Manual

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BioPhotometer

Bedienungsanleitung

Operating Manual

Mode d'emploi

Istruzioni d'impiego

Manual de Instrucciones

BP_Titel_d_e_f_i_sp.fm Seite 1 Montag, 20. Februar 2006 10:23 Uhr

Summary of content (53 pages)

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BP_Titel_d_e_f_i_sp.fm Seite 1 Montag, 20.
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BP_Titel_d_e_f_i_sp.fm Seite 4 Montag, 20.
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Inhalt_d_e_f_i_sp_Copy.fm Seite 1 Montag, 20. Februar 2006 10:25 Uhr Bedienungsanleitung . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 Operating Manual . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47 Mode d’emploi . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
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Inhalt_d_e_f_i_sp_Copy.fm Seite 2 Montag, 20.
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00_Inhalt_e.fm Seite 47 Montag, 20. Februar 2006 10:46 Uhr Contents 1 Overview . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 49 2 Technical data . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 51 3 Safety precautions and prevention of damage . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 53 4 4.1 4.2 4.
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00_Inhalt_e.fm Seite 48 Montag, 20.
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01_Ueberblick_e.fm Seite 49 Montag, 20. Februar 2006 10:47 Uhr 1 Overview 1 8 2 7 6 3 1 2 3 4 4 5 Device display 9 method keys Function key (device functions) Parameter key (programming key) 5 6 7 8 Dilution key Conversion key Measuring keys Cuvette shaft The main power key, main power connection and printer connection are located on the rear of the device (see Section 4, "Installation").
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01_Ueberblick_e.fm Seite 50 Montag, 20.
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02_TecDat_e.fm Seite 51 Montag, 20.
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02_TecDat_e.fm Seite 52 Montag, 20. Februar 2006 16:06 Uhr 2 Technical data Operation Cuvette material: dsDNA, ssDNA, RNA, Oligo, Protein: Quartz glass or plastic (UVette® from Eppendorf) OD 600, Bradford, Lowry, BCA: Glass or plastic Cuvette shaft: 12.5 mm x 12.5 mm, not temperature-controlled Overall height of cuvettes: Min. 36 mm Height of light beams in the cuvette: 8.5 mm Light bundle in the cuvette: Width: 1 mm Height: 1.
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03_Sicher_e.fm Seite 53 Montag, 20. Februar 2006 10:49 Uhr 3 Safety precautions and prevention of damage Before using the Biophotometer please familiarize yourself completely with the operating instructions. The following points must be followed exactly to enable safe work with the device: Technical safety – Do not open the device. – Do not allow any liquid to enter into the device. – Disconnect the device from the mains supply before carrying out maintenance work or changing the fuses.
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04_Installation_e.fm Seite 54 Montag, 20. Februar 2006 10:50 Uhr 4 Installation Delivery package – – – – BioPhotometer Mains cable for BioPhotometer Operating manual, incl. short instructions Seal for cuvette shaft 4.1 BioPhotometer Connect up device Space required: Width: Depth: 40 cm 50 cm Power connection: Safety socket – Insert the mains plug of the device into the safety socket.
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04_Installation_e.fm Seite 55 Montag, 20. Februar 2006 10:50 Uhr 4 Installation 4.2 Printer Printer DPU 414 The Eppendorf Thermal Printer DPU 414 can be connected to the serial interface RS-232 C of the BioPhotometer (see Section 11, "Ordering information"). – Insert the printer cable into the printer connection socket of the BioPhotometer (see photo) and tighten the safety screws on the plug to secure. – Connect the printer cable to the printer and tighten the safety screws on the plug to secure.
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04_Installation_e.fm Seite 56 Montag, 20. Februar 2006 10:50 Uhr 4 Installation Other printers In addition to the DPU 414, it is also possible to connect other serial printers to the serial interface of the BioPhotometer. With the aid of an adapter cable, parallel printers can also be connected. BioPhotometer – Select the function "Printer serial" in the functions list, and conﬁrm.
