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 B
May 2017

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  • Page 1 SpectraMax® iD3 Multi-Mode Microplate Reader User Guide 5054747 B May 2017...
  • 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 Before You Operate the Instrument Chapter 1: Introduction Computer Integration Applications Optics Microplate Controls Temperature Regulation Injectors Chapter 2: Unpack and Setup the Instrument Contents of the Package Unpack the Instrument Remove Transport Locks Remove Transport Locks - Alternate Connect Instrument Cables Turn the Instrument On and Off...

  • Page 4: Table Of Contents

    SpectraMax iD3 Multi-Mode Microplate Reader User Guide Define Protocol Settings View Protocol Results Chapter 5: Read Modes and Read Types Endpoint Read Type Kinetic Read Type Spectrum Read Type Absorbance Read Mode Fluorescence Intensity Read Mode Luminescence Read Mode Chapter 6: Maintenance and Troubleshooting...

  • Page 5: Safety Information

    When warnings and cautions appear in this guide, be careful to follow the specific safety information related to them. ® The following user-attention statements can display in the text of Molecular Devices user documentation. Each statement implies a particular amount of observation or...
  • 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. The following table lists the alert symbols on the instrument. Table 3-1: Instrument Label Alert Symbols...
  • Page 7 Safety Information The following sticker appears on the back of your instrument. Table 3-2: Instrument Label Symbols Symbol Indication This symbol indicates the location of a fuse. This symbol indicates that you must consult the product documentation. This symbol indicates the instrument serial number. This symbol indicates the instrument manufacture date.

  • Page 8: Before You Operate The Instrument

    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 9 Safety Information Do not remove the instrument covers. To prevent electrical shock, use the supplied power cords only and connect to a properly grounded wall outlet. Use only multi-plug power strips that are provided by the manufacturer. To ensure sufficient ventilation and provide access for disconnecting power from the instrument, maintain a 20 cm to 30 cm (7.9 in.
  • Page 10 SpectraMax iD3 Multi-Mode Microplate Reader User Guide CAUTION! When you use aggressive or corrosive reagents, you should turn the Auto Eject Plate setting on the Maintenance page Reader Settings tab to the On position. Reader Settings Maintenance on page WARNING! BIOHAZARD.

  • 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 for statistical data analysis. The instrument supports the following read modes:...

  • Page 12: Applications

    GxP mode. You cannot use the Instrument Connection dialog to disconnect from an SpectraMax iD3 instrument that is locked in GxP mode. The SpectraMax iD3 software version 1.0 is compatible with SoftMax Pro Software version 7.0.1. The SpectraMax iD3 software version 1.1 is compatible with SoftMax Pro Software version 7.0.2.

  • Page 13: Microplate Controls

    Chapter 1: Introduction CAUTION! Use of organic solvents can cause harm to the optics in the instrument. Extreme caution is recommended when using organic solvents. Always use a plate lid and do not place a plate containing these materials in the microplate chamber for prolonged periods of time.

  • Page 14: Injectors

    SpectraMax iD3 Multi-Mode Microplate Reader User Guide Injectors When your instrument is equipped with the SpectraMax Injector System, the injectors in the instrument enable you to deliver a reagent to the wells of a microplate. You use the injectors to deliver a specific volume of reagent to initiate a reaction that occurs rapidly and results in a luminescent or fluorescent signal that must be detected quickly.
  • Page 15 Chapter 1: Introduction The SoftMax Pro Software can set up the instrument to inject and read well by well to reduce signal loss. To define the settings for a read with injection, you must use the Acquisition View on the Settings dialog. See the SoftMax Pro Software application help for details. The Injector Maintenance page enables you to wash and prime the injectors.
  • Page 16 SpectraMax iD3 Multi-Mode Microplate Reader User Guide 5054747 B...

  • Page 17: Chapter 2: Unpack And Setup The Instrument

    Chapter 2: Unpack and Setup the Instrument Before you unpack and setup the SpectraMax® iD3 Multi-Mode Plate Reader instrument, prepare a dry, flat work area that has sufficient space for the instrument and required cables. Instrument Specifications on page 125 All software required to run basic non-injector reads is installed in the instrument and is accessible from the touchscreen.

  • Page 18: Contents Of The Package

    SpectraMax iD3 Multi-Mode Microplate Reader User Guide Contents of the Package The package contains the instrument plus a box that contains the tools and accessories required to install the instrument. Table 2-1: Package Contents Illustration Part Description Number 5054744 Installation guide YW 000 Hex key, 2.0 mm...
  • Page 19 5044164 Tubing 5055251 Bottle Holder 5044165 Bottle Adapters 5044163 Waste Plate Cannot order from Molecular Devices Bottles Wide-neck bottle, HDPE 50 mL capacity 36 mm square by 68 mm high 24 mm diameter inside neck Recommended supplier: VWR (215-0440) Cannot order from Molecular Devices...

