Molecular Devices SpectraMax iD3 User Manual
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Molecular Devices SpectraMax iD3 User Manual

Multi-mode microplate reader
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SpectraMax iD3
Multi-Mode Microplate Reader
User Guide
5054747 H
January 2022

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  • Page 1 SpectraMax iD3 Multi-Mode Microplate Reader User Guide 5054747 H January 2022...
  • Page 2 For research use only. Not for use in diagnostic procedures. The trademarks mentioned herein are the property of Molecular Devices, LLC or their respective owners. These trademarks may not be used in any type of promotion or advertising without the prior written permission of Molecular Devices, LLC.

  • Page 3: Table Of Contents

    Contents Safety Information Warnings, Cautions, Notes, and Tips Chapter 1: Introduction Computer Integration Applications Environmental Control Optics Plate Controls Injectors Chapter 2: Setting Up the Instrument Package Contents Unpacking the Instrument Removing the Transport Locks Connecting Instrument Cables Assembling and Maintaining Injectors Chapter 3: Getting Started Welcome Defining Date, Time, and Global Read Settings...
  • Page 4 SpectraMax iD3 Multi-Mode Microplate Reader User Guide Read Result Library Chapter 6: Read Modes and Read Types Absorbance Read Mode Fluorescence Intensity Read Mode Luminescence Read Mode Chapter 7: Maintenance and Troubleshooting Preventive Maintenance Cleaning the Instrument Cleaning Injectors and Accessories...

  • Page 5: Safety Information

    Safety Information Safety Information Information about the safe use of the instrument from Molecular Devices® includes an understanding of the user-attention statements in this guide, the safety labels on the instrument, precautions to follow before you operate the instrument, and precautions to follow while you operate the instrument.
  • Page 6 SpectraMax iD3 Multi-Mode Microplate Reader User Guide Symbols on the Instrument Each safety label on the instrument contains an alert symbol that indicates the type of potential safety hazard. Symbol Indication Consult the product documentation. Potential lifting hazard. To prevent injury, use a minimum of two people to lift the instrument.
  • Page 7 For products under the requirement of the WEEE directive, contact your dealer or local Molecular Devices office for the procedures to facilitate the proper collection, treatment, recovery, recycling, and safe disposal of the device.
  • Page 8 Electrical Safety To prevent electrical injuries and property damage, inspect all electrical equipment before use and report all electrical deficiencies. Contact Molecular Devices technical support for equipment service that requires the removal of covers or panels. WARNING! HIGH VOLTAGE.
  • Page 9 The instrument contains moving parts that can cause injury. Under normal conditions, the instrument is designed to protect you from these moving parts. WARNING! If the instrument is used in a manner not specified by Molecular Devices, the protection provided by the equipment might be impaired. To prevent injury: Never try to exchange labware, reagents, or tools while the instrument is operating.
  • Page 10 SpectraMax iD3 Multi-Mode Microplate Reader User Guide 5054747 H...

  • Page 11: Chapter 1: Introduction

    Chapter 1: Introduction The SpectraMax® iD3 Multi-Mode Microplate Reader from Molecular Devices is a monochromator-based, multi-mode plate reader. The touchscreen interface provides integrated instrument control, data display, and the ability to export results over your network for statistical data analysis.
  • Page 12 SpectraMax iD3 Multi-Mode Microplate Reader User Guide In the Ribbon, on the GxP tab, users with appropriate permission can use the following icons to lock and unlock the instrument touchscreen: Click GxP Mode On to lock the instrument touchscreen and operate the instrument from the computer running the SoftMax Pro Software in GxP mode.
  • Page 13 The update file has the following naming convention: iDx_update_n_n_nnnnnn.mdup iDx_update: Use the SoftMax Touch Software update files to update the software in both the SpectraMax iD3 AND the SpectraMax iD5 instruments. The update is intelligent and knows which instrument software to update.
  • Page 14 Available Services to display the list of SpectraMax iD3 and SpectraMax iD5 instruments on your intranet and/or the instrument to which you connect the computer through an Ethernet cable.

  • Page 15: Applications

    Application notes with specific application protocol suggestions are in the Information Center and Knowledge Base on the Molecular Devices web site at www.moleculardevices.com. Environmental Control The instrument can maintain the temperature inside the plate chamber at 5°C (9°F) above ambient to 66°C (150°F).