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05_Bedienung_e.fm Seite 57 Dienstag, 21. Februar 2006 10:09 Uhr 5 Operation 5.1 Keypad Photometer Standard 7 dsDNA 8 ssDNA 9 RNA 4 Protein 5 OD 600 6 Oligo 1 Bradford 2 Lowry 3 BCA 0 Sample No. • Function Parameter Blank Sample Conversion Dilution 7 dsDNA – To call up the "Double-stranded DNA" method. – To enter ﬁgure 7. 8 ssDNA – To call up the "Single-stranded DNA" method. – To enter ﬁgure 8. 9 RNA – To call up the "RNA" method. – To enter ﬁgure 9.
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05_Bedienung_e.fm Seite 58 Dienstag, 21. Februar 2006 10:09 Uhr 5 Operation 3 BCA 5 OD 600 Parameter • Function 0 Sample No. Dilution Conversion Clear Enter Standard Blank Sample 58 – To call up the "BCA" and "BCA micro" methods. – To switch between the "BCA" and "BCA micro" methods. – To enter ﬁgure 3. – To call up the "OD 600 (measuring the bacteria density)" method. – To enter ﬁgure 5. – To call up the programming level. – To exit the programming level. – To call up the function level.
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05_Bedienung_e.fm Seite 59 Dienstag, 21. Februar 2006 10:09 Uhr 5 Operation 5.2 Measuring nucleic acids This description is valid for the following methods: – dsDNA – ssDNA – RNA – Oligo Call up method 7 dsDNA dsDNA P RO G R A M M E D FAC TO R : 1 A 260 = 5 0 . 0 µg/mL Blank or Sample Calculation The factory-set factors are those which are normally used with nucleic-acid methods for the conversion of UV absorbance into concentration (in this example: 50).
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05_Bedienung_e.fm Seite 60 Dienstag, 21. Februar 2006 10:09 Uhr 5 Operation Measure sample Sample dsDNA 70.0 1.97 2.03 260/ 280 260/ 230 SAMPLE 001 µg/mL 0.694 1.408 0.715 0.002 A 230 A260 A 280 A 320 Results display As an indication of the purity of the nucleic acid sample which has been measured, the absorbance at 230, 280 and 320 nm as well as the ratios A260/A280 and A260/A230 are displayed in addition to the concentration result and the absorbance at a wavelength of 260 nm.
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05_Bedienung_e.fm Seite 61 Dienstag, 21. Februar 2006 10:09 Uhr 5 Operation 1 Bradford Display after entry of "140 µL sample volume" and "300 base pairs": 3 BCA 4 Protein 0 Sample No. 0 Sample No. 0 Sample No. Enter dsDNA SAMPLE 001 70.0 µg/mL 9.8 µg 353.5 pmol/mL 49.5 pmol Enter Enter The molar unit of concentration (here: "pmol/mL") is preprogrammed, but can be selected and changed using the key. Parameter 5.
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05_Bedienung_e.fm Seite 62 Dienstag, 21. Februar 2006 10:09 Uhr 5 Operation Measuring procedure The following example shows the measuring procedure for the "Absorbance" calculation mode. For details of the measuring procedure via standard (one-point calibration), please refer to "Measuring proteins with reagent". Blank measurements remain stored until the date changes (For further details, please refer to "Measuring nucleic acids"). Measure blank Measure sample Blank Sample P ROT E I N BLANK 0.
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05_Bedienung_e.fm Seite 63 Dienstag, 21. Februar 2006 10:09 Uhr 5 Operation 5.4 Measuring proteins with reagent (Bradford, BCA, Lowry) Call up method 1 Bradford BRADFORD C A L I B R AT I O N R A N G E XXX – XXX µg/mL Blank or Standard If a valid calibration (which is then stored by the device) has already been performed, the date and time of the stored calibration appear.
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05_Bedienung_e.fm Seite 64 Dienstag, 21. Februar 2006 10:09 Uhr 5 Operation Measuring procedure Blank measurements remain stored until the date changes (For further details, see "Measuring nucleic acids"). Standard measurements remain stored until they are overwritten with new standard measurements. For the calculation of sample measurements, the most-recently stored calibration is used.
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05_Bedienung_e.fm Seite 65 Dienstag, 21. Februar 2006 10:09 Uhr 5 Operation Device display following all standard measurements: BRADFORD X.XXX STD 5–2 XXXX µg/mL A C V: 2 . 8 % C A L I B R AT I O N S TO R E D The CV (coefﬁcient of variation) is a measure of the scattering of standard values around the regression curve. If the CV is smaller than 10 %, the calibration is stored automatically.
