Jenway Genova Nano German User Manual
Genova Plus
Spectrophotometer
Operating Manual
736 505 REV A/11-12
Safety
Please read this information carefully prior to installing or using this equipment.
1. The unit described in this manual is designed be operated only by trained personnel. Any
adjustments, maintenance and repair must be carried out as defined in this manual, by a person qualified
to be aware of the hazards involved.
2. It is essential that both operating and service personnel employ a safe system of work, in addition to
the detailed instructions specified in this manual.
3. Other than for those items defined in the maintenance procedures herein there are no user serviceable
items in this instrument. Removal of covers and attempted adjustment or service by unqualified personnel
will invalidate the warranty and may incur additional charges for repair.
4. References should always be made to the Health and Safety data supplied with any chemicals used.
Generally accepted laboratory procedures for safe handling of chemicals should be employed.
5. If it is suspected that safety protection has been impaired in any way, the unit must be made inoperative
and secured against any intended operation. The fault condition should immediately be reported to the
appropriate servicing authority.
Merci de lire attentivement ces informations avant d’installer ou d’utiliser cet appareil.
1. L’appareil décrit dans ce manuel est conçu pour être utilisé uniquement par des personnes formées. Tout
réglage, maintenance ou réparation doit être effectué comme décrit dans ce manuel, par une personne
qualifiée consciente des risques encourus.
2. Il est essentiel que les personnes utilisant et intervenant sur cet appareil respectent les règles de sécurité
de travail, en plus des instructions détaillées précisées dans ce manuel.
3. En-dehors des éléments décrits dans les procédures de maintenance ci-incluses, cet appareil ne contient
aucun élément réparable par l’utilisateur. L’enlèvement des capots et les tentatives de réglage ou de
réparation par des personnes non qualifiées invalide toute garantie et entraîne un risque de frais de
réparation supplémentaires.
4. Toujours se référer aux fiches techniques de santé et de sécurité accompagnant tout produit chimique
utilisé. Respecter les procédures de laboratoire généralement acceptées pour la manipulation en toute
sécurité des produits chimiques.
5. Si l’utilisateur suspecte qu’un problème quelconque puisse mettre en cause la sécurité, l’appareil doit
être rendu inopérant en empêchant son utilisation. Communiquer la défaillance constatée au service de
maintenance compétent.
Bitte lesen Sie diese Hinweise vor Installation oder Gebrauch dieser Ausrüstung sorgfältig durch.
1. Das in diesem Handbuch beschriebene Gerät darf nur von geschultem Personal bedient werden.
Alle Anpassungen, Wartungsarbeiten und Reparaturen müssen entsprechend der Vorgaben in diesem
Handbuch und von einer kompetenten Person, die mit den damit verbundenen Gefahren vertraut ist,
durchgeführt werden.
2. Es ist wichtig, dass sowohl das Bedienungs- als auch das Service-Personal zusätzlich zu den detaillierten
Anweisungen in diesem Handbuch ein sicheres Arbeitssystem einsetzen.
3. Mit Ausnahme der Teile, deren Wartungsverfahren in diesem Handbuch beschrieben sind, enthält dieses
Gerät keine weiteren Teile, die vom Benutzer gewartet werden können. Das Entfernen von Abdeckungen
und Versuche von hierfür unqualifiziertem Personal, Anpassungen oder Wartungsarbeiten durchzuführen,
haben zur Folge, dass die Garantie verfällt und können zusätzliche Reparaturkosten auslösen.
1
4. Es ist jederzeit auf die sicherheitsrelevanten Daten sämtlicher verwendeter Chemikalien Bezug zu
nehmen. Allgemein anerkannte Labormethoden zum sicheren Umgang mit Chemikalien sollten eingesetzt
werden.
5. Besteht der Verdacht, dass die Sicherheitsvorrichtungen in irgendeiner Weise beschädigt wurden, muss
das Gerät außer Betrieb genommen und gegen weiteren Gebrauch gesichert werden. Die Störung sollte
der zuständigen Serviceeinrichtung unverzüglich gemeldet werden.
Leggere attentamente queste istruzioni prima di installare o utilizzare il dispositivo.
1. L’unità descritta nel presente manuale è stata realizzata per essere utilizzata solo da personale che ha
ricevuto l’apposita formazione. Qualsiasi operazione di regolazione, manutenzione e riparazione deve
essere effettuata sulla base di quanto indicato nel presente manuale da personale qualificato consapevole
dei rischi connessi.
2. È fondamentale che il personale operativo e il personale addetto alla manutenzione utilizzino un sistema
di lavoro sicuro, oltre a seguire le istruzioni specificate nel presente manuale.
3. Oltre a quelli indicati nelle procedure di manutenzione, all’interno di questo dispositivo non sono presenti
altri elementi sui quali è possibile effettuare interventi. La rimozione delle protezioni e qualsiasi tentativo
di regolazione o di manutenzione posto in essere da personale non qualificato invaliderà la garanzia. In
questi casi, sarà necessario pagare un importo per le riparazioni effettuate.
4. È sempre necessario fare riferimento ai dati sulla salute e sulla sicurezza forniti con le sostanze chimiche
utilizzate. Adottare le procedure di laboratorio generalmente accettate per la gestione delle sostanze
chimiche.
5. Nel caso in cui si sospetti che la salute possa essere pregiudicata in qualsiasi modo, disattivare l’unità per
renderla inutilizzabile. Qualsiasi condizione di errore deve essere immediatamente segnalata al responsabile
per la manutenzione.
Lea esta información atentamente antes de instalar o utilizar este equipo.
1. La unidad descrita en este manual está diseñada para que solamente la utilice personal con formación.
Cualquier operación de ajuste, mantenimiento y reparación debe llevarse a cabo del modo indicado en
este manual y debe realizarla una persona cualificada que sea consciente de los peligros que implica.
2. Es fundamental que tanto los operarios como el personal de servicio utilicen un sistema de trabajo
seguro, así como las instrucciones detalladas que se especifican en este manual.
