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Common Wavelengths for Fluorescence and Lumines-

SpectraMax GEMINI EM Operator’s Manual

B-3

Values in this table are based on the literature. You may wish to scan

your

uorochrome of interest in the SpectraMax GEMINI EM to

determine the optimal excitation and emission wavelengths for your
application.

Fluorescence

Luminescence

Emerald, Emerald II, Sapphire, Sapphire II, and Ruby are

trademarks of Tropix, Inc.

NOTE:

Fluorophore

Excitation

Wavelength

Emission

Wavelength

HPPA

320

405

4-MeU, NADH, NADPH

355

460

Biotinidinase

355

544

PKU

390

485

Green Fluorescent Protein

390

510

Attophos/Atto

uor

444

555

FITC

485

538

Ethidium Homodmer (DNA)

530

620

TRITC, Ethidium Bromide

544

590

Texas Red

584

612

TAMRA

547

580

Tryptophan

280

340

La Jolla Blue

695

705

Probe

Wavelength

Emerald and Emerald II

542

Sapphire and Sapphire II

461

Ruby

620

Содержание SpectraMax GEMINI EM

Страница 1: ...SpectraMax GEMINI EM Dual Scanning Microplate Spectrofluorometer Operator s Manual 1311 Orleans Drive Sunnyvale California 94089 Part 0112 0088 Rev A...

Страница 2: ...r company and product names are trademarks or registered trademarks of their respective owners Disclaimer Molecular Devices Corporation reserves the right to change its products and services at any ti...

Страница 3: ...oduct ex cept to the extent required by applicable law The Seller specifically excludes all express and implied warranties including without limitation any implied warranty that the products sold unde...

Страница 4: ...iv SpectraMax GEMINI EM Operator s Manual...

Страница 5: ...Temperature Regulation 1 12 Automix 1 13 Computer Control 1 13 Specifications 1 14 Glossary of Terms 1 16 Chapter 2 Installation 2 1 Installation Warnings 2 3 Installation Cautions 2 3 Unpacking 2 3 S...

Страница 6: ...pter 4 6 Removing the Drawer Adapter 4 7 Moving the SpectraMax GEMINI EM 4 7 Chapter 5 Troubleshooting 5 1 Error Codes and Resolutions 5 3 Opening the Drawer Manually 5 4 Appendix A Cables and Accesso...

Страница 7: ...re 1 7 Optimized Excitation and Emission Reading Wavelengths 1 10 Figure 2 1 View of Rear Panel 2 3 Figure 2 2 Adapter Inserted in Microplate Drawer 2 4 Figure 2 3 Microplate Drawer without Adapter 2...

Страница 8: ...viii SpectraMax GEMINI EM Operator s Manual Tables Table 3 1 Emission Cutoff Filter Default Settings 3 9 Table 5 1 Error Codes and Resolution 5 3...

Страница 9: ...ress down A note provides information that will help you properly execute an action or procedure Indicates an action or condition that could potentially dam age the instrument or one of its components...

Страница 10: ...x SpectraMax GEMINI EM Operator s Manual...

Страница 11: ...1 4 The Microplate Drawer 1 6 The Optical System 1 7 The Back Panel 1 8 Principles of Operation 1 9 Fluorescence 1 9 Luminescence 1 11 Functional Description 1 11 Modes of Operation 1 11 Temperature...

Страница 12: ...Chapter 1 1 2 SpectraMax GEMINI EM Operator s Manual...

Страница 13: ...d Microplates having 6 12 24 48 96 and 384 wells can be used in the SpectraMax GEMINI EM Top or bottom detection is available at the touch of a button Detection of species collected on membrane plates...

Страница 14: ...1 1 SpectraMax GEMINI EM The Control Panel Figure 1 2 Control Panel The control panel consists of an LCD and four pressure sensitive membrane keys which can be used to initiate and regulate the tempe...

Страница 15: ...t tem perature and actual temperature are shown on the front panel LCD display When the instrument is performing a kinetic or spectral scan the temperature keys on the front panel are disabled Opens o...

Страница 16: ...remain open for approxi mately ten seconds after which a beeping sound will alert you approximately two seconds before the drawer closes automatically The drawer will remain open longer during interva...

