Thermo Electron Evolution 300 User Manual Download Page 7

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Chapter 2 Spectrophotometer
Basics

This chapter describes the major components of your spectrophotometer.

Spectrophotometer

components

The following illustration identifies some major components visible on the
outside of a typical Evolution spectrophotometer. (Some components may
not be present on your spectrophotometer.)

Zero

Run

Keypad

LCD screen

Contrast

Optional printer

Floppy disk

drive

Sliding door

Sample

compartment

Port for optional

power supply

Thermo Electron Corporation

Evolution 300 and Evolution 600 User’s Guide

Summary of Contents for Evolution 300

Page 1: ......

Page 2: ...269 165900 ...

Page 3: ...12 Test tube vial holder 13 Rectangular cell holder 14 Fiber optic sampling accessory 14 Smart Accessories 18 System accessories 19 Installing Smart Accessories 20 Chapter 4 Using the Instrument 23 Local Control 23 Main Menu page 25 Text entry 33 Editing numeric fields 34 Pop up menus 35 File operations 35 Scan application 37 Fixed application 40 Quantitation application 43 Kinetics application 45...

Page 4: ...tenance 71 Routine maintenance 71 Cleaning instrument exterior 71 Internal printer 72 Light sources Evolution 600 only 73 Deuterium lamp 75 Tungsten halogen lamp 76 Mercury lamp 77 ii Evolution 300 and Evolution 600 User s Guide Thermo Electron Corporation ...

Page 5: ...on 300 or Evolution 600 spectrophotometer from Thermo Electron Our spectrophotometers integrate advanced hardware features with the power and flexibility of a wide range of Smart Accessories Thermo Electron Corporation Evolution 300 and Evolution 600 User s Guide 1 ...

Page 6: ...hardware or losing data Caution Indicates a potentially hazardous situation which if not avoided may result in minor or moderate injury It may also be used to alert against unsafe practices Warning Indicates a potentially hazardous situation which if not avoided could result in death or serious injury Danger Indicates an imminently hazardous situation which if not avoided will result in death or s...

Page 7: ...me major components visible on the outside of a typical Evolution spectrophotometer Some components may not be present on your spectrophotometer Ze ro Ru n 7 8 9 4 5 6 1 2 3 0 Keypad LCD screen Contrast Optional printer Floppy disk drive Sliding door Sample compartment Port for optional power supply Thermo Electron Corporation Evolution 300 and Evolution 600 User s Guide 3 ...

Page 8: ... connect a printer with a parallel interface Key Bd Use to connect an English keyboard this is supplied by Thermo Electron to meet electrical emission requirements Ext Use to connect an external TTL or contact closure trigger Accy Uses to connect an external accessory RS232 Use to connect a computer with an RS232 interface Ext Display Use to connect to a computer monitor 4 Evolution 300 and Evolut...

Page 9: ...nection Power cord Zer o Run 7 8 9 4 5 6 1 2 3 0 Danger Avoid shock hazard Always power off the spectrophotometer and disconnect the power supply from the wall outlet or power strip before disconnecting the power supply from the spectrophotometer Thermo Electron Corporation Evolution 300 and Evolution 600 User s Guide 5 ...

Page 10: ...show the locations of the connectors inside the sample compartment Evolution 300 Reference connector Sample connector Evolution 600 Sample connector Accessory detector connector Accessory detector connector Reference connector 6 Evolution 300 and Evolution 600 User s Guide Thermo Electron Corporation ...

Page 11: ...lled and removed as follows Manual accessories are mounted on a common baseplate This system ensures correct alignment of the accessory within the instrument automatically These accessories may be installed or removed without the need to switch off the instrument and may be installed in the sample front position and or the reference rear position The accessory is positioned by two pins A and B at ...

Page 12: ...e aligned over pins A and B in the instrument When properly aligned gently push the front end of the accessory downward until plate snaps into place under the toggle This positive location system ensures that the accessory is correctly aligned in the corresponding optical beam For removal of the accessory Grip the accessory through the finger hole in the accessory and pull it upwards For accessori...

Page 13: ...trument as described above 2 Access the white light function in the operating software Local Control From the Main Menu page select Utilities then press the White Light function key on the Utilities page VISION Select Command White Light The instrument will align the monochromator so that the zero order diffraction of the lamp s white light passes through the sample compartment This provides a bea...

