Typhoon Trio User`s Guide - GE Healthcare Life Sciences
Contents
1. d Fe r Link With Use Image Emission Filter PMT Laser Sensitivity zj i 520 BP 40 CY2 ECL Blue FAM 600 Blue 488 v Normal y Green 532 lt Red 633 T 2 580 BP 30 Cy3 TAMRA AlexaFluor546 v 600 Blu Normal v ECL Excitation r 3 szogP30cys Normal v ra a4 610 BP 30 Deep Purple SYPRO Ruby E y Normal v Channel Data Warning DAL At If non optimal filter laser or beamsplitter combinations are Auto Link selected a warning is displayed before scanning is initiated Disable Warning Auto Link Mode Sensitivity Carcel Figure 3 2 The Fluorescence Setup window for the Typhoon Trio instrument Important p3 4 Typhoon Trio User s 3 3 2 Selecting the emission filter You select an emission filter from the Emission Filter list for each scan you want to perform The Emission Filter list displays the filters table 3 1 that are installed in the Typhoon Trio instrument If you do not want to use an emission filter select None from the list The laser and emission filter combinations listed in the Fluorescence Setup window are based on the excitation and emission maxima of the selected fluorochrome For most gel microplate and array samples these combinations provide the best scanning results However for membrane samples selecting a laser and emission filter combination that is close to the excitation and emission maxima might n2. o o o o oooo oo 3 4 3 3 3 Selecting the laser ent ir ne Le Based 3 5 Chapter 4 Troubleshooting Typhoon Trio User s Guide Addendum iii Warning A Caution Important Warning A Preface About this addendum The Typhoon Trio User s Guide Addendum v5 0 provides an overview of the Typhoon Trio and the Typhoon Trio model instruments This addendum describes only the features and functions that differ from the Typhoon instrument models described in the Typhoon User s Guide v3 0 You should use this addendum in conjunction with the user s guide to learn how to operate and maintain the Typhoon Trio or the Typhoon Trio system Related publications In addition to the Typhoon Trio User s Guide Addendum Typhoon User s Guide and Typhoon Installation Instructions you should be familiar with the ImageQuant TL image analysis software Safety Chapter 2 in this addendum and chapter 2 in the Typhoon User s Guide provide important safety information that should be used when operating the Typhoon system Before using the Typhoon system read and understand the safety information thoroughly Special safety text Make sure you follow the precautionary statements presented in this guide Indicates a possibility of severe or fatal injury to the user or other persons if the precautions or instructions are not observed Indicates that damage to the instrument loss of data or invalid data could occur if t
3. e Multilabel excited mode Creates an image from a sample labeled with up to four fluorescent dyes The dyes can be excited by one or more of the four laser choices 3 2 Emission filters The Typhoon instrument contains a standard set of emission filters and can accommodate a total of 13 emission filters The instrument uses emission filters to reject reflected and scattered excitation light and background fluorescence while allowing the emitted light from the sample to pass through to the PMT Table 3 1 lists the standard set of emission filters used with fluorescence scanning Table 3 1 Standard set of emission filters for the Typhoon Trio and Typhoon Trio instruments Emission filter Description 520 nm band pass filter Transmits light between 500 nm and 540 nm and has a transmission peak 520 BP 40 centered at 520 nm Use this filter with fluorochromes such as Cy 2 ECL Plus and fluorescein 555 nm band pass filter Transmits light between 545 nm and 565 nm and has a transmission peak 555 BP 20 centered at 555 nm Use this filter with fluorochromes such as R6G and HEXTM 580 nm band pass filter Transmits light between 565 nm and 595 nm and has a transmission peak 580 BP 30 centered at 580 nm Use this filter with fluorochromes such as TAMRA and Cy3 610 nm band pass filter Transmits light between 595 nm and 625 nm and has a transmission peak 610 BP 30 centered