  • Page 20: Unpack The Instrument

    SpectraMax iD3 Multi-Mode Microplate Reader User Guide Unpack the Instrument The packaging is designed to protect the instrument during transportation. Transport locks are placed on the transport slide and the microplate drawer to protect the instrument from damage during shipment. You must remove the transport locks before you power on the instrument.
  • Page 21 Chapter 2: Unpack and Setup the Instrument 3. Grasp the handle on the cardboard and slide the instrument out of the box. Tip: It might be easier if a second person holds the box in place while the instrument is slid out on the cardboard. Figure 2-2: Slide the instrument out of the box 5054747 B...
  • Page 22 SpectraMax iD3 Multi-Mode Microplate Reader User Guide 4. Remove the accessories tool box. Figure 2-3: Remove the accessory tool box and foam packing CAUTION! Keep the instrument upright and level when lifting. Do not tip or shake the instrument to prevent damage to the moving components inside the instrument.

  • Page 23: Remove Transport Locks

    Chapter 2: Unpack and Setup the Instrument Remove 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 microplate drawer to protect the instrument during shipment.
  • Page 24 SpectraMax iD3 Multi-Mode Microplate Reader User Guide Figure 2-4: Transport lock opening 2. On the front of the instrument, gently pull the yellow tab protruding from the microplate chamber door to open the door. You must hold the microplate drawer door open while you remove the transport lock.

  • Page 25: Remove Transport Locks - Alternate

    Chapter 2: Unpack and Setup the Instrument Figure 2-5: Microplate drawer transport lock (continued) Item Description 3  Screw #3 fastens the lock to the microplate drawer 4  Microplate drawer 5  Microplate chamber door in open position 6  Microplate drawer transport lock 4.
  • Page 26 SpectraMax iD3 Multi-Mode Microplate Reader User Guide To remove the transport lock: 1. Use the provided 4.0 mm hex key to loosen screw #1 in the upper-right corner on the back of the instrument. After a few turns, the touchscreen housing rises. You might hear a click when the housing is ready for the next step.
  • Page 27 Chapter 2: Unpack and Setup the Instrument 3. Loosen screw #2, by hand, to free the transport lock face plate. The screw is attached to the faceplate to prevent it from being misplaced. Figure 2-8: Transport lock face plate 4. Remove the transport lock face plate (with screw #2 attached) to access the transport locks.
  • Page 28 SpectraMax iD3 Multi-Mode Microplate Reader User Guide 7. Screw the long transport lock #4 into the slot on the instrument case from where you removed the short transport lock. Figure 2-11: Switch transport locks 8. Move the transport slide toward the back of the instrument and screw the short transport lock #3 into the lower slot from where you removed the long transport lock.

  • Page 29: Connect Instrument Cables

    Chapter 2: Unpack and Setup the Instrument Figure 2-13: Lower touchscreen housing 12. Walk to the back of the instrument. Use the provided 4.0 mm hex key to tighten screw #1 in the upper-right corner on the back of the instrument until the touchscreen housing lowers and is locked in place.
  • Page 30 SpectraMax iD3 Multi-Mode Microplate Reader User Guide You can use the SoftMax Pro Software on a connected computer to operate the instrument and you must use the SoftMax Pro Software to run injector reads. See the SoftMax Pro User Guide. Table 2-5: Required Accessories...

  • Page 31: Turn The Instrument On And Off

    Chapter 2: Unpack and Setup the Instrument You can optionally connect the instrument directly to a computer. 1. Turn on the power to the computer. 2. Connect one end of an Ethernet cable to the port on the rear of the instrument and then connect the other end of the Ethernet cable to the Ethernet port on the computer.

  • Page 32: Get The Instrument On Your Network

    SpectraMax iD3 Multi-Mode Microplate Reader User Guide Figure 2-16: Touchscreen, power button, NFC sensor 1. To turn on the instrument, press the power button below the touchscreen. 2. Wait until the Home page displays and the disappears. See Home Page on page 3.
  • Page 33 Chapter 2: Unpack and Setup the Instrument Figure 2-17: Ethernet port - instrument rear Tip: You might have to work with your IT department to make sure your network will accept the addition of the instrument. CAUTION! You must remove the transport locks before you power on the instrument.
  • Page 34 SpectraMax iD3 Multi-Mode Microplate Reader User Guide Note: If you plan to use a computer running the SoftMax Pro Software to operate the instrument, write down the IP address. You may need the IP address to connect the computer to the instrument.

  • Page 35: Install The Quicksync Tool

    Chapter 2: Unpack and Setup the Instrument Install the QuickSync Tool The QuickSync tool enables a computer to receive the raw data that the instrument exports, within the security and firewall restrictions of your network. When you install the SoftMax Pro Software on a computer, the SoftMax Pro Software installation process automatically installs the QuickSync tool.
  • Page 36 SpectraMax iD3 Multi-Mode Microplate Reader User Guide 9. Right-click in the tray near the clock to display a menu and select Available Services to display the list of SpectraMax iD3 Multi-Mode Microplate Reader instruments on your intranet and/or the instrument to which you connect the computer through an Ethernet cable.