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05_Bedienung_e.fm Seite 66 Dienstag, 21. Februar 2006 10:09 Uhr 5 Operation 5.5 Measuring OD 600 Select method 5 OD 600 OD600 Blank or Sample Measuring procedure Blank measurements remain stored until the date changes (for further details, see "Measuring nucleic acids"). Measure blank Measure sample Blank Sample OD600 BLANK 0.000 A Blank or OD600 0.
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05_Bedienung_e.fm Seite 67 Dienstag, 21. Februar 2006 10:09 Uhr 5 Operation 5.6 Measuring diluted samples Sample dilutions may be entered using the Dilution key before the measurement begins. When the result is calculated and displayed, the dilution factor is included automatically. In the following example, a blank has already been measured: dsDNA Enter dilution BLANK 0.000 A Blank or dsDNA Sample SAMPLE 001 Dilution SAMPLE+DILUENT – – – + – – – – µL 2 Lowry dsDNA 1 Bradford 0 Sample No.
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05_Bedienung_e.fm Seite 68 Dienstag, 21. Februar 2006 10:09 Uhr 5 Operation 5.7 Changing the sample number During sample measurements, the serial number of the sample appears in the top right of the display. The sample number is counted separately for each method and is reset to "1" when the date changes. The sample number can be changed as desired (e.g. for repeat measurements): dsDNA 70 .0 SAMPLE 005 µg/mL 2+180µL 1.97 2.03 Change sample number 0 Sample No. 3 BCA 0.694 1.408 0.715 0.
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06_Programm_e.fm Seite 69 Montag, 20. Februar 2006 10:54 Uhr 6 Programming 6.1 Programming procedure For each method, parameters such as the type of calculation or the unit of concentration are stored. The factory-set method programs can be changed using the Parameter key. Call up method OLIGO 6 Oligo P RO G R A M M E D FAC TO R : 1 A 260 = 3 0 . 0 µg/mL Blank Call up parameter list OLIGO Sample or PAG E 1 – 3 Parameter ▲ FAC TO R 30.0 C O R R . W I T H A 320 . . . . . . . . . . . .
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06_Programm_e.fm Seite 70 Montag, 20. Februar 2006 10:54 Uhr 6 Programming Example: Changing the unit Selection parameters are selected using the cursor keys and conﬁrmed by pressing Enter . The stored setting is marked with an asterisk (*): Select parameter OLIGO Dilution PAG E 2 – 3 µg/mL * ng/µL – µg/µL – M. UNIT . . . . . . . . . . pmol/µL * µmol/L – ▲ UNIT . . . . . . . . . . . . . . . . . Store parameter OLIGO PAG E 2 – 3 Enter µg/mL – ng/µL – µg/µL * UNIT . . . . . . . . . . . . . . .
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06_Programm_e.fm Seite 71 Montag, 20. Februar 2006 10:54 Uhr 6 Programming 6.2 Overview of parameters dsDNA ssDNA RNA Oligo Protein Bradford Brad.micro Lowry Low.
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06_Programm_e.fm Seite 72 Montag, 20. Februar 2006 10:54 Uhr 6 Programming 6.3 Explanation of parameters Parameters are deﬁned as selection parameters or as parameters for entering numbers. In the case of selection parameters, the programmable alternatives are method-dependent (see overview in previous section). Entries Explanation Calculation Selection Selection of calculation procedures: Absorbance, Factor, Standard and Warburg formula.
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06_Programm_e.fm Seite 73 Montag, 20. Februar 2006 10:54 Uhr 6 Programming The following parameters are offered only when the "Standard" calculation procedure has been programmed: Parameter Entries Explanation Std. number Entry of numbers ("1" to "10") Number of different standards. Std. measurement Selection Selection from "1x", "2x", "3x" repeat measurement of each standard; a mean value is formed for the further calculation using the repeat measurements.
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74 Notes: 1) 2) 3) 4) 37.0 off µg/mL pmol/mL 10 mm 50.0 off µg/mL pmol/mL 10 mm ssDNA 10 mm µg/mL pmol/mL 40.0 off RNA 10 mm µg/µL pmol/µL 30.0 off Oligo Protein 6 1x 6 1x 10 mm 100 250 500 750 1000 1500 - - - - - 4) 10 mm µg/mL non-linear - - - - - 1) µg/mL3) 1x2) - - - - - 1) off 6 1x Standard - - - - - 1) Lowry 6 1x Lowry micro Standard - - - - - 1) 8 1x Standard - - - - - 1) BCA 5 1x 10 mm 1.00 2.