  • Page 16: Plate Controls

    CAUTION! SpectraCuvette Adapters without a sticker have a plate height of 24 mm and cannot be used in the SpectraMax iD3 or SpectraMax iD5. To prevent damage to the instrument, the height of the plate must not exceed 22 mm, including the lid if the plate is lidded.

  • Page 17: Chapter 2: Setting Up The Instrument

    Chapter 2: Setting Up the Instrument Before you unpack and setup the SpectraMax iD3, prepare a flat work area that has sufficient space for the instrument and required cables. See Instrument Specifications on page 101. All software required to run basic non-injector reads is installed in the instrument and is accessible from the touchscreen.
  • Page 18 SpectraMax iD3 Multi-Mode Microplate Reader User Guide When your instrument has the SpectraMax Injector System the package also contains the following. Package Contents for Injectors Illustration Part Number Description 5055247 Injector Nozzle 5044164 Tubing 5055251 Bottle Holder 5044165 Bottle Adapters...

  • Page 19: Unpacking The Instrument

    Chapter 2: Setting Up the Instrument Unpacking the Instrument The packaging is designed to protect the instrument during transportation. Transport locks are placed on the transport slide and the plate drawer to protect the instrument from damage during shipment. You must remove the transport locks before you power on the instrument.
  • Page 20 SpectraMax iD3 Multi-Mode Microplate Reader User Guide 3. Grasp the handle on the cardboard and slide the instrument out of the box. 4. Remove the accessories tool box. CAUTION! Keep the instrument upright and level when lifting. Do not tip or shake the instrument to prevent damage to the instrument.

  • Page 21: Removing The Transport Locks

    Chapter 2: Setting Up the Instrument Removing the Transport Locks CAUTION! The instrument can be damaged if the transport locks are not removed before the instrument is powered on. Transport locks are placed on the transport slide and the plate drawer to protect the instrument during shipment.
  • Page 22 SpectraMax iD3 Multi-Mode Microplate Reader User Guide 2. Use the provided 3.0 mm Holex HEXAGON ballhead bolt driver to loosen the screw located inside the Transport Lock opening until you feel the spring release. This unlocks the transport slide. The screw remains inside the instrument. The screw is spring mounted and cannot get lost within the instrument.
  • Page 23 Chapter 2: Setting Up the Instrument 5. Use the provided 2.0 mm hex key to loosen screw #1 in the upper-left corner of the transport lock until the lock disconnects from the instrument frame. The screw has a retaining washer that prevents it from being removed from the lock. Tip: After you loosen screw #1, pull the plate drawer slightly out of the instrument to hold the chamber door open.

  • Page 24: Connecting Instrument Cables

    SpectraMax iD3 Multi-Mode Microplate Reader User Guide Connecting Instrument Cables The power cord and Ethernet cable connect to the ports on the rear of the instrument. The Ethernet port allows you to connect the instrument to your intranet (like a printer) or to connect the instrument directly to a computer.

  • Page 25: Assembling And Maintaining Injectors

    Chapter 2: Setting Up the Instrument Assembling and Maintaining Injectors The SpectraMax Injector System is an optional addition to the base instrument configuration. If there is no handle on the instrument right hood, your instrument does not have injectors and the right hood is sealed shut.
  • Page 26 SpectraMax iD3 Multi-Mode Microplate Reader User Guide Injector Tubing Each injector has an injector tube. The tubing line connects to an injector tip on one end and a snorkel on the other end. From the tip, the tube passes around the injector pump to the snorkel.
  • Page 27 Chapter 2: Setting Up the Instrument To install or replace the injector tubing: 1. Move the injector arm away from the instrument and insert the nozzle into the injector arm. Do the following for both injectors. 2. Insert the snorkel into the bottle and then insert the snorkel into the snorkel clamp. Clamp for injector 1 is on the left and clamp for injector 2 is on the right.
  • Page 28 SpectraMax iD3 Multi-Mode Microplate Reader User Guide 4. Slide the tip into the nozzle, the slots are labeled 1 and 2 for injector 1 and injector 2. 5. Use the black knob to move the nozzle back to the rear of the injector space. Align the nozzle with the opening and press straight down until you feel it snap into place.