  • Page 37: Injector Assembly And Maintenance

    Chapter 2: Unpack and Setup the Instrument Injector Assembly and Maintenance This topic describes how to assemble the injectors when you receive a new instrument that has the SpectraMax Injector System. The steps in this topic also describe how to replace the bottle holder and the injector tubing.
  • Page 38 SpectraMax iD3 Multi-Mode Microplate Reader User Guide Bottle Holder The bottle holder is mounted on two knobs for ease of installation, cleaning, and replacement. Figure 2-23: Bottle holder To install the bottle holder: 1. Align the two slots on the bottle holder with the two pegs on the rail.
  • Page 39 Chapter 2: Unpack and Setup the Instrument 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 40 SpectraMax iD3 Multi-Mode Microplate Reader User Guide Figure 2-25: Move injector arm and insert nozzle Perform each of the following steps for both injectors. 2. Insert the snorkel into the bottle and then insert the snorkel into the snorkel clamp.
  • Page 41 Chapter 2: Unpack and Setup the Instrument Figure 2-27: Install tubes 4. Slide the tip into the nozzle, the slots are labeled 1 and 2 for injector 1 and injector 2. Figure 2-28: Insert tips in nozzle 5. Use the black pin 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 42 SpectraMax iD3 Multi-Mode Microplate Reader User Guide Figure 2-29: Insert nozzle into injector port 6. Move the injector arm to its original position. 5054747 B...

  • Page 43: Chapter 3: Home

    Chapter 3: Home Page 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 SoftMax Pro Software to operate the instrument for advanced acquisition settings and for protocols that use the SpectraMax Injector System.

  • Page 44: Protocol Libraries

    SpectraMax iD3 Multi-Mode Microplate Reader User Guide The Home page provides the following controls. Create a New Protocol - Touch to create a protocol. The Create New Protocol dialog appears. Standard Library - Touch to select from a list of pre-defined protocols. See...
  • Page 45 Chapter 3: Home Page Figure 3-2: Protocol libraries on the Home page From the Last Used Documents list, touch a protocol to display the protocol settings (see View Protocol Settings on page 69) or touch a result to display the read results (see View Protocol Results on page 82).

  • Page 46: Quick Read

    SpectraMax iD3 Multi-Mode Microplate Reader User Guide 2. To toggle between the two lists, touch either Standard Library or My Protocols above the protocol list. Touch to sort the protocols in either list alphabetically or by date. In the My Protocols list, touch to add a protocol to your My Protocols list.
  • Page 47 Chapter 3: Home Page Figure 3-4: External Shake dialog To shake a microplate outside the instrument, independently from a protocol: 1. From the buttons on the left, touch to open the microplate drawer and insert the microplate to shake. Note: Leave the microplate drawer open. 2.

  • Page 48: Microplate Chamber Temperature Settings

    SpectraMax iD3 Multi-Mode Microplate Reader User Guide 8. Touch one of the following: Touch Resume to start the shake again. Touch Close Drawer to close the microplate drawer to start a read. Touch Remove Plate to remove the plate from the microplate drawer.