4. manual Now part of GE Healthcare imoenaton at wor G6 Typhoon Trio User s Guide Addendum Version 5 0 for Microsoft Windows Amersham ie eee Y Biosciences Cy ECF ECL Plus ImageQuant and Typhoon are trademarks of Amersham Biosciences Limited Amersham and Amersham Biosciences are trademarks of Amersham plc Deep Purple is a trademark of FluroTechnics Pty Ltd Deep Purple total protein stain is exclusively licensed to Amersham Biosciences from FluoroTechnics Pty Ltd Deep Purple total protein stain may only be used in life science research Ethernet is a trademark of Xerox Corporation HEX ROX and TAMRA are trademarks of Applera Corporation Microsoft and Windows are trademarks of Microsoft Corporation SYPRO is a trademark of Molecular Probes Inc The Typhoon instrument is covered by one or more of the following US patents 5 528 050 5 459 325 and foreign equivalents The Typhoon system is for research purposes only It is not intended or approved for diagnosis of disease in humans or animals All goods and services are sold subject to the terms and conditions of sale of the company within the Amersham group that supplies them A copy of these terms and conditions is available on request Amersham Biosciences UK Limited Amersham Place Little Chalfont Buckinghamshire HP7 9NA UK Amersham Biosciences AB SE 751 84 Uppsala Sweden Amersham Biosciences Corp 800 Centennial Avenue PO Box 1327 Piscataw
5. Typhoon Trio instrument The multislide tray allows you to scan up to 30 microarray slides at a time See the Typhoon Multislide User s Guide for a description of how to use the multislide tray 1 3 Scan acquisition modes You can scan samples on the Typhoon Trio or Typhoon Trio instrument using the following scan acquisition modes e Storage phosphor mode Creates images from samples labeled with radioisotopes using storage phosphor technology e Fluorescence mode Creates images from samples labeled with fluorescent dyes For details on the available excitation modes see chapter 3 e Chemiluminescence mode Creates images from chemiluminescent samples 1 4 Before using the system Before using the Typhoon Trio system you should become familiar with e Safety information It is imperative that you follow the precautions in this addendum and in the Typhoon User s Guide chapter 2 e Basic operational instructions See the Typhoon User s Guide e Maintenance information for the instrument and accessories See the Typhoon User s Guide Typhoon Trio User s Guide Addendum pl 3 Warnings A Chapter 2 Safety The operator of the Typhoon system is assumed to be trained in the correct operation of the system and the safety issues Throughout the Typhoon User s Guide and this addendum the word you refers to this trained operator Using controls making adjustments or performing procedures other than t
6. the safety interlocks on the sample lid or try to gain N access to the interior of the instrument through any other opening Exposure to laser light can cause injury For example viewing the laser light directly can cause blindness The labels shown in figure 2 1 are aff xed to the instrument and warn of exposure to laser light CAUTION DANGER Laser radiation when open and interlock defeated AVOID DIRECT EXPOSURE TO BEAM VORSICHT GEFAHR Laserstrahlung wenn offen und Verriegelung ausgeschaltet CAUTION DIREKTE BESTRAHLUNG VERMEIDEN CLASS 1 Laser Radiation ATTENTION LASER PRODUCT BE N este ces AV ID EXP URE au rayonnement laser si le dispositif de verrouillage a t Per IEC 825 TO BEAM tal International compliance Eye exposure Direct exposure Figure 2 1 Laser light warning labels p2 2 Typhoon Trio User s Guide Addendum Chapter 3 Fluorescence scanning This chapter describes the emission filters and the fluorescence setup for the Typhoon Trio and Typhoon Trio instruments The topics are e Excitation modes section 3 1 e Emission filters section 3 2 About the Fluorescence Setup window section 3 3 See the Typhoon User s Guide for details on the following About fluorescence scanning including how fluorescence works beamsplitters linking fluorescent scans multilabel experiments and common fluorochromes Setting up for fluor