  • Page 29: Chapter 3: Getting Started

    Chapter 3: Getting Started All software required to run basic non-injector reads is installed in the instrument and is accessible from the touchscreen. You must use a computer running the SoftMax Pro Software to operate the instrument for advanced acquisition settings and for protocols that use the SpectraMax Injector System.

  • Page 30: Welcome

    SpectraMax iD3 Multi-Mode Microplate Reader User Guide Welcome The Welcome page provides a list of the instrument features and allows you to select a user. Tap each of the features to view the help topic related to each feature. - Tap to hide the features.

  • Page 31: Defining Date, Time, And Global Read Settings

    Chapter 3: Getting Started Defining Date, Time, and Global Read Settings After you log in, you use the Maintenance page to define the system date/time and the global read settings that apply to all users of the instrument. To Defining Date, Time, and Global Read Settings: 1.

  • Page 32: Getting The Instrument On Your Network

    SpectraMax iD3 Multi-Mode Microplate Reader User Guide Getting the Instrument on Your Network After you connect the Ethernet cable from the instrument to a networked wall outlet and after you power on the instrument, the instrument should automatically have an IP address assigned.

  • Page 33: Maintaining Users

    Chapter 3: Getting Started Maintaining Users The Admin user manages the user accounts on the instrument. When you log in as the Admin, the Maintenance page contains a Users tab. The Admin user can create user accounts to require a PIN and/or an NFC tag. If a user forgets their PIN, the Admin assigns the user a new PIN that they can use to log in.
  • Page 34 SpectraMax iD3 Multi-Mode Microplate Reader User Guide 3. From the icons on the left, tap to display the Maintenance page and tap Users to display the Users tab, if needed. 4. Tap Create New User to display the New User dialog where you add a user.

  • Page 35: Chapter 4: Home Page

    Chapter 4: Home Page The Home page displays your favorite protocols, your recent protocols, and your result information. From the Welcome page, tap a user to display the Home page. Your login credentials determine what protocol information displays. See Welcome on page The following icon are added to left side of the page: - Return to your Home page.

  • Page 36: Changing Pin/Logout

    SpectraMax iD3 Multi-Mode Microplate Reader User Guide Changing PIN/Logout Use the Change PIN/Logout dialog to change your PIN and to logout. The Change PIN option does not appear for users that do not have a PIN. Note: If your account does not have a PIN and you want to add a PIN, use the Admin account to update your user account.

  • Page 37: Quick Start And Favorites

    Chapter 4: Home Page Reset Admin PIN If you forget the Admin user PIN, you can reset the Admin user PIN to the default 0000. The unlock code is the last four digits of the instrument serial number found on the Maintenance - System Information page and the sticker on the back of the instrument.

  • Page 38: Protocol Libraries

    SpectraMax iD3 Multi-Mode Microplate Reader User Guide Protocol Libraries Use the protocol libraries to access pre-defined protocol files that have settings but no data. The Home page provides access to the Standard library that contains the protocols included with the instrument software and the My Protocols library that contains the protocols you save.
  • Page 39 Chapter 4: Home Page My Protocols Library Tap My Protocols above the protocol list to display your My Protocols library that contains protocols associated to your user account. My Protocols protocols are only visible to you. You can add, export, copy, and delete your protocols. To manage the protocols in your My Protocol library: Tap a protocol to display the protocol settings.

  • Page 40: External Shake Settings

    SpectraMax iD3 Multi-Mode Microplate Reader User Guide External Shake Settings Use the external shake feature to shake a plate outside of the instrument. This shake process is independent of a protocol. When you create a protocol, the Settings page allows you to define how to shake the plate as part of the protocol.

  • Page 41: Temperature Settings

    Chapter 4: Home Page Temperature Settings The temperature inside the plate chamber can be maintained at 5°C (9°F) above ambient to 66°C (150.8°F). The temperature sensors detect the temperature of the air inside the chamber, not the temperature of the samples in the plate. If you use the instrument to warm, the samples, use a seal or lid on the plate to prevent evaporation of the sample.