  • Page 49: Injector Maintenance - Wash And Prime

    You should wash the injector tubing before you switch the reagents you use for your experiments. Molecular Devices recommends that you wash the injector tubing with deionized water. For a list of compatible solutions, see...
  • Page 50 SpectraMax iD3 Multi-Mode Microplate Reader User Guide From the buttons on the left, touch to display the Preparations tab on the Injector Maintenance page. The Preparation tab instructions have several pages. You do not need to scroll through the instruction pages to perform the wash and prime steps when you are familiar with the wash and prime procedures.
  • Page 51 Chapter 3: Home Page Note: You must use a computer running SoftMax Pro Software to operate the instrument for advanced acquisition or injector protocols. Wash Injector Tubing To ensure optimal operation of the injector, periodically wash the injector tubing. Molecular Devices recommends that you wash the injector tubing with deionized or distilled water for rinse cycles and 70% alcohol for a disinfectant cycle.
  • Page 52 SpectraMax iD3 Multi-Mode Microplate Reader User Guide The Manual settings enable you control the amount of liquid to move through the injector tubing for a wash or prime. 4. Touch to open the microplate drawer. 5. Insert an empty waste plate and empty strip wells in the microplate drawer. See...
  • Page 53 Chapter 3: Home Page to simultaneously run both injectors for the wash. Start Wash to dispense 900 µl for the wash. Manual to use a keypad to define the amount of liquid to dispense then touch Start. 16. After the third wash step completes, empty the bottles and optionally return them to the bottle holder.
  • Page 54 SpectraMax iD3 Multi-Mode Microplate Reader User Guide Figure 3-8: Injector Maintenance - Prime steps The Injector Maintenance page enables you to perform the wash and prime steps for the injectors. 1. To view the Injector Maintenance page, touch The Manual settings enable you control the amount of liquid to move through the injector tubing for a wash or prime.
  • Page 55 Chapter 3: Home Page 10. Touch: Start Prime for injector 1 to dispense 300 µl from bottle #1. If the protocol uses both bottles, touch Start Prime for injector 2 after the first prime operation completes. Manual to use a keypad to define the amount of liquid to dispense then touch Start.
  • Page 56 SpectraMax iD3 Multi-Mode Microplate Reader User Guide Figure 3-9: Waste plate and strip wells Insert the empty waste plate in the same location as a microplate. Insert the empty strip wells in the smaller slot next to the microplate. When you are ready to run an experiment, replace the waste plate with your prepared microplate.
  • Page 57 Chapter 3: Home Page Figure 3-10: Injector arm in position over 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 touch Wash or Prime.
  • Page 58 SpectraMax iD3 Multi-Mode Microplate Reader User Guide Figure 3-11: Injector bottle holders Place bottle #1 on the left and bottle #2 on the right. The tubes from the snorkel holders to the injector pumps should not cross. The injector comes with adapters that you can insert in the bottle holder to accommodate smaller labware.
  • Page 59 Chapter 3: Home Page 3. Twist the snorkel clamp to clear the position where the bottle is to be placed. 4. Remove the old bottle, if present, and then place the new bottle into its position. 5. Move the snorkel clamp back into position over the bottle. 6.
  • Page 60 SpectraMax iD3 Multi-Mode Microplate Reader User Guide Figure 3-14: Injector Maintenance page - Status tab You must use a computer running SoftMax Pro Software to operate the instrument for advanced acquisition or injector protocols. The SoftMax Pro Software displays messages as the dispensed volume reaches the following milestones: After 2000 ml have been dispensed through the tubing, a message appears to remind...

  • Page 61: Maintenance

    Chapter 3: Home Page Maintenance Page The Maintenance page enables you to manage users, view instrument information, manage reader settings, manage system settings, and to do support tasks such as view instructional videos, export support log files, and to set the transport slide in a position to accept the physical transport lock for shipment and storage.
  • Page 62 SpectraMax iD3 Multi-Mode Microplate Reader User Guide Users Maintenance The Users tab appears selected by default when you first access the Maintenance page. Figure 3-16: Maintenance page - Users tab The first time you power on the instrument, there is only one user - Public. You cannot edit or delete the Public user and Public has no associated NFC tag.
  • Page 63 Chapter 3: Home Page Instrument Maintenance The Instrument tab on the Maintenance page enables you to view instrument information. Figure 3-17: Maintenance page - Instrument tab If you power off the instrument or if you disconnect the Ethernet cable and then reconnect the Ethernet cable, the instrument IP address can change.
  • Page 64 SpectraMax iD3 Multi-Mode Microplate Reader User Guide Reader Settings Maintenance The Reader Settings tab on the Maintenance page enables you to define preferences for protocol reads. Figure 3-18: Maintenance page - Reader Settings tab From the buttons on the left, touch to display the Maintenance page.
  • Page 65 Chapter 3: Home Page System Settings Maintenance The System Settings tab on the Maintenance page enables you to adjust instrument settings. Figure 3-19: Maintenance page - System Settings tab From the buttons on the left, touch to display the Maintenance page. Touch System Settings to display the System Settings tab.
  • Page 66 SpectraMax iD3 Multi-Mode Microplate Reader User Guide Support The Support tab on the Maintenance page enables you to view how-to videos, to export log files to a location from where you can send the log file to a support engineer, and to move the transport slide into a position where it can accept the physical transport lock in preparation for instrument shipment or storage.

  • Page 67: Change User

    Chapter 3: Home Page Change User The Change a User page enables you to define which protocols display in the Last Used Documents section and in the My Protocols list. See Protocol Libraries on page If you use NFC tags, you swipe your NFC tag over the sensor below the touchscreen to change users.

  • Page 68: Load And Unload Microplates

    SpectraMax iD3 Multi-Mode Microplate Reader User Guide Load and Unload Microplates The buttons on the left include to enable you to load or unload a microplate. 1. Touch to move the microplate drawer outside of the instrument. 2. Place the microplate onto the drawer or remove the microplate from the drawer.

  • Page 69: Chapter 4: Protocols

    For instruction about how to use the instrument to define protocol-specific settings, see View Protocol Settings on page 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. View Protocol Settings You can use a quick read protocol, create a protocol from scratch, or you can copy an existing protocol and save it with a new name.
  • Page 70 SpectraMax iD3 Multi-Mode Microplate Reader User Guide Figure 4-1: Plate-specific protocol information To view and manage protocol settings: 1. On the Home page, touch Create a New Protocol or select an existing protocol from the Standard Library, My Protocols, or the list of Last Used Documents.