3. Cualquier elemento que no se encuentre entre los definidos en los procedimientos de mantenimiento
aquí descritos no podrá utilizarse en este instrumento. La extracción de las tapas y los intentos de ajuste
o reparación por parte de personal no cualificado invalidarán la garantía y pueden incurrir en cargos
adicionales por reparación.
4. Siempre deberían consultarse los datos sobre Salud y Seguridad que se suministran con cualquier
producto químico que se utilice. Es necesario llevar a cabo los procedimientos de laboratorio de aceptación
generalizada para la manipulación segura de productos químicos.
5. Si existe la sospecha de que las medidas protectoras de seguridad han quedado dañadas en cualquier
modo, la unidad debe inutilizarse y protegerse contra toda operación que se intente llevar a cabo. El estado
de fallo debe comunicarse inmediatamente a la autoridad de servicio de mantenimiento y reparación
pertinente.
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Contents
Page
Safety 1
SECTION 1 - Introduction 8
1.1 INSTRUMENT DESCRIPTION 8
1.2 INSTRUMENT SPECIFICATION 8
SECTION 2 – Installation 10
2.1 UNPACKING 10
2.2 INSTALLATION 10
2.3 DISPLAY 11
2.4 CONTROLS 12
2.5 REAR PANEL 13
2.6 FRONT PANEL 13
SECTION 3 – Theory and Practice of Spectroscopy Measurements 14
3.1 THEORY OF SPECTROSCOPY MEASUREMENT 14
3.2 NUCLEIC ACID DETERMINATION 14
3.3 SPECTROSCOPY MEASUREMENT 15
3.4 GOOD PRACTICE GUIDELINES 16
SECTION 4 – Instrument Setup 18
4.1 NAVIGATING AND SCREEN SETUP 18
4.2 TIME AND DATE 19
4.3 INSTRUMENT SETTINGS MENU 20
4.4 SECURITY AND SETTING PASSWORDS 20
4.4.1 Setting Security Codes 20
4.4.2 Settings lock 20
4.4.3 Method Lock 21
4.5 MODE SELECTION 21
4.6 GLP SETTINGS 22
4.7 SCREEN CONTRAST 22
SPECTROPHOTOMETER MENU OPTIONS 23
SECTION 5 – PHOTOMETRICS 23
5.1 MODE SPECIFIC PARAMETERS 23
5.2 METHOD SET UP 23
5.2.1 Selecting a Wavelength 23
5.3 CALIBRATION 24
5.4 SAMPLE MEASURMENT 24
SECTION 6 – Concentration 25
6.1 MODE SPECIFIC PARAMETERS 25
6.2 METHOD SETUP 25
6.2.1 Selecting a Wavelength 25
6.2.2 Settings 26
6.2.2.1 Selecting Concentration Units 26
6.2.2.2 Changing the Resolution 26
6.2.2.3 Using a Standard 27
6.2.2.4 Using a Factor 27
6.3 CALIBRATION 27
6.3.1 Calibrating to a Standard 28
6.3.2 Calibrating to a Factor 28
3
6.4 SAMPLE MEASUREMENT 28
6.4.1 Measuring a Sample After Calibrating to a Standard 28
6.4.2 Measuring a Sample After Calibrating to a Factor 29
SECTION 7 – Spectrum 30
7.1 MODE SPECIFIC PARAMETERS 30
7.2 METHOD SETUP 30
7.2.1 Scan Settings 31
7.2.1.1 Selecting Absorbance or % Transmittance 31
7.2.1.2 Setting Start and End Wavelengths 31
7.2.1.3 Setting the Scan Interval 32
7.2.1.4 Y-Axis Scaling 32
7.3 CALIBRATION 33
7.4 SAMPLE MEASUREMENT 33
7.5 DATA ANALYSIS 34
7.5.1 Peaks and Valleys Threshold 34
7.5.2 Peaks and Valleys Table 34
7.5.3 Spectral Points Analysis 35
SECTION 8 – Quantitation 37
8.1 MODE SPECIFIC PARAMETERS 37
8.2 METHOD SETUP 37
8.2.1 Quantitation Settings 38
8.2.1.1 Selecting Absorbance or % Transmittance 38
8.2.1.2 Selecting a Wavelength 38
8.2.1.3 Selecting Concentration Units 38
8.2.1.4 Changing the Resolution 38
8.2.1.5 Selecting the Number of Replicate Standard Measurements 39
8.2.1.6 Selecting Automatic or Manual Replicate Measurements 39
8.2.1.7 Selecting Number of Standards 39
8.2.2 Quantitation Table 39
8.2.2.1 Editing Standard Data 39
8.2.2.2 Creating a New Standard Curve 40
8.2.3 Standard Curve 41
8.3 CALIBRATION 42
8.4 SAMPLE MEASUREMENT 42
8.5 DATA ANALYSIS 43
SECTION 9 – Kinetics 44
9.1 MODE SPECIFIC PARAMETERS 44
9.2 METHOD SET UP 44
9.2.1 Kinetics Settings 45
9.2.1.1 Y-Axis Scaling 45
9.2.1.2 Setting Lag Time or Start on Level 46
9.2.1.3 Selecting Absorbance or % Transmittance 46
9.2.1.4 Changing the Resolution 46
9.2.1.5 Selecting Concentration Units 47
9.2.1.6 Using a Standard 47
9.2.1.7 Using a Factor 47
9.2.1.8 Selecting a Wavelength 48
9.2.1.9 Setting the Kinetics Measurement Time 48
9.3 CALIBRATION 48
9.4 SAMPLE MEASUREMENT 48
9.5 DATA ANALYSIS 49
4
SECTION 10 – MULTI-WAVELENGTH 51
10.1 MODE SPECIFIC PARAMETERS 51
10.2 METHOD SET UP 51
10.2.1 Multi-Wavelength Settings 52
10.2.1.1 Setting the Number of Wavelengths 52
10.2.1.2 Setting the Measurement Wavelengths 52
10.2.1.3 Changing the Resolution 52
10.2.1.4 Selecting Concentration Units 52
10.2.1.5 Setting the Concentration Calculation Equation and Factors 53
10.3 CALIBRATION 53
10.4 SAMPLE MEASUREMENT 53
LIFE SCIENCE MENU OPTIONS 54
SECTION 11 – Concentration Plus 54
11.1 MODE SPECIFIC PARAMETERS 54
11.2 METHOD SETUP 54