Page 14: ...ning it counterclockwise moves the cell holder towards the rear When the cell holder is aligned properly the white light will not be seen on the edges of the cell holder 5 Tighten securing screw A to secure the cell holder position When you have finished aligning the accessory in the beam exit White Light mode by pressing White Light Off This returns the monochromator to its normal position ready ...

Page 15: ...rom 1 to 100mm The capacity of the cell holder is adjusted by turning the knob on the front which alters the space between the gripping blocks Turn the knob until the space is longer than the pathlength required insert the cell and reduce the space until the cell is held securely Important Do not overtighten Thermo Electron Corporation Evolution 300 and Evolution 600 User s Guide 11 ...

Page 16: ...designed to accommodate square cells with 8 8mm z dimension and 10mm pathlength and to operate at temperatures of 4 to 80 C 39 to 176 F Temperature control is achieved by means of an external temperature controlled circulator bath Refer to the Parts List supplied on CD with your instrument for details of circulators available from Thermo Electron A tubing kit for Evolution thermostatted accessorie...

Page 17: ...e Replace the sample compartment front panel ensuring that the tubes are correctly located in the slit s and are not being pinched between the front panel and the body of the instrument Ensure that the tubes do not obstruct the light beam If necessary secure the tubes using the self adhesive tubing clips supplied Test tube vial holder This accessory is designed for use with test tubes or vials fro...

Page 18: ...r allows the use of 15mm beam height filters and flowcells in the instrument Fiber optic sampling accessory This accessory enables samples to be measured outside of the sample compartment 14 Evolution 300 and Evolution 600 User s Guide Thermo Electron Corporation ...

Page 19: ...f the probe is manufactured to the same tolerances of a standard cuvette and are available in 5 10 and 20mm pathlengths VN Absolute Specular Reflectance accessory The VN Absolute Specular Reflectance accessory allows the absolute reflectance of samples as small as 12 5mm to be examined The VN accessory has the advantage of having light only strike the sample surface once and requires no calibrated...

Page 20: ...ontally outside the plane of the instrument A double beam Spectralon integrating sphere ensures high energy throughput and accuracy A transmittance port is also included for making diffuse transmittance measurements This DRA is fully CIE compliant for making color 16 Evolution 300 and Evolution 600 User s Guide Thermo Electron Corporation ...

Page 21: ...id samples in Transmittance mode A variety of sample holders are available allowing round square and odd shaped samples to be measured The industry standard 2 x 3 sample slides allow other accessories to be used with the Solid Sample Holder Thermo Electron Corporation Evolution 300 and Evolution 600 User s Guide 17 ...

Page 22: ... the drive motor control circuitry and memory There is also a socket for connecting a temperature probe accessory when purchased Smart Peltier Thermostatted Single Cell Holder For precise temperature control e g DNA denaturation this accessory provides temperature control from 5 to 110 C with ramping at 0 1 to 6 C per minute for a single 8 5mm cell and low head space micro cells There is also a so...

Page 23: ...ix fundamental mercury emission lines System accessories Some system accessories are installed by Thermo Electron Internal printer Located directly behind the LCD screen and is either factory or installed in the field by a qualified Thermo Electron service engineer Thermo Electron Corporation Evolution 300 and Evolution 600 User s Guide 19 ...

Page 24: ... the connector and socket during installation 1 Open the main lid of the sample compartment and remove any accessory that may be present in the sample front position 2 Remove the front wall of the sample compartment by sliding it upward in its grooves 3 Insert the tapered guide pin on the bottom front of the accessory into the corresponding guide pin hole located next to the D connector female Do ...

Page 25: ... Peltier Thermostatted Single Cell Holder the alignment holes and pins are not visible due to the plastic cover over the unit 5 Press down on the front of the unit to complete the connection between the accessory and the D connector 6 Replace the front wall of the sample compartment Thermo Electron Corporation Evolution 300 and Evolution 600 User s Guide 21 ...

Page 26: ...erature probe For removal of the accessory Grasp the handles of the accessory and pull it upwards The software will display a message confirming that the accessory has been removed Press Clear to remove this message 22 Evolution 300 and Evolution 600 User s Guide Thermo Electron Corporation ...