  • Page 42: Maintaining Injectors

    SpectraMax iD3 Multi-Mode Microplate Reader User Guide Maintaining Injectors Use the Injector Maintenance page to wash, prime, and calibrate the injectors and to view the status of the amount of liquid that has been dispensed through the tubing to help you determine when to replace the tubing.
  • Page 43 Chapter 4: Home Page Tip: Instead of switching the bottles in and out of the bottle holder between the solutions, you can put both snorkels into one bottle and run each solution through the tubing for both injectors at the same time. To wash the injector tubing: 1.
  • Page 44 SpectraMax iD3 Multi-Mode Microplate Reader User Guide 12. Tap: to simultaneously run both injectors for the wash. Start Wash to dispense 900 µl for the wash. Manual to enter the amount of liquid to dispense then tap Start. 13. After the first wash step completes, move the snorkel or snorkels to the bottle on the right.
  • Page 45 Chapter 4: Home Page Priming Injector Tubing Before you run a read with the injectors, prime the injector tubing with the reagent that you use for the experiment. You can use the predefined prime process that dispenses 300 µl or you can use the Manual section to define how much solution to dispense, to do reverse prime, and to do air aspiration steps.
  • Page 46 SpectraMax iD3 Multi-Mode Microplate Reader User Guide 9. Fill the bottles with enough reagent for your experiment plus at least 2 mL to account for the prime operation and the quick-prime operation before the plate is read, and for the dead volume in the bottle and the tubing. Place the bottle for injector 1 on the left and the bottle for injector 2 on the right.
  • Page 47 Chapter 4: Home Page Injector Waste Plate and Strip Wells The waste plate captures excess liquid during the wash and prime operations. You use the strip wells during the quick-prime of the injectors that occurs when you start a read with injectors. Note: Make sure that the waste plate and strip wells are empty before you insert them.
  • Page 48 SpectraMax iD3 Multi-Mode Microplate Reader User Guide Injector Bottles The bottle holder holds two bottles that correspond with the two injectors. Fill the bottles with enough reagent for your experiment plus at least 2 mL to account for the prime operation and the quick-prime operation that occur before the plate is read, and for the dead volume in the bottle and the tubing.
  • Page 49 Chapter 4: Home Page 6. Slide the snorkel all the way down into the bottle and then slide the snorkel tube into the open end of the snorkel clamp. The bottle holder is slightly tilted toward one corner. To extract the maximum amount of liquid from the bottle, place the end of the snorkel in the lowest point that is located in the corner of the bottle closest to the closed end of the snorkel clamp.
  • Page 50 SpectraMax iD3 Multi-Mode Microplate Reader User Guide 9. Position the injector arm over the waste plate. Note: The injector arm does not line up with the hole in the waste plate. The instrument moves the waste plate to the proper position when you tap the Start buttons.
  • Page 51 Chapter 4: Home Page 39. Repeat the steps in the Calibration wizard for injector 2. 40. After you complete the Calibration wizard for injector 2, remove the plate from the carrier, remove the snorkels from the bottles, remove the bottles from the bottle holder, tap close the plate drawer, lift the tube stabilizer lids over the injector pumps, and then close the instrument right hood.

  • Page 52: Maintenance Page

    SpectraMax iD3 Multi-Mode Microplate Reader User Guide Maintenance Page All users can use the Maintenance page to view instrument information, manage reader settings, manage system settings, and to do support tasks. The Admin user can use the Maintenance page to manage users.

  • Page 53: Loading And Unloading Plates

    Chapter 4: Home Page Loading and Unloading Plates The icons on the left include to load or unload a plate: 1. Tap to move the plate drawer outside of the instrument. 2. Place the plate onto the plate carrier or remove the plate from the plate carrier. Note: Place the plate on the plate carrier in landscape orientation with well A1 in the left corner closest to the touchscreen.
  • Page 54 SpectraMax iD3 Multi-Mode Microplate Reader User Guide 5054747 H...

  • Page 55: Chapter 5: Protocols

    Tap Standard or My Protocols to select protocols stored in the file system. Application notes with specific application protocol suggestions can be found in the Information Center and the Knowledge Base on the Molecular Devices web site at www.moleculardevices.com. Viewing Protocol Settings Use the protocol settings page to manage the name of the protocol file, to select the plate for which to define settings, and to start a read.
  • Page 56 SpectraMax iD3 Multi-Mode Microplate Reader User Guide 2. To name the protocol: From the My Protocols library, tap to display the Create New Protocol dialog. From an existing protocol, tap <protocol name> to rename the protocol on the Rename Protocol dialog.

