B 6132 900.
Copyright© 2007 Eppendorf AG, Hamburg. No part of this publication may be reproduced without the prior permission of the copyright owner. Trademarks eppendorf and UVette are registered trademarks of Eppendorf AG, Hamburg, Germany. Cy is a registered trademark of GE Healthcare UK Ltd., Buckinghamshire, UK. Alexa Fluor is a registered trademark of Molecular Probes Inc., Eugene OR, USA. LabelGuard is a trademark of Implen GmbH, München, Germany.
BioPhotometer plus — Operating manual Table of contents EN Table of contents 1 User instructions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7 10 10 12 12 Preparing installation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Selecting location . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
BioPhotometer plus — Operating manual Table of contents EN 7 Maintenance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32 7.1 Operating manual 7.2 7.3 7.4 7.5 8 32 32 33 33 33 34 35 Troubleshooting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 36 8.1 8.2 8.3 9 Cleaning . . . . . . . .
BioPhotometer plus — Operating manual 1 User instructions 1 1.1 User instructions EN Using this manual Before using the device for the first time, please read this operating manual. Please view this operating manual as part of the product and keep it somewhere easily 1.2 Operating manual accessible. If this manual is lost, please request another one. The current version can be found on our website, www.eppendorf.com (International) or www.eppendorfna.com (North America).
BioPhotometer plus — Operating manual 2 Product description 2 EN Product description Main illustration Abb. 1: Front and rear view Fig. 1: Front and rear view Operating manual 2.1 2.2 1 Device display 2 Cuvette shaft cover Slide back or forward to open or close. 3 Cuvette shaft 4 Connection RS-232 5 ID plate 6 Mains connection socket 7 Fuse holder 8 Mains switch 9 Measuring keys 10 Keyboard Delivery package Number Description 2.
BioPhotometer plus — Operating manual 2 Product description The BioPhotometer plus converts the measured absorbance values into concentration results. Dependent on the method the results can be calculated via fixed factors, standards, or curve calibration. In addition to the results the device also displays the absorbance values and some other important details, e.g. the common absorbance quotients for nucleic acid calculations. Sample dilutions can also be included in the evaluation.
BioPhotometer plus — Operating manual 3 Safety 3 EN 3.1 Safety Intended use Operating manual The intended area of use for the BioPhotometer plus is the research laboratory in molecular biology, biochemistry and cell biology. The device may only be operated by trained specialist staff. The BioPhotometer plus is used to perform photometric measurements to quantify biomolecules as well as to perform turbidity measurements of microbiological cultures in routine laboratories.
BioPhotometer plus — Operating manual 3 Safety EN Warning! Risk to health from contaminated device Perform decontamination before storing or dispatching the device and/or its accessories. Operating manual WARNING! CAUTION! Caution when using aggressive chemicals. Aggressive chemicals may damage both the device and its accessories.
BioPhotometer plus — Operating manual 3 Safety EN CAUTION! Caution! Damage from improper cleaning of the cuvette shaft. Only clean the cuvette shaft using a moist cotton swab.(see Cleaning on page 32) Operating manual Do not allow any liquid to enter the cuvette shaft. Do not reach with your fingers into the cuvette shaft. CAUTION! Caution! Faulty measurement due to device confusion.
BioPhotometer plus — Operating manual 4 Installation 4 4.1 Installation EN Preparing installation Keep the transport carton and the packing material for subsequent safe transport or storage. Check the completeness of delivery based on the details of the scope of delivery (see Operating manual Delivery package on page 8). Check all parts for any transport damage. 4.
BioPhotometer plus — Operating manual 4 Installation EN 4.5 Connect printer Operating manual You can connect the Eppendorf thermal printer to the serial interface RS-232 C of the photometer (see Ordering information on page 48). 1. Connect the printer cable to the serial printer port 4 of the photometer and tighten the locking screws. 2. Connect the printer cable to the printer and also tighten the locking screws. 3.
BioPhotometer plus — Operating manual 5 Operation 5 5.1 Operation EN Overview of operating controls Control panel of the BioPhotometer plus. Fig. 3: Control panel of the BioPhotometer plus. Operating manual Abb. 3: Key Function Oval blue keys, e.g.: • In the method selection: select method group. • When entering values: enter digits. Circular keys Start standard measurement. Start blank measurement. Start sample measurement.
BioPhotometer plus — Operating manual 5 Operation EN 5.2 Methods Operating manual The following methods are available and already preprogrammed ex factory. You can modify most parameters and save them as a modified method (see Parameter on page 26). Method group Method Explanation Wavelength DNA Calculating the concentration of DNA with evaluation via factor. The methods differ mainly in the preprogrammed factor.