Page 27: ... to analyze samples The topics that are discussed include Local Control software VISIONsecurity and VISIONpro Local Control Function keys are operational in the context for each screen and are shown at the bottom of the display Thermo Electron Corporation Evolution 300 and Evolution 600 User s Guide 23 ...

Page 28: ...out additional prompts Zero Zero Perform the measurements specified in the current method Run Run Display the contents of the instrument library Library Clear error message or cancel pop up menus Clear Clear Confirm highlighted choices in menus and text entry boxes Enter Enter Select menu items and characters in text entry screens Arrow keys Enter numbers decimal point and minus sign Numeric keypa...

Page 29: ...or functions of the software are accessed from the Main Menu page Use the up and down arrow keys to highlight the required menu item and press Enter to confirm your selection The menu enables you to move between the six major applications and to view the contents of the Library or the contents of the disk currently in the drive Thermo Electron Corporation Evolution 300 and Evolution 600 User s Gui...

Page 30: ...nstrument Here is an example of a live status display Options window To access the Options window from the Main Menu press the Options function key 26 Evolution 300 and Evolution 600 User s Guide Thermo Electron Corporation ...

Page 31: ...e automatically printed at the end of each run This option uses the currently selected printer Toggled Off no automatic printout User must Print to obtain a printout if desired AutoSave Results Toggled On test results are automatically saved after each experiment in a user named file stored in either the internal instrument memory or on a disk When On the user must enter a name deed and start numb...

Page 32: ...e screen includes only the current wavelength Print Screen bmp Toggled On allows the user to create a bitmap of the current screen contents to be saved to a disk when the Print button is pressed Toggled Off printing function is directed to the installed printer Run Timers Enable up to four countdown timers to be incorporated into the method sequence User Log on Toggled On a password is required to...

Page 33: ...fault baseline is really needed before selecting this option Initialize Optics Re initializes optics without switching the instrument off then on Lifetime Initialization Performs a lifetime initialization Ensure that a lifetime initialization is really needed before selecting this option Wavelength Calibration Performs a wavelength calibration using the optional mercury lamp accessory Ensure that ...

Page 34: ...ssory Smart Accessories To access the accessory controls press the Accessories function key on the Main Menu page A pop up menu shows the accessories currently installed Here is an example Highlight the required accessory and press Enter to open the setup page When accessories are installed additional items related to the control of the accessories are added to the method parameter pages 30 Evolut...

Page 35: ...me on the spectrophotometer SmartStart The instrument can be configured so that frequently used methods are displayed on the SmartStart menu This then replaces the Main Menu as the Home page enabling common methods to be accessed conveniently from the start up screen Select files to be included in the SmartStart menu 1 Press Library 2 Highlight a file to be included in the SmartStart menu 3 Press ...

Page 36: ... SmartStart function key has been pressed Continue in this way until all required files have been chosen for the SmartStart menu 4 Press Home All of the selected items appear on the SmartStart Menu page 32 Evolution 300 and Evolution 600 User s Guide Thermo Electron Corporation ...

Page 37: ...the Library Press SmartStart Text entry Text items are entered using the text entry screen Here is an example of a text entry screen Navigate in the text entry screen Use the arrow keys to move left right and up down to highlight the required character To facilitate navigation the key action will wrap so that for example pressing the left arrow from A will move the highlight to M Thermo Electron C...

Page 38: ...ue of a numeric field highlight the item An Edit Value box appears Key in numbers with the numeric keypad taking note of any information such as limits displayed in the Edit Value box Press Enter when the numeric entry is complete or Cancel to terminate numeric entry to close the Edit Value box without making any changes When setting up a page requiring multiple numeric entries e g a Standards pag...

Page 39: ...ton on the instrument keypad Access the Library or disk from the Main Menu 1 Highlight the required field using the up down arrow keys and press Enter 2 Choose the required operation from the menu 3 Press Enter Load Method and Save Method function keys are available on application parameter pages Save Results function keys are available on all Results pages Note When a Load Function key is selecte...

Page 40: ... Filename Allows the user to enter a file name When filename has been entered press the Accept function key SmartStart Allows the user to place this method in the SmartStart menu when the SmartStart menu is set as the home page Method Name Allows the user to enter method name optional Drive Allows the user to select whether the method is saved to disk or instrument Library Press the Save function ...