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07_Funktion_e.fm Seite 75 Montag, 20. Februar 2006 10:55 Uhr 7 Functions Functions list Function Entries Display results Call up using Explanation Enter . Display of the last 100 results (The most-recent result appears ﬁrst): Dilution Conversion Calibration report Call up using Enter . Enter : • Function : : To select the results. To print out the results that have just been displayed. To return to the functions list.
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07_Funktion_e.fm Seite 76 Montag, 20. Februar 2006 10:55 Uhr 7 Functions Example: Changing the language version Call up functions list Dilution FUNKTION SEITE 1–4 ▲ Select desired function • Function ERGEBNISSE ANZEIGEN K A L I B R AT I O N S R E P O RT DAT U M UHRZEIT 27.06.1998 20:44 FUNKTION SEITE 3–4 ▲ S P R AC H E D E U T S C H * L A N G UAG E E N G L I S H – L A N G UAG E U. S.
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08_Fehler_e.fm Seite 77 Montag, 20. Februar 2006 11:24 Uhr 8 Error messages, result ﬂagging and help texts Result ﬂagging 1.586 A260 ▲ Explanation Flagging of A260 in the display or on the printout (for method "Protein direct" only): The method was calculated using the Warburg formula. 0.
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08_Fehler_e.fm Seite 78 Montag, 20. Februar 2006 11:24 Uhr 8 Error messages, result ﬂagging and help texts Error texts in measuring procedure Error text Explanation / Cause Solution Measure blank ﬁrst No blank has been measured for the method selected. Measure the blank. Measure standard ﬁrst No valid calibration for the method selected. not within calibration Measurement module Error 1 Measurement module Error 2 Measurement module Error 3 – Measure standards.
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08_Fehler_e.fm Seite 79 Montag, 20. Februar 2006 11:24 Uhr 8 Error messages, result ﬂagging and help texts Error text Explanation / Cause Solution CV greater than 10 % (Following standard measurements:) The scattering of the measured values around the calculated calibration line or curve is very large (see Section 12, "Calculation"). Check calibration result. – – Enter : Store calibration. : Abort calibration. Recalibrate or use the calibration stored.
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09_Wartung_e.fm Seite 80 Montag, 20. Februar 2006 11:25 Uhr 9 Maintenance and cleaning Photometer – Disconnect the device from the main power source before carrying out maintenance work or to change the fuses. The inside of the device is a high-voltage area. Danger! – Wipe the entire device using a moist cloth and a mild cleaning agent. – Disinfect the device using a lightly moistened cloth and a 70 % ethanol/water mixture. – Do not allow any liquid to enter the device.
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10_Kurz_e.fm Seite 81 Montag, 20. Februar 2006 11:33 Uhr 10 Short instructions Preparation The BioPhotometer is ready to measure immediately after being switched on. Methods 7 dsDNA 8 ssDNA 9 RNA 8 ssDNA 9 RNA 4 Protein 5 OD 600 6 Oligo 1 Bradford 2 Lowry 3 BCA dsDNA ssDNA RNA Oligo – Direct measurement of the nucleic acids at 260 nm. – Ratios A260/A280 and A260/A230. – Optional correction of absorbance values via A320. – Measurement using quartz-glass cuvette or UVette® from Eppendorf.
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10_Kurz_e.fm Seite 82 Montag, 20. Februar 2006 11:33 Uhr 10 Short instructions Programming The factory-set method programs may be changed as required. Example: Programming of the unit "µg/mL" and of calculation via standard (500 µg/mL) for the Protein method.
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10_Kurz_e.fm Seite 83 Montag, 20. Februar 2006 11:33 Uhr 10 Short instructions Measuring procedure for dsDNA dsDNA P RO G R A M M E D FAC TO R 1 A260 = 50.0 µg/mL 7 dsDNA Select dsDNA method Blank or dsDNA Measure blank Blank When the sample is diluted: Example: 20 + 200 µL dsDNA Dilution 2 Lowry 0 Sample No. Enter 2 Lowry 0 Sample No. 0 Sample No. Enter BLANK 0.000 Blank A or Sample SAMPLE 001 20+200 µL Blank or Sample dsDNA SAMPLE 001 563.
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10_Kurz_e.fm Seite 84 Montag, 20. Februar 2006 11:33 Uhr 10 Short instructions Bradford measuring procedure BRADFORD C A L I B R AT I O N R A N G E 100 – 2000 µg/mL 1 Bradford Select Bradford method or Blank If "Bradford micro" is to be selected: Standard B R A D. m i c r o C A L I B R AT I O N R A N G E 1.0 – 20.0 µg/mL 1 Bradford Blank or Standard BRADFORD Measure blank BLANK 0.000 Blank A or Sample Blank BRADFORD Measure ﬁrst standard 0.