11.2.1 Selecting a Wavelength – Operating Menu 54
11.2.2 Concentration Plus Settings 55
11.2.2.1 Selecting a Wavelength – Concentration Plus Settings 55
11.2.2.2 Selecting Concentration Units 55
11.2.2.3 Changing the Resolution 56
11.2.2.4 Using a Standard 56
11.2.2.5 Using a Factor 56
11.2.2.6 Setting the Dilution Factor 57
11.3 CALIBRATION 57
11.3.1 Calibrating to a Standard 57
11.3.2 Calibrating to a Factor 58
11.4 SAMPLE MEASUREMENT 58
11.4.1 Measuring a Sample After Calibrating to a Standard 58
11.4.2 Measuring a Sample After Calibrating to a Factor 59
SECTION 12 – PURITY SCAN 60
SECTION 13 – MULTI-WAVELENGTH PLUS 61
13.1 MODE SPECIFIC PARAMETERS 61
13.2 METHOD SET UP 61
13.2.1 Multi-wavelength Plus Settings 62
13.2.1.1 Setting the Measurement Wavelengths 62
13.2.1.2 Changing the Resolution 62
13.2.1.3 Selecting Concentration Units 62
13.2.1.4 Setting the Dilution Factor 63
13.2.1.5 Setting the Reference Wavelength 63
13.2.1.6 Setting the Concentration Calculation Equation and Factors 63
13.3 CALIBRATION 64
13.4 SAMPLE MEASUREMENT 64
SECTION 14 – DNA 65
14.1 DNA MENU OPTIONS 65
14.2 dsDNA 65
14.3 ssDNA 65
14.4 RNA 66
14.5 OLIGONUCLEOTIDES 66
14.6 260 / 280 67
14.7 260 / 230 67
14.8 VARIABLE RATIO 67
14.9 CALIBRATION AND SAMPLE MEASUREMENT 68
5
SECTION 15 – PROTEIN 69
15.1 PROTEIN MENU OPTIONS 69
15.2 PIERCE 660 ASSAY 69
15.3 BCA ASSAY 69
15.4 BRADFORD ASSAY 70
15.5 LOWRY ASSAY 70
15.6 BIURET ASSAY 71
15.7 DIRECT UV 71
15.8 CALIBRATION AND SAMPLE MEASUREMENT 71
SECTION 16 – OD 600 72
16.1 MODE SPECIFIC PARAMETERS 72
16.2 METHOD SETUP 72
16.2.1 Selecting a Wavelength 72
16.2.2 Settings 73
16.2.2.1 Using a Standard 73
16.2.2.2 Using a Factor 74
16.2.2.3 Using an Instrument Factor 74
16.2.2.4 Setting the Dilution Factor 75
16.3 CALIBRATION 75
16.3.1 Calibrating to a Standard 76
16.3.2 Calibrating to a Factor 76
16.4 SAMPLE MEASUREMENT 76
16.4.1 Measuring a Sample After Calibrating to a Standard 76
16.4.2 Measuring a Sample After Calibrating to a Factor 77
SECTION 17 – SAVING, PRINTING AND AUTOLOGGING 78
17.1 SAVING METHODS 78
17.1.1 Saving Methods To Internal Memory 78
17.1.2 Saving Methods to USB Memory Stick 79
17.2 OPENING METHODS 80
17.2.1 Opening Methods From Internal Memory 80
17.2.2 Opening Methods From USB Memory Stick 80
17.3 DELETING METHODS 81
17.4 SAVING RESULTS 81
17.5 OPENING RESULTS 82
17.6 DELETING RESULTS 83
17.7 PRINTING 83
17.7.1 Print Setup 83
17.7.1.1 Print Setup – PHOTOMETRICS, CONCENTRATION, MULTIWAVELENGTH AND OD 600 84
17.7.1.2 Print Setup - SPECTRUM AND PURITY 84
17.7.1.3 Print Setup – QUANTITATION AND PROTEINS 84
17.7.1.4 Print Setup – KINETICS 84
17.7.2 Printing Results 85
17.8 AUTOLOGGING 85
17.8.1 Setting the Number of Sample Repetitions 85
17.8.2 Selecting Result’s Destination 87
17.9 LOCKED METHODS 87
17.10 CONNECTING TO A PC 87
SECTION 18 – ACCESSORIES AND SPARE PARTS 88
18.1 OPTIONAL ACCESSORIES 88
18.2 CONNECTING THE ACCESSORIES 88
18.2.1 Internal Printer 88
18.2.2 Passive Accessories 89
18.2.3 Active Accessories 89
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18.2.3.1 Automatic 8 cell turret 90
18.2.3.2 Peltier 90
18.2.3.3 Sipper pump 91
18.2.3.4 Combined sipper Peltier pump 93
18.3 USING THE ACCESSORIES 93
18.3.1 Automatic 8 cell turret 93
18.3.1.1 Automatic 8 cell turret – supporting creation of a standard curve in quantitation 94
18.3.2 Peltier 94
18.3.3 Sipper pump 95
18.3.3.1 Manual Sipper Pump Settings 95
18.3.3.2 Timed Sipper Pump Settings 96
18.3.4 Combined sipper Peltier 98
18.4 SPARES 99
SECTION 19 – MAINTENANCE AND SERVICE 100
19.1 ROUTINE MAINTENANCE 100
19.2 LAMP REPLACEMENT 100
19.2.1 Xenon Lamp Module Replacement 100
19.3 FIRMWARE UPDATE PROCEDURE 100
19.4 SERVICE 100
SECTION 20 – TROUBLESHOOTING 101
20.1 ERROR CODES 101
20.2 TROUBLESHOOTING GUIDE 103
20.3 TECHNICAL SUPPORT 103
SECTION 21 – DECLARATION OF CONFORMITY 104
SECTION 22 – GLOSSARY OF ICONS 105
7
SECTION 1 - Introduction
INSTRUMENT DESCRIPTION1.1
The Genova Plus is a UV/visible spectrophotometer dedicated to life science analysis. This
spectrophotometer allows the measurement of DNA concentrations and purity ratios using
wavelengths recorded at 260, 280 and 230nm, with an optional correction at 320nm.The Genova Plus
is pre-programmed with Bradford, Lowry, Biuret, BCA and Direct UV methods for protein analysis. The
Genova Plus has an OD measurement mode enabling users to measure optical density at 600nm for
cell harvesting. The purity scan across the entire wavelength range from 198 to 1000nm displays any
distorted peaks enabling impurities to be easily identified.