Page 41: ...installed 2 Set up Method Parameter Function Scan Type Toggles between Standard and Intelliscan When Standard is selected the user is allowed to enter a scan speed When Intelliscan is selected the Speed item is used to select the Intelliscan mode which also determines the data interval Peak Table Sets up a Peak Table for peak operations that will be performed at run time Graph High Enter maximum v...

Page 42: ...ess the Save Method function key and follow the on screen instructions 5 Load a method from the instrument Library or from disk To do this press the Load Method function key or press Library 6 Remove any unwanted samples from sample compartment 7 Insert a suitable reference if required and close the lid of the sample compartment 8 Press Zero to measure a baseline 9 Open lid of the sample compartme...

Page 43: ... to open the Manipulate menu and choose the post run manipulation required For example press the View Results function key to see the results of Peak Table operations or post run manipulations Thermo Electron Corporation Evolution 300 and Evolution 600 User s Guide 39 ...

Page 44: ...ore items than are shown in the example depending on the way the software is configured and which accessories are installed 2 Highlight each parameter in turn and press Enter Parameter Function Data Mode Toggles between T and ABS Method Name Allows user to enter a method name Wavelength Selection Allows the user to select either Single Multiple or Serial Wavelength measurements Wavelength s Sets t...

Page 45: ...ngle use UVcalc Refer to Using UVcalc Wavelength Selection Use the table below to help you choose between the three different wavelength options Select this wavelength To do this Single Wavelength Measure each sample at one wavelength the same for each sample Multi Wavelength Measure each sample at up to 20 wavelengths the same for each sample Serial Wavelength Measure up to 9 samples at one wavel...

Page 46: ...sert the samples 7 Close the lid of the sample compartment and press Run 8 Press the View Results function key to see the Fixed Results Table From this page it is possible to save the results to the instrument Library or to disk 42 Evolution 300 and Evolution 600 User s Guide Thermo Electron Corporation ...

Page 47: ...ethod and press Enter Highlight each parameter in turn This will cause the item to toggle between two values or open a menu or open a numeric entry box Seven different data modes are available The number of wavelengths required will depend on the Data Mode Highlight Standards and press Enter to open the Standards Entry page Enter the concentration of each standard Press the Accept function key whe...

Page 48: ...ibrary or from disk 4 Clear the beams or insert suitable blank s and close the lid of the sample compartment 5 Press Zero to zero the instrument 6 Press the Calibrate function key Follow the on screen instructions which will depend on whether a cell programmer is in use and on the number of standards and replicates 7 View the graph and statistics This is done when the calibration is complete Chang...

Page 49: ...cs application Follow these steps to run a Kinetics application 1 Start the Kinetics application and press Enter The Kinetics Method page will open The menu may contain more items than are shown in the example depending on the way the software is configured and which accessories are installed Note The layout and composition of the Kinetics Method page will depend on the Measurement Mode selected T...

Page 50: ... each sample is measured in turn for the specified number of cycles The Cycle Time is the interval between measurements on the same cell Total Measurement Time is calculated from the number of cycles and the cycle time and is not editable in Parallel Kinetics mode 2 Set up the Method and press Enter Highlight each parameter in turn This will cause the item to toggle between two values or open a me...

Page 51: ...e Set up Graph High and Graph Low to accommodate the expected absorbance range The graph can be reconfigured post run if required Enter a factor and units if required Smoothing may be specified at run time or applied post run Save the new method into the instrument Library or to disk Press the Save Method function key and complete the save dialog as required 3 Load a previously saved method from t...

Page 52: ...cale smooth or change the absorbance display if required Select Original to return to the unmanipulated data 10 Set up the rate calculations Section allows up to 5 markers to be placed on the graph defining up to 4 contiguous time intervals over which the rate statistics will be calculated When the markers have been placed press the function key to exit from the Section Mode 11 Press the View Resu...

Page 53: ...plication 1 Start Multicomponent Analysis and press Enter The MCA method page will open The menu may contain more items than are shown in the example depending on the way the software is configured and which accessories are installed Thermo Electron Corporation Evolution 300 and Evolution 600 User s Guide 49 ...