BioPhotometer plus — Operating manual 5 Operation 5.3 EN Summary of the measuring procedure This section contains a summary of the key steps of a measuring procedure. Operating manual 5.3.1 Prepare measurement 1. Switch on the device and, if necessary, the printer also. The device is immediately ready for measuring after being switched on. 2. Have the cuvettes for the measurements available. When selecting the cuvettes observe the respective instructions (see Cuvettes on page 13). 3.
BioPhotometer plus — Operating manual 5 Operation EN Operating manual 5.3.3 Measure Check for each measurement: • Is there enough measuring solution in the cuvette? The light path height of the BioPhotometer plus is 8.5 mm. The height of the light beam in the cuvette is 1.5 mm.
BioPhotometer plus — Operating manual 5 Operation 5.3.4 EN Finalize the method 1. Press one of the method group keys to return to the method selection and open the next method, if necessary. Operating manual 2. After all measurements are complete switch off the device. 3. Slide the cuvette shaft cover forward to protect the shaft against contamination. 5.4 Nucleic acids The methods provided in the method groups "DNA" and "RNA" differ mainly in the preprogrammed factor.
BioPhotometer plus — Operating manual 5 Operation EN 5.5 Proteins Operating manual You can measure the protein concentrations directly by measuring in the UV range at 280 nm or after adding reagent in the VIS range. 5.5.1 Protein 280 nm The measurements can be evaluated via a factor entered into the parameters or via a single point calibration (measuring a standard). • Factor: This evaluation mode is preprogrammed ex factory. However, you still have to enter the factor before the first measurement.
BioPhotometer plus — Operating manual 5 Operation 5.6 EN Methods with evaluation via standards For the following methods you can define an evaluation via calibration (measuring of standards) in the parameters: Operating manual • PROTEIN 280nm (here only calibration with one standard is possible) • BRADFORD, BRADFORD micro, BCA, BCA micro, LOWRY, LOWRY micro • Methods via the keys "assays 340 / 405 / 490" You can program up to 10 standards each in single up to triple measurement.
BioPhotometer plus — Operating manual 5 Operation EN 5.7 OD 600 Operating manual With method "OD 600" you can measure the bacteria density via a turbidity measurement at approx. 600 nm. Because this is a stray-light measurement the result depends on the geometry of the light path which can differ for the photometers by different manufacturers. The exact measurement wavelength for the BioPhotometer plus is 595 nm. Measurements with suspensions of E. coli bacteria (absorbance range: approx. 0.1 to 0.
BioPhotometer plus — Operating manual 5 Operation 5.8.3 EN Frequency of incorporation "FOI" Operating manual The FOI is a measure for the concentration of the dye relative to the concentration of the nucleic acid. It is not provided for protein methods. For calculating the FOI you can select between 2 units in the method parameters: • "MOLECULE DYE / kb": dye molecules per 1000 nucleotides. • "pmole/μg DNA": pmole dye per μg nucleic acid.
BioPhotometer plus — Operating manual 5 Operation EN Operating manual As with the nucleic acid measurement without dye measurement (see Nucleic acids on page 19) you can use the cursor keys to access additional details of the result (absorbance value and ratio 260/280) and to carry out conversions for further results: After entering the total volume of a sample: Total amount of the nucleic acid (mass in μg) and the dye (in pmole).
BioPhotometer plus — Operating manual 5 Operation EN 3. Start a sample measurement using the sample key. Operating manual 4. The result is calculated taking into account the sample dilution entered. The dilution is retained for calculating all subsequent sample results until it is overwritten. To delete the dilution enter "0" for both "SAMPLE" and "DILUENT" or delete the values using the keys clear and enter. 5.
BioPhotometer plus — Operating manual 6 Parameter and functions 6 EN Parameter and functions 6.1 Parameter Operating manual The methods of the BioPhotometer plus as well as the corresponding parameters have already been preprogrammed ex factory. You can modify these parameters. 6.1.1 View, change and store the parameters of a method 1. Select the desired method group using the corresponding method key. 2. Using the cursor keys select the desired method. 3.
BioPhotometer plus — Operating manual 6 Parameter and functions EN Entry Explanation REGRESSION Selection: Linear and non-linear regression Evaluation procedure of the multi-point calibration. • Linear regression is possible for: 2 to 10 standards • Non-linear regression is possible for: (single measurement:) 5 to 10 standards (double or triple measurement:) 4 to 10 standards. STD 1 to STD 10 Numerical input (numbers and Nominal concentration for each standard.
