specifications for this measuring procedure: • Sensitivity • Precision within a plate • Precision from one measurement to the next Sensitivity Conduct the measurement with the following settings to determine the detection limit for fluorescein.
Maintenance Eppendorf® PlateReader AF2200 English (EN) Material/reagents • 1 nM fluorescein (in 0.01 M NaOH) (fluorescein sodium salt) • 0.
Maintenance Eppendorf® PlateReader AF2200 English (EN) Pipette 200 μL of 1 nM fluorescein or the blank solution (0.01 M NaOH) into the corresponding wells according to the plate layout below. X blank 1 nM fluorescein 0.01 M NaOH Material/reagents • 1 nM fluorescein (in 0.01 M NaOH) (fluorescein sodium salt) • 0.01 M NaOH (=blank solution) (NaOH pellets) • 96-well plate • 200 μL pipette with tips Calculating the precision within a plate Uniformity (%) = Mean valueF Std.
Maintenance Eppendorf® PlateReader AF2200 English (EN) Precision from one measurement to the next Carry out the following measurement to determine the precision and reproducibility.
Maintenance Eppendorf® PlateReader AF2200 English (EN) Calculating the precision from one measurement to the next Precision (CV%) = stdev wellA1 * 100 mean wellA1 Mean valueWell A1 Mean RFU value of well A1 over 20 kinetic cycles Std. deviationWell A1 Standard deviation of RFU values from well A1 over the 20 cycles The result of the formula specifies the precision from one measurement to the next in % CV (coefficient of variation). 8.6.
Maintenance Eppendorf® PlateReader AF2200 English (EN) Pipette 200 μL of 25 nM fluorescein or the blank solution (0.01 M NaOH) into the corresponding wells according to the plate layout below. X blank 25 nM fluorescein 0.01 M NaOH Material/reagents • 25 nM fluorescein (in 0.01 M NaOH) (fluorescein sodium salt) • 0.
Maintenance Eppendorf® PlateReader AF2200 English (EN) Precision across the plate Carry out the following measurement to determine the precision across the plate. Parameter Setting Measuring mode Lower fluorescence Wavelength, excitation 485 (20 nm) Wavelength, emission 535 (25 nm) Number of flashes 25 Integration time 40 (μs) Setting time 0 (μs) Signal gain Optimal Plate type GRE96fb Pipette 200 μL of 25 nM fluorescein or the blank solution (0.
Maintenance Eppendorf® PlateReader AF2200 English (EN) Calculating the precision across the plate Uniformity (%) = stdev F * 100 meanF Mean valueF Std. deviationF Mean RFU value of the wells filled with fluorophore Standard deviation of the RFU values of the wells filled with fluorophore The results of the formula specifies the precision across the plate in % CV (coefficient of variation).
Maintenance Eppendorf® PlateReader AF2200 English (EN) Pipette 200 μL of 25 nM fluorescein or the blank solution (0.01 M NaOH) into the corresponding wells according to the plate layout below. X blank 25 nM fluorescein 0.01 M NaOH Material/reagents • 25 nM fluorescein (in 0.01 M NaOH) (fluorescein sodium salt) • 0.
Maintenance Eppendorf® PlateReader AF2200 English (EN) 8.6.4 Absorbance test instructions - baseline stability Conduct the following measurements to determine the baseline stability using the available filters.
Technical data Eppendorf® PlateReader AF2200 English (EN) 9 9.1 Technical data Power supply Mains connection 100 – 120 V/220 – 240 V, 50 – 60 Hz Power consumption 150 VA Overvoltage category ll 9.2 Weight/dimensions PlateReader AF2200 16 kg Dimensions Width 462 mm (18.19") Height 282 mm (11.1") Depth 458 mm (18.03") 9.
Technical data Eppendorf® PlateReader AF2200 English (EN) 9.5 Specifications Use Universal lab device Interface USB Measurement Software-controlled Plate definitions Via software Temperature control 5 °C above ambient temperature up to 42 °C Microplates 6-well to 384-well plates (ANSI/SLAS standard formats) Shaking the plates Linear and orbital shaking, amplitude adjustable from 1 to 6 mm in 0.5 mm increments Filter handling Manually replace the filter outside of the device.
Technical data Eppendorf® PlateReader AF2200 English (EN) 9.6.1 Definition of the detection limit The detection limit is the fluorophore concentration at which the signal that is corrected by the background is equal to three times the standard deviation of the background noise. When 1 flash per well is selected, the plate compartment does not stop at the measuring position. The detection limit can be improved with more flashes per well, but the total measurement time will be longer. 9.6.
Technical data Eppendorf® PlateReader AF2200 English (EN) 9.7 Absorbance Parameters Characteristics Wavelength range 230 - 1000 nm Measuring range 0 – 4 OD The following specifications apply for a • wavelength range of 300 – 700 nm and • measurements that are conducted with 25 flashes (measurements) per well. Plate type (number of wells) 96 Accuracy 0 – 2 OD < ± 1% + 10 mOD Accuracy 2 - 3 OD < ± 2.
Ordering Information Eppendorf® PlateReader AF2200 English (EN) 10 Ordering Information Order no. (International) 6141 000.002 6141 000.010 Order no. (North America) Description 6141 000.010 PlateReader AF2200 230 V / 50 – 60 Hz 120 V / 50 – 60 Hz 6141000922 μPlate G0.5 and PlateReader AF2200 (bundle) Eppendorf microvolume plate and PlateReader AF2200 230 V / 50 – 60 Hz 120 V / 50 – 60 Hz 120 V / 50 – 60 Hz 6141 000.908 6141 000.909 6144 000.003 6144000003 μPlate G0.
Ordering Information Eppendorf® PlateReader AF2200 English (EN) Order no. (International) 0030 621.670 0030 621.700 0030 730.020 0030 741.048 0030 730.119 0030 741.013 0030 128.800 0030 128.818 Order no.
Transport, storage and disposal Eppendorf® PlateReader AF2200 English (EN) 11 11.1 Transport, storage and disposal Transport Use the original packaging for transport. Air temperature Relative humidity Atmospheric pressure General transport -25 °C – 60 °C 10 % – 95 % 30 kPa – 106 kPa Air freight -40 °C – 55 °C 10 % – 95 % 30 kPa – 106 kPa 11.2 Inserting transport locks NOTICE! Material damage due to missing transport securing device.
Transport, storage and disposal Eppendorf® PlateReader AF2200 English (EN) 11.3 Storage Air temperature Relative humidity Atmospheric pressure In transport packaging -20 °C – 60 °C 25 % – 75 % 70 kPa – 106 kPa Without transport packaging -5 °C – 45 °C 25 % – 75 % 70 kPa – 106 kPa 11.4 Disposal In case the product is to be disposed of, the relevant legal regulations are to be observed.
EG-Konformitätserklärung EC Conformity Declaration Das bezeichnete Produkt entspricht den einschlägigen grundlegenden Anforderungen der aufgeführten EG-Richtlinien und Normen. Bei einer nicht mit uns abgestimmten Änderung des Produktes oder einer nicht bestimmungsgemäßen Anwendung verliert diese Erklärung ihre Gültigkeit. The product named below fulfills the relevant fundamental requirements of the EC directives and standards listed.
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