  • Page 57: Defining Protocol Settings

    Chapter 5: Protocols Defining Protocol Settings Use the plate-specific settings page to define the settings for each plate in the protocol. Settings vary depending upon the read mode and read type you select. Do the following for each plate in the protocol to define the plate format/plate type, read mode/read type, and other settings: 1.
  • Page 58 SpectraMax iD3 Multi-Mode Microplate Reader User Guide 4. The read mode and read type appear on the upper left side of the page. Tap <read mode/read type> to select the read mode and read type in the Read Mode/Type dialog.
  • Page 59 CAUTION! SpectraCuvette Adapters without a sticker have a plate height of 24 mm and cannot be used in the SpectraMax iD3 or SpectraMax iD5. To prevent damage to the instrument, the height of the plate must not exceed 22 mm, including the lid if the plate is lidded.
  • Page 60 SpectraMax iD3 Multi-Mode Microplate Reader User Guide Read Mode and Read Type Settings Use the Read Mode/Type dialog to set the protocol read mode and read type. After you select the read mode and read type on this dialog, the read mode/type displays at the top left on the plate-specific settings page.
  • Page 61 Chapter 5: Protocols 3. Tap a read type. - Endpoint - Kinetic - Well Scan - Spectrum Read Area Settings Use the plate-specific settings page to define the read area. All read mode read type combinations have a Read Area tab on the left to allow you to define which wells on the plate to read.
  • Page 62 SpectraMax iD3 Multi-Mode Microplate Reader User Guide To define which wells on the plate to read: 1. Start at the bottom of the tab. Select a Read Order: Select Row to read each row in sequence. Select Column to read each column in sequence.
  • Page 63 Chapter 5: Protocols Wavelength Settings Use the plate-specific settings page to define the wavelength. All read mode read type combinations have a Wavelength tab on the left that allows you to define which wavelengths to use for the plate read. The read mode and read type setting determines which wavelength settings are applicable.
  • Page 64 SpectraMax iD3 Multi-Mode Microplate Reader User Guide Absorbance - Spectrum Wavelength settings for the Absorbance mode with the Spectrum type: (Start and stop range from 230 - 1000 nm.) 1. Tap the Start and enter the excitation start wavelength. 2. Tap the Stop and enter the stop wavelength value.
  • Page 65 Chapter 5: Protocols Luminescence - Well Scan Wavelength settings for the Luminescence mode with the Well Scan type: Set the All Wavelength to use all wavelengths. 1. Set the All Wavelength to define wavelength settings. 2. Tap the wavelength and enter the wavelength value. Luminescence - Spectrum Wavelength settings for the Luminescence mode with the Spectrum type: 1.
  • Page 66 SpectraMax iD3 Multi-Mode Microplate Reader User Guide Detection Settings Use the plate-specific settings page to define the detection settings. All read mode read type combinations have a Detection tab on the left. The read mode setting determines which detection settings are applicable.
  • Page 67 Chapter 5: Protocols Fluorescence Mode Detection Settings Detection settings for the Fluorescence read mode: 1. Tap the PMT Gain drop-down: Select Auto to have the instrument adjust the PMT voltage automatically for varying concentrations of samples in the plate (not available for Kinetic type). Select High for samples that have lower concentration (dim samples).
  • Page 68 SpectraMax iD3 Multi-Mode Microplate Reader User Guide Shake Settings Use the plate-specific settings page to define the plate shake settings. All read mode read type combinations have a Shake tab on the left. To define how to shake the plate: Set Before First Read to not shake the plate as a part of the read.
  • Page 69 Chapter 5: Protocols Timing Settings For Kinetic read types, use the plate-specific settings page to define timing settings. All read mode Kinetic read types have a Timing tab on the left. The instrument calculates the number of reads based on the value you enter for the Total Run Time and the Interval.