This life science spectrophotometer uses icon driven software and has an improved navigation
system for easy and intuitive usability. As well as the dedicated life science measurement modes
this instrument can also be used as a standard spectrophotometer with measurement modes for
photometrics, concentration, multi-wavelength, spectrum scanning, quantitation and kinetics.
INSTRUMENT SPECIFICATION1.2
Genova Plus
Wavelength
Range 198 to 1000nm
Resolution 1nm
Accuracy ± 2nm
Repeatability ± 0.5nm
Spectral bandwidth 5nm
Photometrics
Transmittance 0 to 199.9%
Absorbance -0.300 to 2.500A
Accuracy* ±1%T, ±0.01Abs at 1.000 Absorbance
Resolution 0.1%T, 0.001A
Stray light* <0.5% at 340nm and 220nm
Concentration/Concentration Plus
Range 0 to 9999
Resolution Selectable 1/0.1/0.01/0.001
Calibration Blank with a single standard or factor
Units no units, %, ppm, EBC, SRM, mEq/l, mEq, M, mM,
µM, nM, U, U/l, U/ml, g/l, mg/l, µg/l, ng/l, g/dl,
mg/dl, µg/dl, mg/ml, µg/ml, ng/ml, µg/µl, ng/µl,
mol/l, mmol/l,
Factor 0.001 to 10000
Standard 0.001 to 1000
Quantitation
Range 0 to 9999
Resolution Selectable 1/0.1/0.01/0.001
Calibration Blank with up to 12 standards
Units no units, %, ppm, EBC, SRM, mEq/l, mEq, M, mM,
µM, nM, U, U/l, U/ml, g/l, mg/l, µg/l, ng/l, g/dl,
mg/dl, µg/dl, mg/ml, µg/ml, ng/ml, µg/µl, ng/µl,
mol/l, mmol/l
8
Curve fit algorithms Quadratic, quadratic through zero, linear, linear
through zero, interpolate
Multi-wavelength
Range 0 to 9999
Data points Up to 4 wavelengths
Calculations Sum, product, ratio, difference
Kinetics
Measurement Time 2 to 9999 seconds
Calibration Blank with a single standard or factor
Display Graphical and concentration
Analysis Concentration, rate of change, initial and final
absorbance/%T
Resolution Selectable 1/0.1/0.01/0.001
Spectrum/Purity Scan
Range 198 to 1000nm
Scan interval Selectable 1, 2 or 5nm
Analysis Absorbance or % transmittance and peak and valley
wavelengths
Multi-wavelength Plus
Range 0 to 9999
Data points 3 wavelengths + optional reference wavelength
Calculations concentration, ratio
DNA
Measurement modes dsDNA, ssDNA, RNA, Oligonucleotides,
260/280, 260/230, Variable Ratio
Protein
Measurement modes Pierce 660, BCA, Bradford, Lowry, Biuret, Direct UV
OD 600
Range 0.00 E-19 to 9.99 E+19
Calibration Blank with a single standard or factor
Units cells/ml
Factor 0.01 E-19 to 9.99E+19
Standard 0.01 E-19 to 9.99E+19
Instrument Factor 0.001 to 9999.999
Other
Beam height 15mm
Light source Xenon lamp
GLP Current time and date, user ID, settings lock and
method lock
Number of users 999
Methods memory 312 (including pre-programmed methods)
Results memory Limited by attached mass storage device
Removable media USB (supplied)
Outputs USB, Analogue, RS232, Internal printer
Power 24V
Size (w x d x h) 275 x 400 x 220mm
Weight 6kg
*Assessment must be performed with a 630 204 - 10 x 10mm path length cuvette holder installed.
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SECTION 2 – Installation
2.1 UNPACKING
Remove the Genova Plus from the packaging and ensure the following items are included:
1. Model Genova Plus spectrophotometer fitted with micro-cuvette holder (736 501)
2. 24V 65W power supply unit (021 060)
3. Pack of 100 disposable UV micro-volume cuvettes 70µl (035 143)
4. 4GB USB memory stick (019 146)
5. Instruction manual (736 505)
6. Jenway Foreign Manual CD (JENMANCD)
7. 10x10mm single cuvette holder (630 204)
2.2 INSTALLATION
The Genova Plus is supplied ready to use.
The unit should be placed on a clean flat surface which is free from drafts and vibrations. The units are
designed for operation on 90V to 264V AC input at 47 to 63Hz. Select the correct plug attachment and
attach to the power supply unit as shown below:
10
Fig 2.2.1 – Power supply unit with various plugs
Connect the power supply unit to the power inlet socket on the rear panel of the instrument and
connect to the mains socket. Turn the power on at the mains and switch the instrument on using the
power switch on the rear of the instrument.
The instrument will initially check for firmware updates (Section 20.3) and then perform several poweron tests before displaying the main menu:
1. Instrument check – ensures the validity of the saved parameters
2. Dark test
3. Checks for the accessory fitted. If an active accessory is found the instrument verifies communication
and response
4. Self calibration of wavelengths
5. Checks communication between USB memory stick port and the instrument
2.3 DISPLAY
1
2
5
Fig 2.2.2 – All Power On Tests Complete
3
4
The instrument has a dot matrix display which enables icons and graphs to be displayed clearly. Following
successful completion of the power on tests the main menu screen will be displayed:
Fig. 2.3.1 – Display
11
Life Science/Spectrophotometer menu options
1. Purity/Spectrum measurement mode
2. Concentration plus/Concentration measurement mode
3. Back key
4. Time and date toggle and settings
5. Multi-wavelength plus/Multi-wavelength measurement mode
6. Toggle between Life Science and Spectrophotometer modes
7. Protein/Photometrics measurement mode
8. Instrument settings menu
9. DNA/Quantitation measurement mode
10. OD 600/Kinetics measurement mode
2.4 CONTROLS
The keypad used for this model enables an easy and effective way of navigating the different measurement
modes, entering numbers, saving and analysing results. The soft keys are active when an icon is displayed
above or adjacent to the key. The only exception to this is the back key which is always active.