BioPhotometer plus — Operating manual 6 Parameter and functions EN 6.1.3 Parameters preprogrammed ex factory Operating manual Nucleic acids and OD 600 Parameter dsDNA ssDNA OLIGO DNA RNA OLIGO RNA OD 600 CUVETTE 10 mm 10 mm 10 mm 10 mm 10 mm 10 mm UNIT μg/mL μg/mL μg/mL μg/mL μg/mL MOL. UNIT pmol/mL pmol/mL pmol/μL pmol/mL pmol/μL FACTOR 50.0 37.0 30.0 40.0 30.0 CORRECTION NO NO NO NO NO CORRECTION CORRECTION CORRECTION CORRECTION CORRECTION 1.
BioPhotometer plus — Operating manual 6 Parameter and functions EN Method group "dye 550" DYE 550-dsDNA DYE 550-ssDNA DYE 550-RNA DYE 550-OLIGO DYE 550-PROTEIN CUVETTE 10 mm 10 mm 10 mm 10 mm 10 mm Unit (only for nucleic acid) μg/mL μg/mL μg/mL μg/mL mg/mL (protein: no entry, programmed permanently) MOL. UNIT (for nucleic acid) pmol/mL pmol/mL pmol/mL pmol/μL FACTOR 50.0 (for nucleic acid or protein) 37.0 40.0 30.
BioPhotometer plus — Operating manual 6 Parameter and functions EN Operating manual assay 340 Parameter ASSAY 340/1 ASSAY 340/2 ASSAY 340/3 CUVETTE 10 mm 10 mm 10 mm UNIT μg/mL μg/mL μg/mL CALCULATION Factor STD STD FACTOR ----- STD NUMBER 1-10 1 6 STD MEASUREMENT 2x 1x REGRESSION NON-LINEAR STD 1 ----- ----- STD 2 ----- STD 3 ----- STD 4 ----- STD 5 ----- STD 6 ----assay 405 and assay 490 The parameters correspond to those of method group "assay340".
BioPhotometer plus — Operating manual 6 Parameter and functions 6.2 EN Functions 1. In the method selection press the key function/·/No.. Operating manual A list of general device functions will open. In the method procedure or parameter mode this key has a different function (entering the sample number or entering a decimal point when inputting values). 2. In this function list and any sub-lists (e.g.
BioPhotometer plus — Operating manual 7 Maintenance 7 EN 7.1 Maintenance Cleaning Danger! Electric shock as a result of penetration of liquid. Operating manual Switch off the device and disconnect it from the power supply before starting cleaning or disinfecting. DANGER! Do not allow any liquids to penetrate the inside of the housing. Do not disinfect by means of spraying. Only reconnect the device to the power supply once it is completely dry.
BioPhotometer plus — Operating manual 7 Maintenance 7.2 EN Disinfection / Decontamination Danger! Electric shock as a result of penetration of liquid. Operating manual Switch off the device and disconnect it from the power supply before starting cleaning or disinfecting. DANGER! Do not allow any liquids to penetrate the inside of the housing. Do not disinfect by means of spraying. Only reconnect the device to the power supply once it is completely dry. 1.
BioPhotometer plus — Operating manual 7 Maintenance EN 7.5 Check photometer Operating manual To check the photometric accuracy and the accuracy of the wavelength Eppendorf offers a filter kit (secondary UV-VIS filter). The kit contains three filters ("sample A1", "sample A2" and "sample A3") to check the photometric acccuracy and two filters ("sample 260 nm" and "sample 280 nm") to check the accuracy of the wavelength. The absorbances of the filters are measured against a blank filter ("blank A0").
BioPhotometer plus — Operating manual 7 Maintenance 7.5.1 EN Test procedure Hint! Operating manual • Carry out test at approx. 20°C. • Remove filter only briefly from the filter box and protect against contamination or damage to the filter surface. • Protect filter against dust, heat, liquids and aggressive vapors. • Always insert the filter with the sticker containing the filter designation facing the user (towards the recipient). 1. Open the function PHOTOMETER TEST / NEW MEASUREMENT. 2.
BioPhotometer plus — Operating manual 8 Troubleshooting 8 EN Operating manual 8.1 Troubleshooting Result flags Result flags Possible cause Remedy 0.015 A340¶ Only for nucleic acid methods and None. The flag is only for information. "PROTEIN 280 nm": The marking '¶' of the A340 value indicates that the absorbance values for 230, 260 and 280 nm are corrected with the absorbance for 340 nm (parameter "CORRECTION A340" is enabled). 0.015 A550¶ or 0.