  • Page 71: Define Protocol Settings

    Chapter 4: Protocols appears. Touch to delete the plate you select. 4. Settings for the plate you select appear on the right. Touch to edit the plate-specific protocol settings. See Define Protocol Settings on page 5. Touch Read to read the microplate. 6.
  • Page 72 SpectraMax iD3 Multi-Mode Microplate Reader User Guide 3. The plate name appears at the top center of the page. Touch <plate name> Plate Format/Type Settings to display the Plate Settings page where you can select the plate format and plate type. See Set Plate Format and Plate Type on page 4.
  • Page 73 Chapter 4: Protocols CAUTION! To prevent damage to the instrument, the height of the microplate must not exceed 22 mm, including the lid if the microplate is lidded. The Plate Settings dialog enables you to select the plate format and to select the plate type. Changes you make here affect the other protocol settings.
  • Page 74 SpectraMax iD3 Multi-Mode Microplate Reader User Guide Set Read Mode and Read Type The Read Mode/Type dialog enables you to set the protocol read mode and protocol 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 protocol settings page.
  • Page 75 Chapter 4: Protocols Note: The icon changes depending upon the read mode and read type. This example is for an Absorbance read mode, Endpoint read type. 2. Touch a read mode. - Absorbance - Fluorescence - Luminescence 3. Touch a read type. - Endpoint - Kinetic - Spectrum...
  • Page 76 SpectraMax iD3 Multi-Mode Microplate Reader User Guide You can choose to read an entire microplate or a subset of wells. Columns do not need to start with column one. Wells in the read area that display a shaded background and the well number will be read.
  • Page 77 Chapter 4: Protocols Interlaced Reading Explained When the instrument reads a sample with a small signal, an interference can occur from the afterglow of a very strong emitting adjacent sample that was measured just before. Such cross talk can occur through the wall of a white 384 well microplate. To prevent such interference, select the Interlaced Reading option to read every other well in a checkerboard pattern in the first run and then read the previously omitted wells in a second run.
  • Page 78 SpectraMax iD3 Multi-Mode Microplate Reader User Guide 4. For Absorbance mode Endpoint type reads, touch the Path Check to display to use PathCheck® technology. The temperature-independent PathCheck Pathlength Measurement Technology normalizes your absorbance values to a 1 cm path length based on the near-infrared absorbance of water. See...
  • Page 79 Chapter 4: Protocols 7. Use the keypad to enter the wavelength value then touch outside the keypad to save your setting. Luminescence Mode Wavelength Settings For the Luminescence read mode with the Endpoint and Kinetic read types, the following wavelength settings are available. Touch the All Wavelength to display and use all wavelengths.
  • Page 80 SpectraMax iD3 Multi-Mode Microplate Reader User Guide Precise - The instrument stops above each selected well and does the read. This provides more precise results than the Fast mode for demanding assays. Fast - The instrument continually moves the plate and the read is timed to occur when the plate reaches the read position.
  • Page 81 Chapter 4: Protocols Luminescence Mode Detection Settings For the Luminescence read mode, the following detection settings are available. 1. Touch the Read Height to display a numeric keypad. 2. Use the keypad to enter the read height from the plate then touch anywhere outside the keypad to save your setting.

  • Page 82: View Protocol Results

    SpectraMax iD3 Multi-Mode Microplate Reader User Guide Set Timing For Kinetic read types, the plate-specific settings page enables you to define timing settings. All read mode Kinetic read types have a Timing tab on the left. See Define Protocol Settings...
  • Page 83 Chapter 4: Protocols Touch to manage your results list. Select a result in the list and touch to make a copy of the result. Select one or more results in the list and touch to export the protocol to a USB or to a computer over your intranet.
  • Page 84 SpectraMax iD3 Multi-Mode Microplate Reader User Guide Manage Results From the Result List page or from your Last Used Documents list, touch a result to display the raw data results of a read. The protocol read results appear as follows.
  • Page 85 Chapter 4: Protocols Figure 4-12: Protocol results - heat map Compare Wells For results that enable you to compare data in the wells, do the following to compare data. Figure 4-13: Compare data in multiple wells 1. Touch to compare the results in several wells. The icon spins and an additional icon appears.
  • Page 86 SpectraMax iD3 Multi-Mode Microplate Reader User Guide Well Specific Linear Results Touch 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 87 Chapter 4: Protocols Export Protocol Data If you operate the instrument from a computer running the SoftMax Pro Software, the touchscreen is disabled and all results appear in the SoftMax Pro Software. You do not need to connect the instrument to your network or directly to a computer to export raw result data.
  • Page 88 SpectraMax iD3 Multi-Mode Microplate Reader User Guide 1. From the buttons on the left, touch to display the Maintenance page. 2. Touch the Reader Settings tab to view the Reader Settings tab. 3. Touch the Export Excel (.xls) to display to export read results to an Excel format for further analysis.

  • Page 89: Chapter 5: Read Modes And 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. For information on the supported read modes, see the following topics:...