7. at 610 nm Use this filter with fluorochromes such as Deep Purple total protein stain ROX EtBr SYPRO Red and SYPRO Ruby 670 nm band pass filter Transmits light between 655 nm and 685 nm and has a transmission peak 670 BP 30 centered at 670 nm Use this filter with fluorochromes such as Cy5 526 nm short pass filter Transmits light below 526 nm Using this filter with some applications such as 526 SP ECFTM plotting can reduce the background level 390 nm band pass filter This filter is used for storage phosphor scanning and is not normally used for 390 BP 100 fluorescence scanning See the Typhoon User s Guide for instructions on how to install and remove an emission optical filter p3 2 Typhoon Trio User s Guide Addendum 3 3 About the Fluorescence Setup window y AAA j 3 3 About the Fluorescence Setup window See the Typhoon User s Guide for details on how to start the Scanner Control software and set the scan parameters in the Scanner Control window You use the Fluorescence Setup window in Scanner Control to select the fluorescence parameters to use with the sample You can select from one through four sets of parameters to perform one through four scans This section describes the Emission Filter list and the Laser list in the Fluorescence Se
8. ay NJ 08855 USA Amersham Biosciences Europe GmbH Munzinger Strasse 9 D 79111 Freiburg Germany Amersham Biosciences SV Corp 928 East Arques Avenue Sunnyvale CA 94085 4520 USA Amersham plc 2004 All rights reserved September 2004 Table of contents Preface Aboutthis addendums iaa ne er Re V Related publicatiOris x 2 teria End eget etr bk e gu s V Safety 1 see A YE IO EVA Sn xa done raus V Sp cial safety Aext coe Hate VEO ears V Trainied ODeFator s n sack oce bs an wards ikea V ASSU tmiptloris s cuero Sa hls educ A ey DA e Ros que veg vi Sateby Staridards i 54r vise d UE Vtde VER Sei vi ASSISTANCE Ju d ux eh e dp Ee ase Lbs e E Ru eggs vi Chapter 1 Introduction dl Sit requirements us uoo ar Nr E RY DER 1 2 1 2 System components and optional accessories o 1 2 1 21 System COMPONeN S o oooooooooo lle 1 2 1 2 2 Optional accessories l l 1 3 1 3 CScan acquisition modeS s 4 mus ds SA EI te 1 3 14 Before using the system 2 2 eee 1 3 Chapter 2 Safety 21 Safety overview ur ar an ete D EEASG EE LE E 2 1 2 2 Lase rlight safety rc een ln 2 1 Chapter 3 Fluorescence scanning 3 1 EXCITATION modes oe retro o A Sg RR ken RR E 3 1 3 2 EMISSION MES on rere peces Ahad E ee e 3 2 3 3 About the Fluorescence Setup window 2 2 222 3 3 3 3 1 Displaying the Fluorescence Setup window 3 3 3 3 2 Selecting the emission filter
9. escence scanning including preparing and placing the sample in the instrument e Scanning a fluorescent sample including checking the instrument state selecting the scan parameters selecting the fluorescence parameters removing the sample from the instrument cleaning the glass platen and sample lid and analyzing or preprocessing the scanned image 3 1 Excitation modes The available excitation modes for fluorescence scanning are e Red excited mode Creates an image from a sample labeled or stained with a fluorescent dye or from a chemifluorescence sample that is excited at 633 nm e Green excited mode Creates an image from a sample labeled or stained with a fluorescent dye or a from chemifluorescence sample that is excited at 532 nm e Blue excited modes Blue 488 Creates an image from a sample labeled or stained with a fluorescent dye or from a chemifluorescence sample that is excited at 488 nm using full laser power This mode is recommended for samples that have low background levels Typhoon Trio User s Guide Addendum p3 1 Chapter 3 Fluorescence scanning ECL Excitation Creates an image from a sample labeled or stained with a fluorescent dye or from a chemifluorescence sample that is excited at 488 nm using reduced laser power This mode provides the optimum excitation for ECL Plus chemifluorescence blot samples and for samples with high background levels that can be excited at a lower laser power
10. he user fails to comply with the advice given Highlights information that is critical for optimal performance of the system Note Identifies items of general interest Trained operator The operator of the Typhoon system is assumed to be trained in the correct operation of the system and the safety issues Throughout the Typhoon Trio User s Guide Addendum and the Typhoon User s Guide the word you refers to this trained operator Typhoon Trio User s Guide Addendum v Preface Assumptions The software related instructions in this addendum assume you have basic computer skills You should be familiar with the Microsoft Windows graphical user interface If you do not have these skills refer to the documentation or the Help for Windows Safety standards The Typhoon instrument complies with CE and other applicable standards For the latest conformity information contact Technical Support See the Assistance section for contact information Assistance When calling for assistance be prepared to supply the serial number of your instrument For contact by phone or fax please use one of the numbers below Alternatively an updated list of contacts is available on the Web at www amershambiosciences com Amersham Biosciences is now part of GE Healthcare The GE Healthcare Web site is at www gehealthcare com vi Typhoon Trio User s Guide Addendum Asia Pacific Tel 852 2811 8693 Fax 852 2811 5251 Australas