Страница 17: ...djacent wells 5 The light beam enters the well and if fluorescent molecules are present light of the emission wavelength is emitted back out to mirrors that focus it and send it to an optical bundle 6...

Страница 18: ...the fuse box containing two fuses that are required for operation Printer port Present but not used in this model of SpectraMax Serial port double shielded RS 232 for use with an external computer plu...

Страница 19: ...Figure 1 6 Excitation and Emission Spectra Because the intensity of the excitation light is usually many tens of thousands of times greater than that of the emitted light some type of spectral separa...

Страница 20: ...sess more accurately whether the emission is in fact the expected fluorophore or multiple fluorophores and not one generated by a variety of background sources or by contami nants Another benefit is t...

Страница 21: ...the microplate drawer The information contained in this section provides an overview of the instru ment capabilities For a complete description of the modes of operation how to choose instrument setti...

Страница 22: ...designed to regulate the temperature of the microplate chamber from 4 C above ambient to 45 C Upon power up when the incubator is off the temperature in the SpectraMax GEMINI EM microplate chamber is...

Страница 23: ...thin each well Automix must be selected before beginning a reading The actions associated with the Automix setting depend on the read mode chosen For endpoint mode enabling Automix will shake the plat...

Страница 24: ...gnal 3X STD DEV of baseline 8 0 fmol well FITC bottom read 3 0 fmol well FITC top read Luminescence Photometric Performance Wavelength range 250 850 nm Sensitivity signal 3X STD DEV of baseline 10 amo...

Страница 25: ...Storage temperature 20 to 65 C Operational altitude 2000 m Installation category II Pollution degree 2 System Validation Internal standards for fluorescence and wavelength Software Windows 95 98 NT 20...

Страница 26: ...tation light prior to the excitation monochromator In endpoint reads and emission spec tral scans selection of excitation filter is automatic In excitation spectral scans the user has the choice of no...

Страница 27: ...thermal changes the motion of subatomic particles or the excitation of an atomic system by radiation Parallel Port Windows A connection on a computer usually LPT1 where you plug in the cable for a pa...

Страница 28: ...Chapter 1 1 18 SpectraMax GEMINI EM Operator s Manual...

Страница 29: ...Chapter 2 Installation Installation Warnings 2 3 Installation Cautions 2 3 Unpacking 2 3 Setting Up the Instrument 2 3 Installing the Drawer Adapter 2 4 Removing the Drawer Adapter 2 5...

Страница 30: ...Chapter 2 2 2 SpectraMax GEMINI EM Operator s Manual...

Страница 31: ...in case there has also been damage to the instrument The SpectraMax GEMINI EM weighs approximately 35 pounds 16 kg and should be lifted with care It is recommended that two persons lift the instrumen...

Страница 32: ...he nearest objects or surfaces to ensure proper ventilation and cooling Installing the Drawer Adapter If you are reading standard 96 well or 384 well microplates from the top you will need to install...

Страница 33: ...or 48 well plates you will need to remove the adapter Incorrect insertion or removal of the adapter may cause dam age to the microplate drawer of the SpectraMax GEMINI EM 1 Turn power to the instrumen...

Страница 34: ...Chapter 2 2 6 SpectraMax GEMINI EM Operator s Manual...

Страница 35: ...urn the Instrument and Computer On3 3 Set the Temperature 3 3 Read the Microplate 3 3 Operation Overview 3 4 Optimizing Assays 3 4 Introduction 3 4 Using Spectral Scanning to Optimize Excitation and E...

Страница 36: ...Chapter 3 3 2 SpectraMax GEMINI EM Operator s Manual...

Страница 37: ...incubator Temp on off key The LCD dis play will indicate that temperature control is on and will show the set point and current temperature of the microplate chamber To change the temperature set poin...

Страница 38: ...this time 6 Load the prepared microplate into the drawer being sure to match well A1 with the A1 mark on upper left hand corner of the drawer 7 Using SoftMax Pro start the reading Optimizing Assays I...

Страница 39: ...PMT B Set the emission wavelength based on the tentative value from the lit erature or from a customary filter set used to measure your fluoro phore If the emission wavelength is not known select a te...