  • Page 90: Spectrum Read Type

    The reported OD values for turbid samples are likely to be different when read by different instruments. For optimal results, Molecular Devices recommends that you run replicates for all blanks, controls, and samples. In this case, the blank value that can be subtracted is the average value of all blanks.
  • Page 91 Chapter 5: Read Modes and Read Types where I is the intensity of light after it passes through the sample and I is incident light before it enters the sample. Optical Density and %Transmittance are related by the following formulas: 2–OD %T = 10 OD = 2 – log...
  • Page 92 SpectraMax iD3 Multi-Mode Microplate Reader User Guide The standard pathlength of a 1 cm cuvette is the conventional basis for quantifying the unique absorptivity properties of compounds in solution. Quantitative analysis can be done on the basis of extinction coefficients, without standard curves (for example, NADH-based enzyme assays).
  • Page 93 Chapter 5: Read Modes and Read Types Eliminate the Pathlength Independent Component Raw OD measurements of microplate samples include both pathlength-dependent components (sample and solvent) and a pathlength-independent component (OD of microplate material). The pathlength-independent component must be eliminated from the calculation to get valid results that have been normalized by the PathCheck technology.

  • Page 94: Fluorescence Intensity Read Mode

    SpectraMax iD3 Multi-Mode Microplate Reader User Guide Samples that are highly colored in the upper-visible spectrum might have absorbance that extends into the near-infrared (NIR) spectrum and can interfere with the PathCheck technology. Examples include Lowry assays, molybdate-based assays, and samples that contain hemoglobins or porphyrins.
  • Page 95 Chapter 5: Read Modes and Read Types Excitation maximum Emission maximum Stokes Shift Absorption Wavelength (nm) Figure 5-2: 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, you must have sufficient spectral separation to reduce the interference of the excitation light with detection of the emitted light.
  • Page 96 SpectraMax iD3 Multi-Mode Microplate Reader User Guide Excitation maximum of fluorophore Emission maximum of fluorophore Excitation reading wavelength Emission reading wavelength Wavelength (nm) Figure 5-3: Optimized Excitation and Emission Reading Wavelengths The previous figure shows that the best results are often obtained when the excitation and emission wavelengths used for reading are not the same as the peak wavelengths of the excitation and emission spectra of the fluorophore.
  • Page 97 Chapter 5: 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 of this mode is to study the kinetics of ion release. Some assays use a fluorescent label to selectively attach to certain compounds.

  • Page 98: Luminescence Read Mode

    SpectraMax iD3 Multi-Mode Microplate Reader User Guide For optimal results, Molecular Devices recommends that you run replicates for all blanks, controls, and samples. In this case, the blank value that can be subtracted is the average value of all blanks.
  • Page 99 To help eliminate background luminescence from a microplate that has been exposed to light, Molecular Devices recommends dark adaptation of the microplate by placing the sample-loaded microplate in the instrument for several minutes before starting the read.
  • Page 100 For optimal results, Molecular Devices recommends that you run replicates for all blanks, controls, and samples. In this case, the blank value that can be subtracted is the average value of all blanks.
  • Page 101 Chapter 5: Read Modes and Read Types Sample Volumes and Concentration of Reactants The concentration of the luminescent agent impacts the quantity of light output in a luminescent reaction. Light is emitted as a result of a reaction between two or more compounds.
  • Page 102 SpectraMax iD3 Multi-Mode Microplate Reader User Guide 5054747 B...

  • Page 103: Chapter 6: Maintenance And Troubleshooting

    Chapter 6: Maintenance and Troubleshooting 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 Obtain Support on page 124. Before operating the instrument or doing maintenance operations, make sure that you are familiar with the safety information in this guide.

  • Page 104: Preventive Maintenance

    Respond as required to all error messages displayed by the software. Molecular Devices recommends turning the power off when the instrument is not in use. Replace Fuses If the instrument does not seem to get power after you switch it on, check to see whether the power cord is securely plugged into a functioning power outlet and to the power port on the rear of the instrument.
  • Page 105 7. Plug the power cord into the power port. 8. Turn on power to the instrument. Note: If the instrument still does not power on after you change the fuses, contact Molecular Devices technical support. See Obtain Support on page 124.