  • Page 70: Read

    SpectraMax iD3 Multi-Mode Microplate Reader User Guide Read After you define the protocol settings for your read, tap Read to display a status bar. When you log in to the instrument as a user with PIN access, you can tap Lock Screen to prevent other users from interrupting your read.

  • Page 71: Result Library

    Chapter 5: Protocols Result Library The Result library displays the list of results for your reads. From the Results side of the Home page, tap View All to display the Result library. You can tap a result in your Most Recent results list to go directly to the Manage Results page. See Managing Results on page To use the Result library:...
  • Page 72 SpectraMax iD3 Multi-Mode Microplate Reader User Guide Spectrum Read Type Results Depending on the read mode, a Spectrum type raw absorbance, fluorescence, or luminescence data displays optical density (OD), % Transmittance (%T), relative fluorescence units (RFU), or relative luminescence units (RLU) across a spectrum of wavelengths that display as a plot.
  • Page 73 Chapter 5: Protocols Compare Wells Some results allow you to compare data in the wells. To compare data in wells: 1. Tap to compare the results in several wells. The icon spins and an additional icon appears. 2. Tap up to four wells to compare. Each well you select displays a shaded background. 3.
  • Page 74 SpectraMax iD3 Multi-Mode Microplate Reader User Guide Well Specific Linear Results Tap a single well in the result or select to compare multiple wells to display a view of a single well with the one result or the compared results.
  • Page 75 Chapter 5: Protocols Exporting Result Data When you connect the instrument to your intranet or you connect the instrument directly to a computer with an Ethernet cable, you must install the QuickSync Tool on the computer to which you want to export data. See Installing the QuickSync Tool on page The Read Settings tab on the Maintenance page provides an Export Excel option that allows the instrument to automatically export raw protocol data for further analysis.
  • Page 76 4. Right-click in the tray to display a menu and select Available Services to display the list of SpectraMax iD3 and SpectraMax iD5 instruments on your intranet and/or the instrument to which you connect the computer through an Ethernet cable.

  • Page 77: Chapter 6: Read Modes And Read Types

    Luminescence (LUM) read modes. This chapter describes these read modes and their associated read types. Application notes with specific application protocol suggestions can be found in the Information Center and the Knowledge Base on the Molecular Devices web site at www.moleculardevices.com. Read Types...
  • Page 78 SpectraMax iD3 Multi-Mode Microplate Reader User Guide Well Scan Read Type A Well Scan type can read at more than one location within a well. A Well Scan read takes one or more reads of a single well of a plate on an evenly spaced grid inside of each well at single or multiple wavelengths.

  • Page 79: Absorbance Read Mode

    Chapter 6: Read Modes and Read Types Absorbance Read Mode The instrument uses the Absorbance (ABS) read mode to measure the Optical Density (OD) of the sample solutions. Absorbance is the quantity of light absorbed by a solution. To measure absorbance accurately, it is necessary to eliminate light scatter.
  • Page 80 SpectraMax iD3 Multi-Mode Microplate Reader User Guide Applications of Absorbance Absorbance-based detection is commonly used to evaluate changes in color or turbidity, permitting widespread use including ELISAs, protein quantitation, endotoxin assays, and cytotoxicity assays. With absorbance readers that are capable of measuring in the ultraviolet (UV) range, the concentration of nucleic acids (DNA and RNA) can be found using their molar extinction coefficients.
  • Page 81 Chapter 6: Read Modes and Read Types PathCheck technology normalizes the data acquired from an absorbance read mode endpoint read type to a 1 cm pathlength, correcting the OD for each well to the value expected if the sample were read in a 1 cm cuvette. The instrument uses the factory installed water constant to obtain the 1 cm values.
  • Page 82 SpectraMax iD3 Multi-Mode Microplate Reader User Guide Note: It is important that you put water in the wells and do not read a dry plate for the Plate Background OD. A dry plate has a slightly higher OD value than a water filled plate because of differences in refractive indices.