11. hose specified in the Typhoon User s Guide or this addendum can result in hazardous exposure to laser light high voltage or moving parts Exposure to these hazards can cause severe injury or death 2 1 Safety overview Make sure you understand and follow the safety precautions in the Typhoon User s Guide including e General safety precautions Typhoon serial numbers and safety labels e Electrical safety e Pinching and general warnings e Hazardous materials precautions See section 2 2 of this addendum for laser light safety 2 2 Laser light safety The Typhoon Trio instrument is a Class I laser instrument that houses three Class IIIB lasers blue green and red Under the specified operating procedures the instrument does not allow operator exposure to laser light However the lasers are accessible in the interior of the instrument The blue laser has a specified maximum output of 27 mW at 488 nm in a diverging beam The green laser has a specified maximum output of 25 mW at 532 nm in a diverging beam The red laser has a specified maximum output of 13 mW at 633 nm in a diverging beam The safety interlocks in the Typhoon instrument are designed to prevent you from being exposed to the laser beams If you open the sample lid while the scanner is in operation the laser shutter closes and blocks the light from the lasers Typhoon Trio User s Guide Addendum p2 1 Chapter 2 Safety Warning Do not attempt to defeat
12. ia Tel 61 2 9899 0999 Fax 61 2 9899 7511 Austria Tel 01 576 0616 10 Fax 01 576 0616 27 Belgium Tel 0800 73 888 Fax 03 272 1637 Canada Tel 1 800 463 5800 Fax 1 800 567 1008 Central East and Southeast Europe Tel 43 1 982 3826 Fax 43 1 985 8327 Denmark Tel 45 16 2400 Fax 45 16 2424 Finland amp Baltics Tel 358 0 9 512 39 40 Fax 358 0 9 512 17 10 France Tel 01 69 35 67 00 Fax 01 69 41 96 77 Germany Tel 0761 4903 291 Fax 0761 4903 405 Italy Tel 02 27322 1 Fax 02 27302 212 Japan Tel 81 3 5331 9336 Fax 81 3 5331 9370 Assistance Latin America Tel 55 11 3933 7300 Fax 55 11 3933 7315 Middle East and Africa Tel 30 1 96 00 687 Fax 30 1 96 00 693 Netherlands Tel 0165 580 410 Fax 0165 580 401 Norway Tel 2318 5800 Fax 2318 6800 Portugal Tel 21 417 70 35 Fax 21 417 31 84 Russia amp other C LS amp N LS Tel 7 095 232 0250 956 1137 Fax 7 095 230 6377 Southeast Asia Tel 60 3 8024 2080 Fax 60 3 8024 2090 Spain Tel 93 594 49 50 Fax 93 594 49 55 Sweden Tel 018 612 1900 Fax 018 612 1910 Switzerland Tel 01 802 81 50 Fax 01 802 81 51 UK Tel 0800 616928 Fax 0800 616927 USA Tel 1 800 526 3593 Fax 1 877 295 8102 Typhoon Trio User s Guide Addendum vii Chapter 1 Introduction The Typhoon instrument is a variable mode imager that produces digital images of radioactive fluorescent or chemilumine
13. ng 10 80 relative humidity 1 2 System components and optional accessories 1 2 1 System components The Typhoon Trio system includes the Typhoon Trio or Typhoon Trio instrument Scanner Control software Typhoon Direct Instrument Access software Ethernet cables power cords and tools The Typhoon instrument connects to the computer using an Ethernet connection The Typhoon Trio instrument contains the following three internal lasers Blue laser A solid state laser with a specified minimum output of 20 mW at 488 nm in a diverging beam Green laser A green visible laser with a specified minimum output of 20 mW at 532 nm in a diverging beam Red laser A red visible laser with a specified minimum of 10 mW at 633 nm in a diverging beam The Scanner Control software which runs on the Microsoft Windows operating system contains the graphical interface that allows you to control the instrument In addition Internet Explorer is required for the Typhoon Direct Instrument Access software You use the Typhoon Direct Instrument Access software to perform several maintenance procedures which are described in the Typhoon User s Guide pl 2 Typhoon Trio User s Guide Addendum 1 3 Scan acquisition modes 1 2 2 Optional accessories See the Typhoon User s Guide for a description of the optional accessories for the storage phosphor and fluorescence scanning modes In addition the Typhoon multislide tray is available for the