  • Page 106: Clean The Instrument

    SpectraMax iD3 Multi-Mode Microplate Reader User Guide Clean the Instrument WARNING! BIOHAZARD. Always wear gloves when operating the instrument and during cleaning procedures that could involve contact with either hazardous or biohazardous materials or fluids. Always turn the power off and disconnect the power cord from the main power source before you use liquids to clean the instrument.
  • Page 107 Chapter 6: Maintenance and Troubleshooting Figure 6-2: Maintenance page - Support tab - Lock Instrument button Before you power off the instrument, do the following to prepare the instrument for a move. 1. Touch and remove the microplate from the microplate drawer, if present. 2.
  • Page 108 SpectraMax iD3 Multi-Mode Microplate Reader User Guide Note: The screw remains inside the instrument. The screw is spring mounted and cannot get lost within the instrument. Tighten the screw until it is snug. This locks the transport slide. 7. Move to the front of the instrument and touch OK to confirm that the transport slide has been locked.
  • Page 109 Chapter 6: Maintenance and Troubleshooting Table 6-2: Required Tool Illustration Part Number Description YW 000 006 Hex key, 2.0 mm YW 000 012 Holex HEXAGON ballhead bolt driver 3 mm When you do all the steps in Before You Move the Instrument on page 106 section, the transport slide moves to the correct position, you lock the transport slide, the microplate drawer door opens, and the microplate drawer moves into the position to accept the...
  • Page 110 SpectraMax iD3 Multi-Mode Microplate Reader User Guide 3. Gently push the microplate drawer into the instrument and as far to the left as possible until screw #1, which fastens the lock to the internal frame of the instrument, lines up with the hole on the internal frame.
  • Page 111 Chapter 6: Maintenance and Troubleshooting touchscreen is unlocked. Figure 6-5: Instrument rear - release touchscreen housing 2. Walk to the front of the instrument and gently lift the touchscreen housing to access the transport lock. Figure 6-6: Instrument front - raise touchscreen housing WARNING! You must hold the touchscreen housing in place for the following steps.
  • Page 112 SpectraMax iD3 Multi-Mode Microplate Reader User Guide 4. Remove the transport lock face plate (with screw #2 attached) to access the transport locks. Figure 6-8: Remove transport lock face plate 5. By hand, remove the long transport lock #4 from the slot on the instrument case.
  • Page 113 Chapter 6: Maintenance and Troubleshooting 9. Align the transport lock faceplate with screw #2 to its original position. Figure 6-11: Replace transport lock face plate 10. Screw the transport lock faceplate back into place with screw #2 by hand. 11. Gently lower the touchscreen housing. Note: Do not push the touchscreen housing down manually.
  • Page 114 SpectraMax iD3 Multi-Mode Microplate Reader User Guide 12. Walk to the back of the instrument. Use the provided 4.0 mm hex key to tighten screw #1 in the upper-right corner on the back of the instrument until the touchscreen housing lowers and is locked in place. Do not over tighten the screw.
  • Page 115 Chapter 6: Maintenance and Troubleshooting 1. Use the provided 4.0 mm hex key to loosen screw #1 in the upper-right corner on the back of the instrument. After a few turns, the touchscreen housing rises. You might hear a click when the housing is free for the next step. Do not over loosen screw after touchscreen housing is unlocked.
  • Page 116 SpectraMax iD3 Multi-Mode Microplate Reader User Guide 3. Loosen screw #2, by hand, to free the transport lock face plate. The screw is attached to the faceplate to prevent it from being misplaced. Figure 6-16: Transport lock face plate 4. Remove the transport lock face plate (with screw #2 attached) to access the transport locks.
  • Page 117 Chapter 6: Maintenance and Troubleshooting Figure 6-19: Switch transport locks Screw the short transport lock #3 into the slot on the instrument case from where you removed the long transport lock. Before you screw the long transport lock into place, perform the following steps to align the transport slide and plate transport drawer.
  • Page 118 SpectraMax iD3 Multi-Mode Microplate Reader User Guide Figure 6-22: Align plate transport with chamber opening 10. Push the plate transport slightly into the transport chamber until the front of the plate transport aligns with the front of the chamber opening.
  • Page 119 Chapter 6: Maintenance and Troubleshooting Figure 6-24: Set plate transport on bolt. ^ Top view ^ 13. Move the plate transport a little bit inside and then to the left until you feel the hole on the front of the plate transport catch the bolt. Do not apply any downward pressure on the transport as you slide the plate over the bolt.
  • Page 120 SpectraMax iD3 Multi-Mode Microplate Reader User Guide 16. Align the transport lock faceplate with screw #2 to its original position. Figure 6-27: Replace transport lock face plate 17. Screw the transport lock faceplate back into place with screw #2 by hand.
  • Page 121 Chapter 6: Maintenance and Troubleshooting 19. Walk to the back of the instrument. Use the provided 4.0 mm hex key to tighten screw #1 in the upper-right corner on the back of the instrument until the touchscreen housing lowers and is locked in place. Do not over tighten screw. Figure 6-29: Lock touchscreen housing For steps to pack the instrument in its original container, see Pack the Instrument on page...
  • Page 122 SpectraMax iD3 Multi-Mode Microplate Reader User Guide 4. Replace the molded foam packaging around the instrument. Figure 6-30: Accessory box and foam packaging CAUTION! Keep the instrument upright and level when lifting. Do not tip or shake the instrument to prevent damage to the moving components inside the instrument.

  • Page 123: 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 Obtain Support on page 124.

  • Page 124: Obtain Support

    Technical Support Service Request Form to send an email to our technical support representatives. You can contact your local representative or contact Molecular Devices Technical Support by telephone at 800-635-5577 (North America only) or +1 408-747-1700. In Europe call +44 (0) 118 944 8000.

