Eppendorf BioSpectrometer fluorescence Manual do Utilizador

nualoSpectrometer® fluorescenceEN)g manual Register your instrument! www.eppendorf.com/myeppendorfEppendorf BioSpectrometer® fluorescenceOperating ma

Operating instructionsEppendorf BioSpectrometer® fluorescenceEnglish (EN)10

Evaluate your manualGive us your feedback.www.eppendorf.com/manualfeedbackYour local distributor: www.eppendorf.com/contactEppendorf AG · 22331 Hambur

11Product descriptionEppendorf BioSpectrometer® fluorescenceEnglish (EN)2 Product description2.1 Main illustrationAbb. 2-1: Front and rear viewFig. 2-

Product descriptionEppendorf BioSpectrometer® fluorescenceEnglish (EN)122.3 FeaturesThe BioSpectrometer fluorescence combines two spectroscopic measur

13SafetyEppendorf BioSpectrometer® fluorescenceEnglish (EN)3Safety3.1 Intended useThe BioSpectrometer fluorescence is to be used in molecular biology,

SafetyEppendorf BioSpectrometer® fluorescenceEnglish (EN)14WARNING! Electric shock due to damage to device or mains cable. Only switch on the device

15SafetyEppendorf BioSpectrometer® fluorescenceEnglish (EN)3.3.2 Damage to deviceNOTICE! Damage from the use of aggressive chemicals. Do not use any

SafetyEppendorf BioSpectrometer® fluorescenceEnglish (EN)163.4 Information on product liabilityIn the following cases, the designated protection of th

17InstallationEppendorf BioSpectrometer® fluorescenceEnglish (EN)4 Installation4.1 Preparing installation Keep the transport carton and the packing m

InstallationEppendorf BioSpectrometer® fluorescenceEnglish (EN)184.4 Connecting the printer4.4.1 Thermal printer DPU-S445PrerequisitesSoftware version

19InstallationEppendorf BioSpectrometer® fluorescenceEnglish (EN)4.5 Connecting PC or USB stick for data exportYou can connect a FAT 32-formatted USB

6137 900.058-02/052014Copyright © 2014 Eppendorf AG, Hamburg. No part of this publication may be reproduced without the prior permission of the copyr

InstallationEppendorf BioSpectrometer® fluorescenceEnglish (EN)20

21OperationEppendorf BioSpectrometer® fluorescenceEnglish (EN)5 Operation5.1 Overview of operating controlsAbb. 5-1: Control panel of the BioSpect rom

OperationEppendorf BioSpectrometer® fluorescenceEnglish (EN)22Move the cursor to the left, right, up, down.• Navigation between input fields.• and

23OperationEppendorf BioSpectrometer® fluorescenceEnglish (EN)5.1.1 Entering textYou can enter texts when assigning method names and result units. Res

OperationEppendorf BioSpectrometer® fluorescenceEnglish (EN)245.2 Inserting the cuvetteStandard rectangular glass or plastic cuvettes can be inserted

25OperationEppendorf BioSpectrometer® fluorescenceEnglish (EN)1. Position the cuvette so that the optical window of the cuvette is pointing towards th

OperationEppendorf BioSpectrometer® fluorescenceEnglish (EN)265.3.2 Measuring procedure5.3.2.1 Selecting a method5.3.2.2 Checking parameters Use the

27OperationEppendorf BioSpectrometer® fluorescenceEnglish (EN)5.3.2.3 Measuring the blank and standards5.3.2.4 Measuring samplesFor evaluations withou

OperationEppendorf BioSpectrometer® fluorescenceEnglish (EN)285.3.2.5 Finalizing the method5.3.2.6 Optional: process results5.3.2.7 Printing and expor

29OperationEppendorf BioSpectrometer® fluorescenceEnglish (EN)5.3.3 Important measurement instructionsCheck for each measurement:• For plastic cuvette

3Table of contentsEppendorf BioSpectrometer® fluorescenceEnglish (EN)Table of contents1 Operating instructions . . . . . . . . . . . . . . . . . . . .