The main Life Science menu screen and surrounding keypad is displayed below.
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Fig. 2.4.1 – Display
Life Science/Spectrophotometer menu options
1. Purity/Spectrum measurement mode
2. Concentration plus/Concentration measurement mode
3. Back key
4. Time and date toggle and settings
5. Multi-wavelength plus/Multi-wavelength measurement mode
6. Toggle between Life Science and Spectrophotometer modes
7. Protein/Photometrics measurement mode
8. Instrument settings menu
9. DNA/Quantitation measurement mode
10. OD 600/Kinetics measurement mode
2.5 REAR PANEL
The image below shows the rear panel on the instrument:
Fig. 2.5.1 – Rear Panel
1. Lamp access panel Allows access to lamp when replacement is necessary
2. Power switch On/off switch for the unit
3. Power in socket Connection socket for power supply unit
4. RS232 serial port Connection to a PC or external serial printer
5. Output sockets Analogue output
2.6 FRONT PANEL
The image below shows the front panel of the instrument:
1
2
3
5
4
Fig. 2.6.1 – Front Panel
1. Integral printer (optional accessory)
2. Keypad
3. USB memory stick slot
4. Instrument lid
5. Display
13
SECTION 3 – Theory and Practice of Spectroscopy
Measurements
3.1 THEORY OF SPECTROSCOPY MEASUREMENT
UV-visible spectroscopy is the measurement of the absorbance of light at a specific wavelength in a
sample. This is used to identify the presence and concentration of molecular entities within the sample.
The Beer-Lambert law is used to relate the absorption of light to the properties of the sample through
which the light is travelling through. The Beer-Lambert law states that:
A is the absorbance
is the molar absorption coefficient (l mol-1cm-1)
c is the concentration (mol l-1)
l is the path length (cm)
This law shows that absorbance is linear to concentration but this is only true for low concentrations. For
absorbance levels above 3 the concentration starts to move away from the linear relationship.
Transmittance is the proportion of the light which passes through the sample:
I
o
L
Therefore: T = I
Io
Absorbance is inversely related to transmittance:
A = log 1
T
3.2 NUCLEIC ACID DETERMINATION
DNA, RNA and oligonucleotides can be measured directly in aqueous solutions in a diluted or undiluted
form. Aqueous buffers with low ion concentrations (e.g. TE buffer) are ideal for this method. The
concentration is commonly determined by measuring at 260nm against a blank and then evaluating
against a factor.
t
I
t
Where:
Io is the incident light
lt is the transmitted light
L is the path length
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The Genova Plus has pre-defined methods installed which assume that absorption of 1 OD (A) is equivalent
to approximately: 50µg/ml dsDNA, 37µg/ml ssDNA, 40µg/ml RNA and 30µg/ml for oligonucleotides.
DNA interference by contaminants can be assessed by the calculation of an absorption ratio. The ratios
A260/A280 and A260/A230 are used to estimate the purity of nucleic acids, since proteins absorb at
280nm and substances such as peptides, phenols, aromatic compounds or carbohydrates absorb at
230nm. Pure DNA should have an A260/A280 ratio of approximately 1.8 and pure RNA 2.0. In pure
nucleic acid samples the A260/A230 ratio should be approximately 2.2.
Nucleic acid concentration can also be estimated with the following calculations:
Conc (µg/ml) = (Abs@260nm x 62.9) - (Abs@280nm x 36.0)
Conc (µg/ml) = (Abs@260nm x 49.1) - (Abs@230nm x 3.48)
Referring to a blank value where no absorption should occur is commonly required. On the Genova Plus
the default reference wavelength is 320nm and the user can include the measured absorbance value in
all nucleic acid calculations. The default wavelength can be modified from 320nm if required.
3.3 SPECTROSCOPY MEASUREMENT
There are four main components of a spectrophotometer. These are a light source to emit a high and
constant amount of energy over the full wavelength range; a method for separating the light into
discreet wavelengths; a sample holder and a light detector.
The optical layout of the Genova Plus spectrophotometer is shown:
Figure 3.3.1 – Diagram of light path
The light from the pre-aligned xenon lamp is focused onto the grating, with 1200 lines per millimeter,
which separates the light into discrete wavelengths. The diffracted spectrum of light then passes through
a further slit and lens arrangement before passing through the sample in the sample chamber from left
to right. The light which is not absorbed by the sample is transmitted through a collecting lens and onto
the signal detector. The photo-diode detector used is mounted directly onto the detector PCB and the
output is used to calculate the % transmittance. The result is displayed either as % transmittance or
absorbance on the instrument display.
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3.4 GOOD PRACTICE GUIDELINES
1. For optimum performance all spectrophotometers should be sited in a clean, dry, dust free
atmosphere. When in use ambient temperature and light levels should remain as constant as possible.
2. If required adherence to Standard Operating Procedures (SOP) and Good Laboratory Practice (GLP)
should be monitored with regular calibration checks and a suitable Quality Control (QC) programme.
3. The sample chamber lid must be fully closed during measurement and before any readings are recorded
or printed.
4. The correct selection of sample containers is imperative for accurate and reproducible results:
a) Check that the material of the sample container is compatible with the wavelengths to be used
for measurement. In general glass can only be used down to 360nm or 320nm depending on quality.
Standard plastic cuvettes can be used down to 320nm. Special UV versions can be used down to 260nm.