  • Page 125: 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) 96 Wells 384 Wells Absorbance 0.5 min 1.5 min...
  • Page 126 SpectraMax iD3 Multi-Mode Microplate Reader User Guide Table A-3: Fluorescence Intensity Measurement Specification of the iD3 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 127 Appendix A: Instrument Specifications Table A-4: Luminescence Measurement Specifications of the iD3 Item Description Wavelength Choice of simultaneous detection of All Wavelengths or selection in 1.0 nm Selection increments Wavelength 300 - 850 nm Range 300 - 650 nm for "All Wavelengths" setting Wavelength ±2 nm Accuracy...

  • Page 128: Computer System Specifications

    You can set these options in Windows Control Panel. The SpectraMax iD3 software version 1.0 is compatible with SoftMax Pro Software version 7.0.1. The SpectraMax iD3 software version 1.1 is compatible with SoftMax Pro Software version 7.0.2.

  • Page 129: Physical Specifications

    Appendix A: Instrument Specifications Physical Specifications The following tables list the physical specifications of the SpectraMax iD3 instrument. Table A-6: Physical Specifications of the iD3 Instrument Item Description Environment Indoor Use Only Power 100-240 VAC ±10%, 2 A, 50/60 Hz...

  • Page 130: Injector Specifications

    SpectraMax iD3 Multi-Mode Microplate Reader User Guide Table A-6: Physical Specifications of the iD3 Instrument (continued) Item Description Altitude Up to 2000 m (6562 ft) Restrictions Air Pressure 54 kPa to 106 kPa (7.8 PSI to 15.4 PSI) Restrictions Sound Pressure Maximum sound pressure: 73 dBA Level Maximum sound pressure at one meter: 68 dBA...
  • Page 131 Appendix A: Instrument Specifications Table A-7: Measurement Specifications for the Injector (continued) Item Description Injectors Dispense volume 1 µL increments from 1 µL to the maximum allowable volume of the well, based on the selected microplate type Dispense accuracy ±(4% of volume + 1 μL) / volume x 100% Dispense precision ≤(2% of volume + 1 μL) / volume [μL] x 100% cv Dispense speed...
  • Page 132 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. Obtain Support on page 124.

  • Page 133: Appendix A: System Diagrams And Dimensions

    Appendix A: System Diagrams and Dimensions Table A-10: List of Compatible and Incompatible Solutions Compatible Solutions Do Not Use Alcohol, Ethyl (Ethanol), 70% solution or less Acetone Alcohol, Isobutyl (Isobutanol), 70% solution or less Alcohol, Benzyl (Phenylcarbinol) Alcohol, Methyl (Methanol), 70% solution or less Hydrochloric Acid (HCl) Ammonia, 10% solution or less Ketones...
  • Page 134 SpectraMax iD3 Multi-Mode Microplate Reader User Guide Figure A-1: Front View of the iD3 with Dimensions (continued) Item Description 3  Height of Microplate Drawer: 9.5 cm (3.7 in.) 5054747 B...
  • Page 135 Appendix A: System Diagrams and Dimensions Figure A-2: Side View of the iD3 with Dimensions Item Description 1  Depth: 59.8 cm (23.54 in.) 2  Height: 40.1 cm (15.79 in.) 3  Maximum Length of Opened Microplate Drawer: 11 cm (4.33 in.) 5054747 B...
  • Page 136 SpectraMax iD3 Multi-Mode Microplate Reader User Guide 5054747 B...

  • Page 137: Appendix B: Electromagnetic Compatibility

    Appendix B: Electromagnetic Compatibility Regulatory for Canada (ICES/NMB-001:2006) This ISM device complies with Canadian ICES-001. Cet appareil ISM est confomre à la norme NMB-001 du Canada. ISM Equipment Classification (Group 1, Class A) This equipment is designated as scientific equipment for laboratory use that intentionally generate and/or use conductively coupled radio-frequency energy for internal functioning, and are suitable for use in all establishments, other than domestic and those directly connected to a low voltage power supply network which supply buildings used for domestic...
  • Page 138 SpectraMax iD3 Multi-Mode Microplate Reader User Guide 5054747 B...

  • Page 139: Index

    Index fluorescence intensity FL 94 read mode 94 fuses absorbance 90 replacing 104 read mode 90 absorbance ABS 90 read mode 90 hardware requirements 128 height detection of microplates 13 cleaning instrument 106 instrument safety 123 preventive maintenance 104 computer requirements 128 replacing fuses 104 customer support 124 luminescence 99...
  • Page 140 SpectraMax iD3 Multi-Mode Microplate Reader User Guide water constant 92 microplate height detection 13 specifications loading and unloading 68 instrument dimensions 133 instrument measurements 125 moving parts 10 physical 129 spectrum read type 90 organic solvents 9, 13 system requirements 128...
  • Page 141 The trademarks used herein are the property of Molecular Devices, LLC or their respective owners. ©2017 Molecular Devices, LLC. Specifications subject to change without notice. Patents: www.moleculardevices.com/productpatents All rights reserved. FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES...

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