OperationEppendorf BioSpectrometer® fluorescenceEnglish (EN)30

31MethodsEppendorf BioSpectrometer® fluorescenceEnglish (EN)6 Methods6.1 Selecting a methodMethods and method templates are delivered preprogrammed. T

MethodsEppendorf BioSpectrometer® fluorescenceEnglish (EN)32Tab. 6-2: Fluorimetric methodsYou can create new methods in all folders using <New Meth

33MethodsEppendorf BioSpectrometer® fluorescenceEnglish (EN)6.2.2 Routine method groupThe methods for the Routine group are preprogrammed as fixed met

MethodsEppendorf BioSpectrometer® fluorescenceEnglish (EN)34• Correction of the influence of the dye spectrum on the accuracy of the biomolecule measu

35MethodsEppendorf BioSpectrometer® fluorescenceEnglish (EN)6.3 Fluorimetry method description6.3.1 Routine method groupThe methods for the Routine gr

MethodsEppendorf BioSpectrometer® fluorescenceEnglish (EN)36Calibration curve• Measurement of the RFU values and evaluation with a series of 2 to 12 s

37MethodsEppendorf BioSpectrometer® fluorescenceEnglish (EN)Unit Selection:mg/mL | μg/mL | ng/mL | pg/mL | μg/μL | mg/dL | μmol/mL | nmol/mL | pmol/mL

MethodsEppendorf BioSpectrometer® fluorescenceEnglish (EN)38Factor Value input:Factor.Limit: max. of 6 digits including decimal point.Factor for conve

39MethodsEppendorf BioSpectrometer® fluorescenceEnglish (EN)Correct A260 1 Selection:on | offOnly for the Dye labels method group.Correction of the in

Table of contentsEppendorf BioSpectrometer® fluorescenceEnglish (EN)46 Methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

MethodsEppendorf BioSpectrometer® fluorescenceEnglish (EN)406.5 Method procedureFOI Selection:none | dye/kb | pmole/μgOnly for the Dye labels method g

41MethodsEppendorf BioSpectrometer® fluorescenceEnglish (EN)A method procedure is composed of a maximum of 5 steps. The currently active step is highl

MethodsEppendorf BioSpectrometer® fluorescenceEnglish (EN)426.5.2 Measure standardsSaving the method under a new name:You can save the method in the s

43MethodsEppendorf BioSpectrometer® fluorescenceEnglish (EN)6.5.3 Measure samplesThe sample key is used for measuring your samples consecutively. Blan

MethodsEppendorf BioSpectrometer® fluorescenceEnglish (EN)44Enter dilutionSoftkeys• [Clear dil.]: Delete values for sample dilution.• [OK]: Confirm sa

45MethodsEppendorf BioSpectrometer® fluorescenceEnglish (EN)Softkeys• [Keyboard]: Display keyboard.• [abc]: Change between upper and lower case letter

MethodsEppendorf BioSpectrometer® fluorescenceEnglish (EN)466.5.4 Measure samples: Results displaysThis section contains a display of typical results

47MethodsEppendorf BioSpectrometer® fluorescenceEnglish (EN)Routine main groupNucleic acids Results display:• Concentration result with absorbance at

MethodsEppendorf BioSpectrometer® fluorescenceEnglish (EN)48Proteins (with reagent)Results display:• Concentration result with absorbance at the measu

49MethodsEppendorf BioSpectrometer® fluorescenceEnglish (EN)Basic main groupFactor, standard Analog to Protein direct UV (see above) Results display:•

5Table of contentsEppendorf BioSpectrometer® fluorescenceEnglish (EN)11 Technical data. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

MethodsEppendorf BioSpectrometer® fluorescenceEnglish (EN)50Proteins Analog to Nucleic acids (see above). Results display:• Concentration result with

51MethodsEppendorf BioSpectrometer® fluorescenceEnglish (EN)6.5.5 Process resultsThe sample measurement is followed by two optional steps in the metho

MethodsEppendorf BioSpectrometer® fluorescenceEnglish (EN)526.5.6 Process results: OptionsZoomPress the [Zoom] softkey and select one of the following