Страница 40: ...nd medium PMT B Set the excitation wavelength to the value determined in 2F above C Set the emission scan to start stop approximately 50 nm below or above the tentative emission value obtained from th...

Страница 41: ...Comments A In endpoint or kinetic fluorescence modes the Autofilter feature will generally select the same cutoff filter wavelength as will the above optimization method If desired however you may spe...

Страница 42: ...Chapter 3 3 8 SpectraMax GEMINI EM Operator s Manual Figure 3 2 Effects of Cutoff Filters on Fluorescein Emission was scanned from 490 to 560 nm excitation was fixed at 485 nm...

Страница 43: ...toff Selection Endpoint and Kinetic Modes l nm Emission l nm None 322 325 322 414 420 415 434 435 435 454 455 455 474 475 475 494 495 495 514 515 515 529 530 530 549 550 550 569 570 570 589 590 590 60...

Страница 44: ...Chapter 3 3 10 SpectraMax GEMINI EM Operator s Manual...

Страница 45: ...er 4 Maintenance Technical Support 4 3 Cleaning 4 3 Cleaning the Fan Filter 4 4 Changing the Fuses 4 4 Installing the Drawer Adapter 4 6 Removing the Drawer Adapter 4 7 Moving the SpectraMax GEMINI EM...

Страница 46: ...Chapter 4 4 2 SpectraMax GEMINI EM Operator s Manual...

Страница 47: ...cover or disassembly of any interior instrument component Removal of protective covers that are marked with the High Voltage warning symbol shown below can result in a safety hazard Cleaning Wear glov...

Страница 48: ...EMINI EM If power failed while the SpectraMax GEMINI EM was already on check that the power cord is not loose or disconnected and that power to the power outlet is functioning properly If these checks...

Страница 49: ...instru ment Figure 4 2 Removing the Fuse Box 6 When removed the fuse assembly will appear as shown in Figure 4 3 The holder inside contains two fuses 7 It is possible that only one of the fuses may h...

Страница 50: ...ll microplates from the top you will need to install the drawer adapter Incorrect insertion or removal of the adapter may cause damage to the microplate drawer of the SpectraMax GEMINI EM The corner c...

Страница 51: ...e steps The SpectraMax GEMINI EM weighs approximately 35 pounds 16 kilograms To avoid injury it is recommended that two people lift the instrument together using proper lifting techniques 1 Remove any...

Страница 52: ...Chapter 4 4 8 SpectraMax GEMINI EM Operator s Manual...

Страница 53: ...Chapter 5 Troubleshooting Error Codes and Resolutions 5 3 Opening the Drawer Manually 5 4...

Страница 54: ...Chapter 5 5 2 SpectraMax GEMINI EM Operator s Manual...

Страница 55: ...codes appear on the LCD display cycle the power to the instrument turn power off and then on again after a few seconds If the error message still appears and no specific note appears next to that erro...

Страница 56: ...instrument off and then on again If the drawer remains closed turn the incu bator off if it was on by pressing the Temp on off key If the drawer still remains closed try using a blunt flat object such...

Страница 57: ...Appendix A Cables and Accessories Cables A 3 Items Available for Use with the SpectraMax GEMINI EM A 3...

Страница 58: ...Appendix A A 2 SpectraMax GEMINI EM Operator s Manual...

Страница 59: ...uirements Serial Interface Cable contact Molecular Devices for specific pin out requirements Macintosh Male DB8 to male DB8 IBM Compatible Male DB8 to Female DB9 Items Available for Use with the Spect...

Страница 60: ...Appendix A A 4 SpectraMax GEMINI EM Operator s Manual...

Страница 61: ...Appendix B Common Wavelengths for Fluorescence and Luminescence Fluorescence B 3 Luminescence B 3...

Страница 62: ...Appendix B B 2 SpectraMax GEMINI EM Operator s Manual...

Страница 63: ...uminescence aEmerald Emerald II Sapphire Sapphire II and Ruby are trademarks of Tropix Inc NOTE Fluorophore Excitation Wavelength Emission Wavelength HPPA 320 405 4 MeU NADH NADPH 355 460 Biotinidinas...

Страница 64: ...Appendix B B 4 SpectraMax GEMINI EM Operator s Manual...

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