Below this level quartz cuvettes must be used.
b) Plastic disposable cuvettes should only be used ONCE.
c) Glass cuvettes should be thoroughly cleaned after use. Discard when scratches become evident on
optical surfaces.
d) Care should be taken when selecting semi-micro or micro cuvettes. The cuvette window on the inner
chamber (the area filled with sample) must be wider than the aperture in the sample holder or light will
reach the detector without passing through the sample. In this case, semi-micro or micro cuvettes with
self-screening black surrounds must be used or, alternative holders for these cuvettes should be used.
e) Glass test tubes and other sample tubes should be used with care. Where possible, matched tubes
should be used and any index mark set to the correct position before measurements are made.
f) Ensure any sample containers used are compatible with the constituents of both the samples and
standards they are to hold. Plastic cuvettes are not compatible with organic solvents.
g) All sample containers must be handled with care; by the top, bottom and non-optical surfaces only.
Any finger marks evident must be removed by a suitable cleaning process.
h) Flow-through cuvettes must be selected with care and consideration for the sample type, sample
volume, pumping system, rinse, sample and waste handling to be used.
5. Samples and standards should not be stored in open cuvettes or sample containers as evaporation will
change the value and lead to staining of the walls which may be irreversible. If stored in stoppered and
sealed cuvettes, they should be filled with little or no air space and the values regularly checked against
a reference standard or quality control material.
6. Samples should be allowed to equilibrate to ambient temperature before measurement (unless a
suitable temperature controlled sample holder is in use). Temperature change during measurement may
cause air bubbles to form on the walls of the sample holder. This is a common cause of drift during
measurement.
7. In the preparation of samples and standards high grade borosilicate glass and AR grade chemicals
and reagents must be used. Good quality deionised water or other suitable solvents must be used for
dissolving or diluting samples, chemicals and reagents.
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8. All measurements require calibration to a blank, for maximum accuracy this should be prepared with
care using the same deionised water or solvent used for dissolving or diluting the sample. Where reagents
are added to the sample to produce a colour proportional to its concentration a ‘sample based’ blank
should be used. In this case the blank should consist of all reagents or chemicals to be used, except the
sample which will produce the colour to be measured.
9. Deviations from the Beer-Lambert Law may occur at high and low concentrations giving non-linear
response during sample concentration measurements. For all new methods a linear range should be
defined by the preparation of a calibration curve. The quantitation mode may be used to construct such
a curve against which sample results are automatically measured.
10. Cuvettes and sample holders must be filled to a minimum level which covers the light path. All
Jenway spectrophotometers have a beam height of 15mm.
11. The instrument must be calibrated to zero absorbance/100% transmittance prior to taking readings.
In the spectrum measurement mode a baseline scan must be performed before performing a sample
scan.
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SECTION 4 – Instrument Setup
4.1 NAVIGATING AND SCREEN SETUP
The main life science menu screen is displayed below.
Fig 4.1.1 – Life Science Home Screen
Fig 4.1.2 – Spectrophotometer Home Screen
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To navigate around the spectrophotometer screen press the soft keys adjacent to icons displayed on
the screen. There is a back key which returns to the previous menu without saving any changes.
The main menu screens provide access to all measurement modes, the time and date menu and the
instrument settings menu. The measurement modes are specific to each of the instrument’s two home
screens. The life science home screen gives access to the purity scan, concentration plus, multi-wavelength
plus, protein, DNA and OD 600 modes, whereas the spectrophotometer home screen gives access to
the spectrum, photometrics, quantitation, concentration, multi-wavelength and kinetics modes. The
instrument settings menu enables access to settings lock, security codes, method lock, mode selection,
user ID and screen contrast menus.
When a measurement mode is opened the operating
menu enables changes to measurement parameters
and settings to be made. Depending on the mode, the
measurement parameters can be accessed through the
settings menu which is displayed in the top right hand
corner of the screen. The only mode where this function
is not available is the photometrics mode; instead a toggle
Operating Menu
(Photometrics measurement mode)
icon is displayed which is used to change the primary and
secondary displays. The DNA and protein modes require
the user to initially select a method before the operating
menu option is available.
The utility toolbar is displayed on the left hand side of the operating menu and provides the same
functions in all of the measurement modes. This toolbar enables access to printing, print setup options,
opening, saving and deleting results and methods and autologging options. For more details on the
different functions of the utility toolbar refer to section 17.
4.2 TIME AND DATE
The time and date menu enables the current time and date
to be set. This information will be saved on all results and
displayed on printouts. The time and date menu can be
accessed from the main menu by holding the key below
the time and date icon for 2 seconds. Pressing the key
once cycles the display between time and date.
In the time and date menu to set the time press the key
adjacent to the clock icon. Select the digit to be changed
using the keys at the bottom of the screen. Use the keys
adjacent to the arrow icons to increase or decrease the
number. The clock function uses a 24 hour format.
In the time and date menu to set the date press the
key adjacent to the calendar icon. Select the digit to be
changed using the keys at the bottom of the screen.
Use the keys adjacent to the arrow icons to increase or
decrease the number. The date format can be displayed as
either European dd/mm/yy or American mm/dd/yy.
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To change between the two formats press the key below the toggle icon. Once the current time and
date have been set press the key adjacent to the tick icon to save the changes. To exit this menu without
saving any changes press the back key and the screen will return to the main menu.
4.3 INSTRUMENT SETTINGS MENU
The instrument settings menu is accessed by pressing the key below the instrument settings icon in the
main menu. This menu enables access to settings lock, security code, method lock, mode selection, user
ID and screen contrast menus. The tick icon saves any changes made and returns to the main menu.
Settings lock
Security code
Method lock
Mode selection
Fig 4.3.1 - Settings Menu
4.4 SECURITY AND SETTING PASSWORDS
4.4.1 Setting Security Codes
Tick icon
User ID
Screen contrast
The security code function enables a security code to
be set to lock the instrument settings and measurement
mode settings. The security code is not specific to the user
ID but is designed to enable an administrator to control
either the instrument or protocols. The security code menu
is accessed through the instrument settings menu.
4.4.2 Settings lock
The settings lock function enables the instrument and measurement mode settings to be locked to
prevent any changes to the measurement parameters or instrument settings. The only exceptions to this
are that the user ID and contrast can be changed when the settings lock is active.