14. ot provide the best results because of the high background interference For the best results you should test the membrane samples with different laser and emission filter combinations to determine the optimal combination Guide Addendum 3 3 About the Fluorescence Setup window 3 3 3 Selecting the laser When you select a filter Scanner Control automatically selects a default laser to use with the emission filter you selected However if desired you can select a laser other than the default If you select a laser other than the default and then click OK a message appears and asks you to confirm the selection The Typhoon Trio laser choices are e Green 532 Red 633 Blue 488 e ECL Excitation The laser power varies from one wavelength to another You might need to adjust the PMT voltage accordingly For a description of the laser excitation modes see section 3 1 Typhoon Trio User s Guide Addendum p3 5 Chapter 4 Troubleshooting For basic troubleshooting guidelines see the troubleshooting section ofthe Typhoon User s Guide In addition to the guidelines presented in the Typhoon User s Guide the following error message appears if the Typhoon Trio instrument has a problem communicating with the internal blue laser Failed Communicating with Internal Laser Unit If this message appears turn off the power switch on the right side of the Typhoon instrument wait a few seconds and then turn the instrumen
15. scent samples The Typhoon system consists of an instrument figure 1 1 instrument control software and a variety of accessories The Typhoon instrument is available in several models This addendum provides an overview of the Typhoon Trio and Typhoon Trio models The Typhoon Trio instrument contains three internal lasers blue green and red The Typhoon Trio is a Typhoon Trio instrument with the additional 10 um pixel size scanning resolution The topics in this chapter are e Site requirements section 1 1 e System components and optional accessories section 1 2 e Scan acquisition modes section 1 3 e Before using the system section 1 4 Figure 1 1 The Typhoon Trio instrument For a detailed description of how to use the Typhoon Trio system see the Typhoon User s Guide Typhoon Trio User s Guide Addendum pl 1 Chapter 1 Introduction 1 1 Site requirements Table 1 1 provides a summary of the Typhoon Trio instrument site requirements For additional site guidelines see the Typhoon Installation Instructions for the Microsoft Windows XP Workstation Table 1 1 Typhoon Trio system instrument specifications Parameter Value Power requirements 100 120 V 6 A 50 60 Hz 220 240 V 4 A 50 60 Hz Weight Approximately 160 kg 350 Ib Dimensions Width 118 11 cm 46 5 in Height 48 26cm 19 in Depth 77 47 cm 30 5 in Temperature 15 30 C 59 86 F Humidity Noncondensi
16. t back on If the error happens repeatedly contact Technical Support See Assistance in the preface for contact information Typhoon Trio User s Guide Addendum p4 1
17. tup window for the Typhoon Trio and Typhoon Trio instruments For a description of how to set the PMT and sensitivity and how to link scans see the Typhoon User s Guide 3 3 1 Displaying the Fluorescence Setup window In the Acquisition Mode area of the Typhoon Scanner Control window figure 3 1 select Fluorescence and then click Setup The Fluorescence Setup window appears figure 3 2 xj Eile Templates Tray Help r Scanner Information r Setup Typhoon Model Typhoon Trio Address 127 0 0 1 Serial Number 11772948 User Name A NN Ze ee ee a model pm SES m Acquisition Mode Template default I Ly alee R CE rj bd 0 Phosphor Mode l p E Best Sensitivity 4 BestResolution N M r Options L Orientation ex I Press Sample 5 Pixel size Focal Plane l 200 microns Y Platen H B User Comment n x D C B Image Analysis None M Ar 123456 7 8S 101112 1314015 16 1 18 1920 2122 7 DIGE File Naming Format Scanning Information Total Scan Time 14 Min 48 Sec 1 580BP30 Green 532nm Normal 600V Lines Per Channel 1500 Image Size per Channel 2 861 MB Chan Filter Laser Sensitivity PMT Splitter 2 670BP30 Red 633nm Normal 600V Figure 3 1 Typhoon Trio Scanner Control window Typhoon Trio User s Guide Addendum p3 3 Chapter 3 Fluorescence scanning Emission Filter list Fluorescence Setup for Typhoon Laser list Trio
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