BioPhotometer plus — Operating manual 8 Troubleshooting 8.2 EN Error messages Device displays with error messages can be exited using Enter. Error text Explanation / cause Remedy (Parameter) PLEASE PROGRAM e.g.: FACTOR PLEASE PROGRAM Method parameter has not been programmed correctly or is incomplete. Check and correct parameter (see MEASURE BLANK FIRST No blank has been stored for the method opened. Measure blank. Parameter on page 26).
BioPhotometer plus — Operating manual 8 Troubleshooting EN 8.3 General errors Fault Operating manual Measuring results are imprecise. Possible cause • Shelf life of reagent solution exceeded. • Reagent not prepared correctly. • Pipetting incorrect. Remedy Ensure that the reagent is still within its shelf life and properly prepared. Use clean demineralized water of adequate quality for preparation if required.
BioPhotometer plus — Operating manual 9 Transport, storage and disposal 9 9.1 Transport, storage and disposal EN Transport Only transport the device in the original packaging. 9.2 9.3 Rel. humidity Air pressure General transportation -25 to 60°C 10 to 95% 30 to 106 kPa Air freight -40 to 55 °C 10 to 95% 30 to 106kPa Air temperature Rel.
BioPhotometer plus — Operating manual 10 Technical data 10 EN Technical data 10.1 Power supply Operating manual Power supply 100 to 240 V ± 10 % / 50 to 60 Hz ± 5 % Overvoltage category IEC 61010-1 category II Degree of contamination: IEC 61010-1 category II Power consumption approx. 20 W during operation, approx. 10 W in standby mode Permitted mains interruption approx. 10 ms at 90 V approx. 20 ms at 220 V Fuses T 1A/250 V, 5 mm x 20 mm (2 off) 10.
BioPhotometer plus — Operating manual 10 Technical data EN 10.
BioPhotometer plus — Operating manual 10 Technical data EN Operating manual 10.
BioPhotometer plus — Operating manual 11 Evaluation procedure 11 Evaluation procedure When comparing the measuring results to those of other photometers / spectrometers note that the values may depend on the bandwidths of the devices. In the following cases the differences may be significant: • The absorbance spectrum shows a narrow peak in the measurement wavelength. • The measurement is carried out not at the maximum but at the edge of a peak.
BioPhotometer plus — Operating manual 11 Evaluation procedure EN 11.2.2 Multi-point calibration: calibration line For the evaluation via a calibration line the selection "LINEAR" is programmed in the parameter "REGRESSION".
BioPhotometer plus — Operating manual 11 Evaluation procedure EN 11.4 Special evaluation procedures for the dye methods 11.4.1 Calculating the factor for the dye from the absorbance coefficient Operating manual In the dye methods a factor is calculated for the dye from the absorbance coefficient entered in the parameters and is displayed in the startup screen of the method procedure.
BioPhotometer plus — Operating manual 11 Evaluation procedure EN 11.5 Special evaluation procedures for nucleic acids and protein UV Operating manual This section relates to the evaluation of nucleic acids or proteins in the methods dsDNA, ssDNA, Oligo DNA, RNA, Oligo RNA, Protein UV and the corresponding biomolecule components in the dye methods. 11.5.1 Correction A340 Application: Partial correction of distortions of absorbance due to turbidity in the measuring solution.
BioPhotometer plus — Operating manual 11 Evaluation procedure EN Calculation of the molar concentration Operating manual Application: calculation of the molar concentration of the nucleic acid from the mass concentration and relative molar mass. The molar mass is either entered directly or calculated by the device from the entered figure of the bases or base pairs per nucleic acid molecule.
BioPhotometer plus — Operating manual 12 Ordering information EN Operating manual 12 Ordering information Order No. (International) Order No. (North Description America) 6132 000.008 6132 000.016 952000006 BioPhotometer plus 230 V / 50-60 Hz, power plug Europe, more types of mains connection available 120 V / 50-60 Hz, power plug North America 952010221 Secondary UV-VIS-Filter Test filter set for checking photometric accuracy und wavelength accuracy (according to NIST) 6131 011.006 6131 010.
In touch with life Your local distributor: www.eppendorf.com/worldwide Eppendorf AG · 22331 Hamburg · Germany · Tel: +49 40 53801-0 · Fax: +49 40 538 01-556 · E-mail: eppendorf@eppendorf.com Eppendorf North America, Inc. · 102 Motor Parkway · Hauppauge, N.Y. 11788-5178 · USA Tel: +1 516 334 7500 · Toll free phone: +1 800-645-3050 · Fax: +1 516 334 7506 · E-mail: info@eppendorf.com Application Support Europe: Tel: +49 1803 666 789 (Preis je nach Tarif im Ausland; 9 ct/min aus dem dt.