Page 54: ...exactly the same method Each file must contain data for a single sample Use Clear Results between Fixed measurements Standards Transfers to the MCA standards page When Measure Standards Yes enter an ID and concentration for each standard When Measure Standards No load Standards as described above and enter a concentration for each standard Wavelengths When Measure Standards Yes enter the wavelengt...

Page 55: ...le to be measured at each of the wavelengths specified in the method The concentration of each component is then calculated from the calibration 7 Press the View Results function key to transfer to the MCA Results page The up down arrow keys can be used to view previous following results 8 Press the Save Results function key to save results to the instrument Library or to disk Biological tests Fol...

Page 56: ...ess Enter Choose the required test from the Nucleic Acid Tests sub menu The relevant test page opens The content and layout of the page will depend on the test selected the way the software is configured and which accessories are installed 3 Set up method parameters 52 Evolution 300 and Evolution 600 User s Guide Thermo Electron Corporation ...

Page 57: ...ht Display Protein and press Enter to toggle Yes No When Yes the protein factors are displayed 6 Edit the Dilution Multiplier if required 7 Edit the Units if required 8 When using a DNA method with scan press the Setup Scan function key This will access the Setup Scan page from which the start and stop wavelengths can be edited If the method has been edited it may be saved to the instrument Librar...

Page 58: ... 12 Press Run A message Measuring Sample appears Note Quartz cells MUST be used for all DNA measurements Glass or plastic cells are not suitable because they do not transmit at the wavelengths used The results page contains the results of the calculations and the Scan when applicable 13 Press the Save Results function key to save the results to the instrument Library or to disk 54 Evolution 300 an...

Page 59: ...t page opens The content and layout of the page will depend on the test selected the way the software is configured and which accessories are fitted The method parameters are documented values published in the literature Parameters may be edited if required The Standards Table contains recommended concentrations but can be edited as required 3 Edit the Units and enter High Low limits if required T...

Page 60: ...ructions to perform the calibration 9 Save the calibrated method to the instrument Library or to disk using a new file name Note A previously calibrated method can be loaded from the instrument Library or from disk Press the Load Method function key 10 Once the calibrated method is present place the sample in the cell holder and close the lid of the sample compartment 11 Press Run The results will...

Page 61: ...ment uses absorbance at 600nm to indicate the progress of cell growth in a sample The instrument does not perform any calculations or graph the data 1 Highlight Biological Tests on the Main Menu page The Biological Tests sub menu will open 2 Choose Cell Growth and press Enter Thermo Electron Corporation Evolution 300 and Evolution 600 User s Guide 57 ...

Page 62: ...the lid of the sample compartment 6 Press Run The sample number and absorbance are displayed on the Results page Use the Oligo calculator The oligonucleotide calculator determines the following data for a base sequence that you enter Number of bases Percent GC content Molecular weight Absorptivity extinction coefficient Conversion factor to be used in oligonucleotide measurements Tm for oligos up ...

Page 63: ...en 2 Choose Oligo Calculator and press Enter The Oligo Calculator page opens 3 Specify a base sequence You will need to enter a base sequence before you can run the oligonucleotide calculations Thermo Electron Corporation Evolution 300 and Evolution 600 User s Guide 59 ...

Page 64: ...ed base 4 Repeat these steps until you have specified the entire base sequence The displayed number of bases GC content DNA Molecular Weight DNA absorptivity extinction coefficient and factor will be updated as each new base is added to the sequence 60 Evolution 300 and Evolution 600 User s Guide Thermo Electron Corporation ...

Page 65: ...t calculator 6 Highlight parameter to be changed and press Enter A pop up menu will appear Here is an example Once all parameters have been set appropriately the relevant set of melting point predictions will be displayed Thermo Electron Corporation Evolution 300 and Evolution 600 User s Guide 61 ...

Page 66: ...es customized calculations to be incorporated into Scan Fixed and Quant methods 1 Highlight UVcalc on the selected application page and press Enter The equation selection page opens 62 Evolution 300 and Evolution 600 User s Guide Thermo Electron Corporation ...

Page 67: ... if you are starting a UVcalc calculation and press Enter The equation entry page for the selected equation opens with the Formula line highlighted 3 Press Enter to edit the formula The formula page opens Thermo Electron Corporation Evolution 300 and Evolution 600 User s Guide 63 ...