In the instrument settings menu press the key adjacent
to the security code icon. Using the keys at the bottom
of the screen select the digit to be changed. Use the keys
adjacent to the arrow icons to increase or decrease the
selected number. Once the preferred code has been set
press the key adjacent to the tick icon to save the security
code.
The settings lock function is accessed through the
instrument settings menu by pressing the key adjacent to
the open padlock icon. One press will lock the settings
instantly. To unlock the settings press the key again. This
will open the security code menu as detailed in section
4.4.1. The previously set security code must be entered
to unlock the settings. When the settings lock is active
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methods can still be opened, deleted and saved but the method parameters cannot be changed.
If the settings are locked before the security code has been set a default code of 660 will unlock the
settings.
4.4.3 Method Lock
key adjacent to the method lock icon again. The methods are now unlocked. If the settings lock is active
this must be disabled before the method lock can be activated or deactivated.
To enter the security code, use the keys at the bottom of
the screen to select the digit to be changed. Use the keys
adjacent to the arrow icons to increase or decrease the
selected number. Once the correct security code has been
entered press the key adjacent to the tick icon. The settings
are now unlocked.
When the method lock is active the method selection
menu is disabled in all the measurement modes therefore
methods cannot be opened, deleted or saved. However the
measurement parameters of the currently loaded method
can be changed. The method lock function is accessed
through the instrument settings menu by pressing the
key adjacent to the method lock icon. One press will lock
the methods instantly. To unlock the methods press the
In all the measurement modes if a user tries to save changes to a method when the method lock is active
the padlock icon flashes on the screen and changes cannot be saved.
4.5 MODE SELECTION
The mode selection function enables access to the various
measurement modes to be restricted. The required modes
can be selected and the settings lock activated to prevent
other users from accessing the deactivated modes. The
mode selection function can be accessed through the
instrument settings menu by pressing the key adjacent to
the mode selection icon. The measurement mode icons
which are displayed on the main menu are identified with a
mode shown icon. The mode icons which are not displayed
on the main menu are identified with a mode not shown
icon. To change a mode from displayed to restricted or vice
versa press the key adjacent to the measurement mode
icon. Once the required modes have been selected press
the key adjacent to the tick icon to save the changes. The
selected measurement modes will be displayed on the
main menu.
4.6 GLP SETTINGS
The same procedure can be used to restrict the mode
access in the spectrophotometer home screen.
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4.6 GLP SETTINGS
In addition to the time and date settings this instrument also has a user ID function. This function enables
an individual three digit ID number to be set. This will be displayed on all printouts and saved results.
4.7 SCREEN CONTRAST
The user ID function can be accessed through the
instrument settings menu by pressing the key adjacent to
the user ID icon. Use the keys at the bottom of the screen
to select the digit to be changed. Use the keys adjacent to
the arrow icons to increase or decrease the number. Once
the preferred user ID has been set press the key adjacent to
the tick icon to save and return to the instrument settings
menu.
The screen contrast function enables the brightness of the
screen to be set. In the instrument settings menu press
the key adjacent to the screen contrast icon. Use the
keys below the arrow icons to increase or decrease the
screen contrast. Once the required contrast level has been
reached press the key adjacent to the tick icon to save and
return to the instrument settings menu.
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SPECTROPHOTOMETER MENU OPTIONS
SECTION 5 – Photometrics
The photometrics measurement mode enables simple measurements of absorbance and % transmittance
to be performed. The sample is measured at one wavelength and at one point in time. There are no post
measurement calculations available in this measurement mode.
5.1 MODE SPECIFIC PARAMETERS
Operating Menu
The photometrics operating menu enables measurement
parameters to be changed. The utility toolbar on the left
hand side of the screen enables access to printing, print
setup options, results, methods and autologging options.
For more details on the different functions of the utility
toolbar refer to section 17.
Print/print settings
Results selection menu
Method selection menu
Autolog menu
5.2 METHOD SET UP
Calibrate
to zero
Fig 5.1.1 - Operating Menu
This measurement mode is very simple and the only
parameters which can be adjusted are the wavelength and
the display format.
The toggle icon enables the large primary display to be set
to show the absorbance or % transmittance.
To change the primary and secondary displays press the
key adjacent to the toggle icon. Repeat presses will cycle
the display between absorbance and % transmittance.
Measure
sample
Toggle
Increase wavelength
Decrease wavelength
5.2.1 Selecting a Wavelength
The wavelength can be adjusted by using the keys adjacent to the arrow icons to increase or decrease
the wavelength. Once the required wavelength has been selected a calibration can be performed.
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5.3 CALIBRATION
The calibration must be performed at the same wavelength
at which the sample will be measured. Insert a cuvette
containing the blank solution into the sample chamber and
close the instrument lid. Press the key below the calibrate
to zero absorbance icon. This sets the instrument to zero
absorbance and 100% transmittance.
Once the calibration is complete the measure sample icon appears and the sample can be measured. If the
wavelength is adjusted before a sample is measured the measure sample icon will disappear and the instrument
must be calibrated again at the new wavelength.
5.4 SAMPLE MEASURMENT
It is not possible to measure a sample before the instrument
has been calibrated at the selected wavelength. Once the
calibration has been performed the measure sample icon
is displayed and a sample can be measured. Remove the
cuvette containing the blank solution and place a cuvette
containing the sample to be measured in the sample
holder. Close the instrument lid and press the key below the
measure sample icon. Once the measurement is complete
the photometric result will be shown on the screen.
Subsequent samples can be measured in the same way. If the wavelength is adjusted between sample
measurements then the instrument must be calibrated again before more samples can be measured.
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SECTION 6 – Concentration
The concentration measurement mode enables simple measurements of absorbance and concentration
to be performed. In this measurement mode it is possible to calibrate against a standard of a known
concentration or use a known factor. The sample is measured at one wavelength at one point in time.
There are no post measurement calculations available in this measurement mode.
6.1 MODE SPECIFIC PARAMETERS
Operating Menu
The concentration operating menu enables measurement
parameters to be changed. The utility toolbar on the left
hand side of the screen enables access to printing, print
setup options, results, methods and autologging options.