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11_Bestell_e.fm Seite 85 Montag, 20. Februar 2006 11:32 Uhr 11 Ordering information 11.1 Ordering information Order no. 6131 000.012 Photometer BioPhotometer (230 V; 50/60 Hz; European plug) (additional power supply variants available) 6131 810.006 BioPhotometer Software Package for online data transfer on PC 6131 928.007 Secondary UV-VIS ﬁlter, Test Filter Set, for checking the BioPhotometer 6131 011.006 Printer Thermal Printer DPU 414, incl. power supply 230 V unit and printer cable 0013 021.
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12_Auswertg_e.fm Seite 86 Montag, 20. Februar 2006 11:34 Uhr 12 Calculation 12.1 Nucleic acids (dsDNA, ssDNA, RNA, oligo) Calculation via factor C = A260 x F C = Calculated concentration A260 = Absorbance measured at 260 nm F = Factor (method-speciﬁc programming using the Parameter key) The nucleic acid methods have the following special feature: The programmed factor is always based on the unit of concentration "µg/mL".
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12_Auswertg_e.fm Seite 87 Montag, 20. Februar 2006 11:34 Uhr 12 Calculation Conversion key: Calculating the molar concentration Application: Calculating the molar concentration from the mass concentration and the relative molar mass. The molar mass is either entered directly or calculated by the device using the number of bases / base pairs per molecule.
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12_Auswertg_e.fm Seite 88 Montag, 20. Februar 2006 11:34 Uhr 12 Calculation Sample dilution, optical light path of the cuvette and correction A320 See Section 12.1. 12.
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12_Auswertg_e.fm Seite 89 Montag, 20. Februar 2006 11:34 Uhr 12 Calculation Sample dilution and optical light path of the cuvette See Section 12.1. 12.4 OD 600 The measured values appear as absorbance values measured at a wavelength of 595 nm. Sample dilution and optical light path of the cuvette See Section 12.1.
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13_Ueberpruef_e.fm Seite 90 Montag, 20. Februar 2006 11:35 Uhr 13 Testing the photometer To enable the photometric accuracy and the wavelength accuracy to be tested, a ﬁlter set (secondary UV-VIS ﬁlter) is available from Eppendorf. This set contains three ﬁlters ("Sample A1", "Sample A2" and "Sample A3") for testing the photometric accuracy and two ﬁlters ("Sample 260 nm" and "Sample 280 nm") for testing the wavelength accuracy. The absorbance of the ﬁlters is measured against a blank ﬁlter ("Blank A0").
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13_Ueberpruef_e.fm Seite 91 Montag, 20. Februar 2006 11:35 Uhr 13 Testing the photometer Test procedure – Carry out the test at approximately 20 °C. – Remove the ﬁlter from the ﬁlter box for a brief period only. Make sure that the surface of the ﬁlter is not contaminated or damaged. – Protect the ﬁlter from dust, heat, liquid and aggressive vapors. – When inserting the ﬁlter, ensure that the label with the ﬁlter description is facing the user. – Select the function "Photometer test".
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BioPh_Conformity_e.fm Seite 92 Montag, 20. Februar 2006 11:36 Uhr Conformity Declaration for BioPhotometer 6131 in accordance with enclosure 15 of "Eichordnung" (German standardization regulations) Description of measurement Device used: Type: Manufacturer / Distributor: Mode of instruction: 1.
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03_Kurz_sp.fm Seite 153 Montag, 20. Februar 2006 14:56 Uhr 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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BP_Titel_d_e_f_i_sp.fm Seite 3 Montag, 20.
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BP_Titel_d_e_f_i_sp.fm Seite 2 Montag, 20. Februar 2006 10:23 Uhr Your local distributor: www.eppendorf.com/worldwide Eppendorf AG 22331 Hamburg · Germany Tel. +49 40 538 01-0 Fax +49 40 538 01-556 E-Mail: eppendorf@eppendorf.com Eppendorf North America, Inc. One Cantiague Road, P.O. Box 1019 Westbury, N.Y. 11590-0207 USA Tel. +1 516 334 7500 Toll free phone 800 645 3050 Fax +1 516 334 7506 E-Mail: info@eppendorf.com Application Support Europe, International: Tel.