53MethodsEppendorf BioSpectrometer® fluorescenceEnglish (EN)For all 3 versions, the [reset zoom] softkey will bring you back to the original display o

MethodsEppendorf BioSpectrometer® fluorescenceEnglish (EN)54Peak detectionPress the [Peaks] softkey. For the peak detection you can alternate between

55MethodsEppendorf BioSpectrometer® fluorescenceEnglish (EN)6.5.7 Print & exportIn the last optional method step, you can assemble data packets fo

MethodsEppendorf BioSpectrometer® fluorescenceEnglish (EN)56Data exportThe data will be transferred as Excel files (.xls) and can be read with Excel v

57MethodsEppendorf BioSpectrometer® fluorescenceEnglish (EN)Select data packetsBased on the method and parameter setting, only the available data pack

MethodsEppendorf BioSpectrometer® fluorescenceEnglish (EN)58

59FunctionsEppendorf BioSpectrometer® fluorescenceEnglish (EN)7Functions7.1 Functions of the User main groupWith the function key or the [Function] so

Table of contentsEppendorf BioSpectrometer® fluorescenceEnglish (EN)6

FunctionsEppendorf BioSpectrometer® fluorescenceEnglish (EN)607.1.1 Results memoryDevice calibration • Option for checking the spectrophotometer; An E

61FunctionsEppendorf BioSpectrometer® fluorescenceEnglish (EN)7.1.2 General method parameters If you would like to print or export results, select th

FunctionsEppendorf BioSpectrometer® fluorescenceEnglish (EN)62When programming a method of the Dye labels or Proteins direct UV method groups, you can

63FunctionsEppendorf BioSpectrometer® fluorescenceEnglish (EN)7.1.3 Absorbance spectra library•NA name•Factor• Double-strandedThe factor is used to ca

FunctionsEppendorf BioSpectrometer® fluorescenceEnglish (EN)647.1.4 Device settings7.1.5 Device calibrationInformation on checking the device is provi

65MaintenanceEppendorf BioSpectrometer® fluorescenceEnglish (EN)8 Maintenance8.1 Cleaning1. Wipe down the surfaces with a cloth moistened with a mild

MaintenanceEppendorf BioSpectrometer® fluorescenceEnglish (EN)668.1.1 Cleaning the cuvette shaft coverIf you would like not only to clean the directly

67MaintenanceEppendorf BioSpectrometer® fluorescenceEnglish (EN)5. If the cuvette shaft cover needs to be removed for the disinfection, proceed as fol

MaintenanceEppendorf BioSpectrometer® fluorescenceEnglish (EN)68Please connect the Eppendorf thermal printer if you would like to document the values.

69MaintenanceEppendorf BioSpectrometer® fluorescenceEnglish (EN)8.3.1.1 Checking photometric accuracy1. Select the Spectrometer unit function in the D

7Operating instructionsEppendorf BioSpectrometer® fluorescenceEnglish (EN)1 Operating instructions1.1 Using this manual Read this operating manual co

MaintenanceEppendorf BioSpectrometer® fluorescenceEnglish (EN)70Have the filter recertified by Eppendorf Service after 2 years.8.3.1.2 Checking wavele

71MaintenanceEppendorf BioSpectrometer® fluorescenceEnglish (EN)8.3.2 Checking the fluorescence unit8.3.3 Device self-testThe frequency of the automat

MaintenanceEppendorf BioSpectrometer® fluorescenceEnglish (EN)72• Determination of the random error of the excitation light ("noise")• Deter

73TroubleshootingEppendorf BioSpectrometer® fluorescenceEnglish (EN)9 Troubleshooting9.1 General errorsError Possible cause RemedyMeasuring results ar

TroubleshootingEppendorf BioSpectrometer® fluorescenceEnglish (EN)74The measuring results are not correct.• The method has not been programmed correct

75TroubleshootingEppendorf BioSpectrometer® fluorescenceEnglish (EN)9.2 Error messagesYou can exit device displays with error messages using the [OK]