Page 68: ... Nested brackets are supported Operators Add subtract multiply divide spaces may be used if required 4 Use the arrow keys to move the highlight to the required element and press Enter Further options will be made available as required The back arrow and function keys may be used to edit the equation 5 When the equation is set up press the Accept function key The Cancel button enables you to exit w...

Page 69: ... the equation units for the results and to enable the result to be tested in order to establish whether it falls between defined limits Here is an example In each case highlight the item and pres Enter and then enter the test and or numbers in the usual way Thermo Electron Corporation Evolution 300 and Evolution 600 User s Guide 65 ...

Page 70: ...nding on the accessories installed Note Turn on the instrument at least an hour before starting performance qualification tests 1 From the Main Menu page press the Utilities function key 2 Highlight Instrument Verification and press Enter 66 Evolution 300 and Evolution 600 User s Guide Thermo Electron Corporation ...

Page 71: ...t function key to select it Here is an example 4 Press Run after selection s has been made Note The drift test will take at least an hour 5 Press the Summary function key to return to the Instrument Verification page Thermo Electron Corporation Evolution 300 and Evolution 600 User s Guide 67 ...

Page 72: ...an be loaded using the Load Results function key or by using the Library or disk menu items on the Main Menu page Further tests will become available when an instrument verification accessory CVC or Mercury lamp is installed Refer to the appropriate accessory manual 68 Evolution 300 and Evolution 600 User s Guide Thermo Electron Corporation ...

Page 73: ...e comprehensive on line Help system on the VISION CD or To start VISION you can either Double click on the VISION icon on the Windows desktop Select Start Programs VISIONpro VISIONpro or Start Programs VISIONsecurity VISIONsecurity as appropriate By default VISION uses the Com 1 port on the computer Thermo Electron Corporation Evolution 300 and Evolution 600 User s Guide 69 ...

Page 74: ......

Page 75: ... interior should be kept as dust free as possible The sample compartment cleaned regularly Wipe off any spilled chemicals immediately Cleaning instrument exterior The exterior and sample compartment of the instrument can be cleaned periodically as follows Caution Do not allow moisture to leak into the instrument 1 Switch off the spectrophotometer and disconnect from AC power source 2 Using a lint ...

Page 76: ...ernal printer paper 1 Open the printer housing To do this lift the cover and swing it away from the display Lift cover up swing back 2 Turn the paper release lever through 90 towards the display 3 Feed the paper shiny side down into the input slot Ensure that the paper emerges correctly through the printer and approximately 10cm 4 in has been pulled clear 72 Evolution 300 and Evolution 600 User s ...

Page 77: ...inter housing Light sources Evolution 600 only The tungsten halogen and deuterium lamps get very hot while the instrument is operating Before removing either lamp turn off the instrument and allow it to cool for 15 minutes CAUTION Thermo Electron Corporation Evolution 300 and Evolution 600 User s Guide 73 ...

Page 78: ...he wall outlet or power strip Warning The following lamp replacement procedures require removal of the lamp area cover Operation of the instrument with the optics cover removed exposes the operator and other observers to ultraviolet radiation which can damage the human eye and cause burning of the skin All personnel in the area must wear protective UV absorbing safety glasses 74 Evolution 300 and ...

Page 79: ...iver turn screws 2 turns counterclockwise b Rotate the lamp and pull upward until out of housing c Unplug inline connector 4 Insert new lamp Caution Never touch the bulb with your fingers Oil from your skin may cause the bulb to burn out quickly or explode a Plug in inline connector b Place lamp in lamp housing aligning hex screws c Using 2 5mm hex driver tighten screws 5 Replace rear cover 6 Plug...

Page 80: ...lding the lamp socket by the mounting tabs remove the copper spring clip c Unplug the tungsten halogen lamp from socket 4 Insert new bulb Caution Never touch the bulb with your fingers Oil from your skin will cause the bulb to burn out quickly or explode a Holding the lamp socket by the mounting tabs insert the bulb pins into the socket b Replace the copper spring clip c Replace the lamp socket en...

Page 81: ...pplicable must be reset from the controlling software Mercury lamp Evolution 300 and Evolution 600 Refer to the accessory user s guide for directions on replacing the mercury lamp Thermo Electron Corporation Evolution 300 and Evolution 600 User s Guide 77 ...

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