  • Page 83: Fluorescence Intensity Read Mode

    Chapter 6: Read Modes and Read Types Fluorescence Intensity Read Mode Fluorescence occurs when absorbed light is re-radiated at a longer wavelength. In the Fluorescence Intensity (FL) read mode, the instrument measures the intensity of the re-radiated light and expresses the result in Relative Fluorescence Units (RFU). The governing equation for fluorescence is: Fluorescence = extinction coefficient ×...
  • Page 84 SpectraMax iD3 Multi-Mode Microplate Reader User Guide The Spectral Optimization wizard in the SoftMax Pro Software provides the best method to maximize the signal to background window (S-B)/B while minimizing the optimization time. Excitation maximum of fluorophore Emission maximum of fluorophore...
  • Page 85 Chapter 6: Read Modes and Read Types Applications of Fluorescence Intensity Fluorescence intensity is used widely in applications such as fluorescent ELISAs, protein assays, nucleic acid quantitation, reporter gene assays, cell viability, cell proliferation, and cytotoxicity. One more major application is to study the kinetics of ion release. Some assays use a fluorescent label to selectively attach to certain compounds.
  • Page 86 SpectraMax iD3 Multi-Mode Microplate Reader User Guide Linearity and the Linear Dynamic Range Within a wide range at moderately high concentrations, blanked raw data is proportional to the quantity of label in a sample. The linear dynamic range (LDR) is defined by:...

  • Page 87: Luminescence Read Mode

    Chapter 6: Read Modes and Read Types Luminescence Read Mode Luminescence is the emission of light by processes that derive energy from essentially non- thermal changes, the motion of subatomic particles, or the excitation of an atomic system by radiation. Luminescence detection relies on the production of light from a chemical reaction in a sample.
  • Page 88 SpectraMax iD3 Multi-Mode Microplate Reader User Guide Analyzing Luminescence Data The conversion rate of photons to counts is individual for each reader. Therefore, raw data from the same plate can seem significantly different from one instrument to the next. Also, the data format used by other manufacturers might not be counts per second and can be different by several orders of magnitude.
  • Page 89 Chapter 6: Read Modes and Read Types Z´ is the standard statistical parameter in the high-throughput screening community to measure the quality of a screening assay independent of test compounds. Use this as a measure of the signal separation between the positive controls and the negative controls in an assay. Use the following formula to determine the value of Z´: 3(SD ) + 3(SD...
  • Page 90 SpectraMax iD3 Multi-Mode Microplate Reader User Guide 5054747 H...

  • Page 91: Chapter 7: Maintenance And Troubleshooting

    Chapter 7: Maintenance and Troubleshooting Perform only the maintenance tasks described in this guide. Contact a Molecular Devices service engineer to inspect and perform a preventive maintenance service on the instrument each year. See Obtaining Support on page 100. Before operating the instrument or performing maintenance operations, make sure you are familiar with the safety information in this guide.

  • Page 92: Cleaning Injectors And Accessories

    SpectraMax iD3 Multi-Mode Microplate Reader User Guide Clean Touchscreen The touchscreen is made of coated glass. The coating that covers the glass is soft and can be scratched. Do not touch, push or rub the touchscreen with anything harder than an HB pencil lead.

  • Page 93: Troubleshooting

    Review the Chemical and Biological Safety information contained in this guide. See Chemical and Biological Safety on page Do only the maintenance described in this guide. Maintenance procedures other than those specified in this guide must be done by qualified Molecular Devices personnel only. See Obtaining Support on page 100.
  • Page 94 If these checks fail to remedy the loss of power, replace the fuses. You can obtain replacement fuses from Molecular Devices. For fuse specifications and part numbers, see Physical...

  • Page 95: Before You Move The Instrument

    Chapter 7: Maintenance and Troubleshooting Before You Move the Instrument When you move the instrument from one location to a new location, there are several things you must do before you power off the instrument. This procedure requires the following tool included in the accessories tool box: WARNING! LIFTING HAZARD.
  • Page 96 SpectraMax iD3 Multi-Mode Microplate Reader User Guide 6. On the rear of the instrument, remove the black cover from the travel lock opening in the center. 7. Insert the 3.0 mm Holex HEXAGON ballhead bolt driver into the Transport Lock opening and tighten the interior screw into the transport slide.
  • Page 97 Chapter 7: Maintenance and Troubleshooting Installing Transport Locks Before you move or pack the instrument, do all the steps in Before You Move the Instrument on page 95 to move the transport slide and the plate drawer into the position to accept the transport locks and to remove the cables from the instrument.
  • Page 98 SpectraMax iD3 Multi-Mode Microplate Reader User Guide 5. Route the yellow tab connected to the transport lock so that it will pass over the top of the plate door when the door is closed. 6. Gently close the plate door.
  • Page 99 Chapter 7: Maintenance and Troubleshooting 6. Place the instrument and accessories tool box on the flat cardboard piece and slide it into the original box. 7. Fold up the cardboard flap inside the box. 8. Along the side labeled Open Here, close the box and seal it with packing tape. 5054747 H...