For more details on the different functions of the utility
toolbar refer to section 17. The settings icon enables the
wavelength, units, resolution, standard or factor to be
set.
Print/print settings
Results selection menu
Method selection menu
Autolog menu
6.2 METHOD SETUP
6.2.1 Selecting a Wavelength
Calibrate to zero
or standard
Fig 6.1.1 - Operating Menu
The wavelength can be adjusted in the operating menu
or in the settings menu. To adjust the wavelength, in the
operating menu, use the keys adjacent to the arrow icons
to increase or decrease the wavelength.
Measure to
factor
Settings
Increase wavelength
Decrease wavelength
The settings menu is accessed through the operating menu
by pressing the key adjacent to the settings icon. In the
settings menu press the key below the wavelength icon.
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6.2.2 Settings
The settings menu enables the wavelength, units, resolution, standard or factor to be set and is accessed
from the operating menu by pressing the key adjacent to the settings icon. Once all of the required
settings have been entered press the key adjacent to the tick icon to save and return to the operating
menu.
This will open a number entry screen. Use the keys at the
bottom of the screen to select the digit to be adjusted.
Use the keys adjacent to the arrow icons to increase or
decrease the wavelength to the required number. Press
the key adjacent to the tick icon to save the changes and
return to the settings menu.
Tick icon
Standard menu
Selecting
concentration
units
Selecting
resolution
Fig 6.2.2.1 – Settings Menu
When setting the method parameters either the standard or the factor should be selected. The standard
should be used if the factor is not known as selecting this option will calculate the factor. If the factor is
known it is not necessary to measure a known standard’s concentration. When the standard or factor is
not selected the value should be set to 1.00.
6.2.2.1 Selecting Concentration Units
The units of concentration can be selected from a number of options: no units, %, ppm, EBC, SRM,
mEq/l, mEq, M, mM, µM, nM, U, U/l, U/ml, g/l, mg/l, µg/l, ng/l, g/dl, mg/dl, µg/dl, mg/ml, µg/ml, ng/ml,
µg/µl, ng/µl, mol/l, mmol/l.
In the settings menu, press the key below the units icon.
This opens the unit selection screen which displays all the
different units. Use the keys adjacent to the arrow icons
to navigate around the screen to select the required units.
Once the required units have been highlighted press the
key adjacent to the tick icon to save and return to the
settings menu. The selected unit will be displayed in the
operating menu along with absorbance and selected
wavelength.
Selecting
wavelength
Factor menu
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6.2.2.2 Changing the Resolution
The resolution that the concentration is displayed as can be selected from 1, 0.1, 0.01 or 0.001 by repeat
presses of the key below the resolution icon in the settings menu.
6.2.2.3 Using a Standard
For example 00 the first press of the key alters 10, the second press alters 01. Use the keys adjacent to
the arrow icons to increase or decrease the selected number. Standard values from 0.001 to 1000 can be
entered. The standard value can be reset to one by pressing the key adjacent to the 001 icon. Once the
standard value has been entered press the key adjacent to the tick icon to save and return to the settings
menu. The entered value is displayed in the settings menu adjacent to the standard icon.
A standard value should only be entered if the factor is not known. If the factor is known the standard
value should be set to 1.000.
6.2.2.4 Using a Factor
The standard menu enables the value of a standard to
be entered. This function is accessed by pressing the key
adjacent to the standard icon. This opens the extended
number entry screen. Use the keys at the bottom of the
screen to select the digit to be changed. The key below the
digits must be pressed twice to select the adjacent digit.
The factor menu enables a factor to be entered. This
function is accessed by pressing the key adjacent to the
factor icon. This opens the extended number entry screen.
Use the keys at the bottom of the screen to select the digit
to be changed. The key below the digits must be pressed
twice to select the adjacent digit.
For example 00 the first press of the key alters 10, the second press alters 01. Use the keys adjacent to
the arrow icons to increase or decrease the selected number. Factor values of 0.001 to 10,000 can be
entered. The factor value can be reset to one by pressing the key adjacent to the 001 icon. Once the
factor has been entered press the key adjacent to the tick icon to save and return to the settings menu.
The entered value is displayed in the settings menu adjacent to the factor icon.
If the factor is not known a standard should be measured in order to calculate the factor. If a standard
is used the factor value should be set to 1.000.
6.3 CALIBRATION
The calibration must be performed at the same wavelength at which the sample will be measured.
In the concentration measurement mode calibrations
against a standard or a factor can be performed following
a zero calibration. If the factor is not known calibration
against a known standard is performed in order to calculate
the factor. However if the factor is known there is no need
to calibrate using a standard.
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6.3.1 Calibrating to a Standard
Insert a cuvette containing the blank solution into the sample chamber and close the instrument lid. Press
the key below the calibrate to zero absorbance icon. The instrument will calibrate to zero absorbance.
Insert a cuvette containing the standard concentration sample solution into the sample chamber and
close the instrument lid.
Press the key below the calibrate to zero absorbance or
standard icon, this will open another menu with the option
to re-calibrate to zero absorbance or to calibrate to the
previously entered standard value. Press the key adjacent
to the calibrate to standard icon.
If the standard selected requires a factor beyond the
range of the instrument the check standard icon will be
displayed.
The instrument will take a reading and calibrate to the
standard concentration. Once the calibration is complete
the sample can be measured using the measure to standard
icon.
6.3.2 Calibrating to a Factor
Insert a cuvette containing the blank solution into the
sample chamber and close the instrument lid. Press the
key below the calibrate to zero absorbance icon. The
instrument will calibrate to zero absorbance. Once the
calibration is complete the sample can be measured using
the measure to factor icon.
6.4 SAMPLE MEASUREMENT
It is not possible to perform sample measurements before the instrument has been calibrated at the
selected wavelength. In this operating mode the type of sample measurement performed depends on
the calibration which has been carried out.
6.4.1 Measuring a Sample After Calibrating to a Standard
Remove the cuvette containing the standard sample and
place a cuvette containing the sample to be measured
in the sample chamber. Close the instrument lid and
press the key below the measure to standard icon. Once
the measurement is complete the concentration and
absorbance values are displayed.
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