TroubleshootingEppendorf BioSpectrometer® fluorescenceEnglish (EN)76A method (or folder, dye, protein, nucleic acid, or unit) with this name already e

77TroubleshootingEppendorf BioSpectrometer® fluorescenceEnglish (EN)The ID cannot be set. • Error when entering the sample ID. Different causes are po

TroubleshootingEppendorf BioSpectrometer® fluorescenceEnglish (EN)789.3 Result flagsWarnings and error messages for results are displayed in the botto

79TroubleshootingEppendorf BioSpectrometer® fluorescenceEnglish (EN)The coefficient of determination is <0.8.• For methods with evaluation of stand

Operating instructionsEppendorf BioSpectrometer® fluorescenceEnglish (EN)81.3 Symbols usedDepiction Meaning1.2.Actions in the specified order Action

TroubleshootingEppendorf BioSpectrometer® fluorescenceEnglish (EN)80At least one of the results is negative.• For methods with several results (e.g.,

81Transport, storage and disposalEppendorf BioSpectrometer® fluorescenceEnglish (EN)10 Transport, storage and disposal10.1 Transport Use the original

Transport, storage and disposalEppendorf BioSpectrometer® fluorescenceEnglish (EN)82

83Technical dataEppendorf BioSpectrometer® fluorescenceEnglish (EN)11 Technical data11.1 Power supply11.2 Ambient conditionsDo not expose to direct su

Technical dataEppendorf BioSpectrometer® fluorescenceEnglish (EN)8411.4 Photometric properties11.5 FluorimeterMeasuring principle Single beam absorpti

85Technical dataEppendorf BioSpectrometer® fluorescenceEnglish (EN)11.6 Further technical parametersCuvette material For measurements in the UV:Quartz

Technical dataEppendorf BioSpectrometer® fluorescenceEnglish (EN)8611.7 Application parametersMethods Preprogrammed and freely programmable methods fo

87Evaluation procedureEppendorf BioSpectrometer® fluorescenceEnglish (EN)12 Evaluation procedureThis chapter describes the evaluation procedures avail

Evaluation procedureEppendorf BioSpectrometer® fluorescenceEnglish (EN)8812.1.3 Cuvette correctionAll absorbance values which are used for result calc

89Evaluation procedureEppendorf BioSpectrometer® fluorescenceEnglish (EN)The factor is either entered directly as a parameter during the "Factor&

9Operating instructionsEppendorf BioSpectrometer® fluorescenceEnglish (EN)1.4 Abbreviations usedAAbsorbanceDNADeoxyribonucleic aciddsDNADouble-strande

Evaluation procedureEppendorf BioSpectrometer® fluorescenceEnglish (EN)9012.4 DilutionIn the measure samples method step. entered dilutions are consid

91Evaluation procedureEppendorf BioSpectrometer® fluorescenceEnglish (EN)The application of the evaluation procedure can be activated in the parameter

Evaluation procedureEppendorf BioSpectrometer® fluorescenceEnglish (EN)9212.5.3 Conversion to molar concentrations and nucleic acid quantitiesThe conv

93Evaluation procedureEppendorf BioSpectrometer® fluorescenceEnglish (EN)For ssDNA, RNA, Oligo:MM = calculated relative molar mass; unit: kDabp = ente

Evaluation procedureEppendorf BioSpectrometer® fluorescenceEnglish (EN)9412.6 Special evaluation procedures for the dye methods12.6.1 Calculating the

95Evaluation procedureEppendorf BioSpectrometer® fluorescenceEnglish (EN)12.6.3 Conversion to amounts of dyeThe quantity (dimensions) of dye in the en

Evaluation procedureEppendorf BioSpectrometer® fluorescenceEnglish (EN)9612.8 Fluorimetry12.8.1 RFU valuesRelative Fluorescence Unit: RFU values are a

97Ordering informationEppendorf BioSpectrometer® fluorescenceEnglish (EN)13 Ordering informationOrder no. (International)Order no. (North America)Des

Ordering informationEppendorf BioSpectrometer® fluorescenceEnglish (EN)98

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