  • Page 100: Obtaining Support

    Knowledge Base, you can submit a request to Molecular Devices Technical Support. You can contact your local representative or Molecular Devices Technical Support at 800-635-5577 x 1815 (North America only) or +1 408-747-1700. In Europe call +44 ...

  • Page 101: Appendix A: Instrument Specifications

    Appendix A: Instrument Specifications This appendix provides specifications for the iD3 basic instrument. Measurement Specifications The following tables list the instrument measurement specifications. Table A-1: Read Times Using Quick Read (plate in/out may add 10-15 seconds) Read Mode 96-Wells 384-Wells Absorbance 0.5 min 1.5 min...
  • Page 102 SpectraMax iD3 Multi-Mode Microplate Reader User Guide Table A-3: Fluorescence Intensity Measurement Specifications Item Description Wavelength range EX 250 - 830 nm EM 270 - 850 nm Wavelength Monochromators tunable in 1.0 nm increments selection Wavelength ±2 nm accuracy Wavelength ±1 nm precision Bandwidth (EX/EM) EX: 15 nm...
  • Page 103 Appendix A: Instrument Specifications Table A-4: Luminescence Measurement Specifications Item Description Wavelength Choice of simultaneous detection of All Wavelengths or selection in 1.0 nm increments selection Wavelength 300 - 850 nm range 300 - 650 nm for "All Wavelengths" setting Wavelength ±2 nm accuracy Wavelength ±1 nm...

  • Page 104: Physical Specifications

    Maximum height: 22 mm Reading Plates and cuvettes (with adapter) SpectraCuvette Adapters without a sticker have a capability plate height of 24 mm and cannot be used in the SpectraMax iD3 or SpectraMax iD5. Robotic compatible Shake Orbital, double orbital, and linear Temperature 5°C (7.2°...
  • Page 105 Appendix A: Instrument Specifications Table A-5: Physical Specifications (continued) Item Description Ambient storage -5°C to 40°C (23°F to 104°F) continuous; temperature -20°C to 50°C (-4°F to 122°F) transient (up to 10 hours) Humidity 15% to 75% (non-condensing) at 30°C (86°F) restrictions Altitude Up to 2000 m (6562 ft)

  • Page 106: Injector Specifications

    SpectraMax iD3 Multi-Mode Microplate Reader User Guide Injector Specifications When your instrument has the SpectraMax Injector System, the specifications for measurements using the injector are shown in the following table. Table A-6: Measurement Specifications For the Injector Item Description Name...
  • Page 107 Appendix A: Instrument Specifications Table A-7: Plate Selection Guidelines For the Injector Read Mode Plate Type Other Considerations Luminescence Solid white When an application specifies a surface (LUM), top read If luminescence crosstalk is high, treatment, use only plates with the correct then use a black plate to improve treatment.
  • Page 108 SpectraMax iD3 Multi-Mode Microplate Reader User Guide 5054747 H...

  • Page 109: Appendix B: Accessories

    7200 Validation Plate iD5*, SpectraMax Paradigm * Specific read modes or cartridges. Injector Accessories The injector and the following accessories are available to order from Molecular Devices. Injector and Injector Accessories Part Number Description ID5-INJ-UPG Injector upgrade kit (Installed by Molecular Devices)
  • Page 110 For a complete substance compatibility list, visit the knowledge base on the Molecular Devices technical support site. Before you use a substance that is not listed, contact Molecular Devices technical support. See Obtaining Support on page 100.
  • Page 111 The trademarks used herein are the property of Molecular Devices, LLC or their respective owners. ©2022 Molecular Devices, LLC. Specifications subject to change without notice. Patents: www.moleculardevices.com/patents All rights reserved. FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES...

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