Sony ID7000 Spectral Cell Analyser
Designed to Empower Deep Scientific Insights
The ID7000™ spectral cell analyser supports high-parameter flow cytometry by delivering comprehensive information about heterogeneous cell populations, with high sensitivity to detect dim and rare populations.
Expanding the capabilities of cell analysis, the ID7000 can be configured with up to 7 lasers and 186 detectors, the most of any flow cytometer available today. The system enables researchers to perform experiments using 44 colors or more, limited only by the fluorochromes available. Laser choices range from deep ultraviolet (320 nm) to infrared (808 nm). The system’s detection capability enables expansion of multicolor panels and can keep pace with future fluorochrome development.
| Designed for Multicolour Workflows
|
The advanced optics design, along with other proprietary technologies included in the ID7000 spectral system, provide best-in-class data so that researchers can collect the most information from every sample. Sony has developed signal processing technologies to reduce electronic noise, central to achieving high sensitivity in a flow cytometer. Together, these capabilities help to deliver the sensitivity to help scientists achieve more accurate results. The new excitation and detection optics in the ID7000 allow users to select spectrally similar fluorochromes and resolve the signal with confidence, to gain a deeper understanding of cellular biology. As pioneers in spectral technology, we at Sony have designed the optics to unlock the full potential of spectral analysis, with an unparalleled number of lasers and PMT detectors that provide the resolution needed to unmix even the most similar spectral signatures. |
| Excitation optics | The excitation optics are designed to maximise fluorochrome excitation and emission across the detector array to deliver distinctly resolved populations. The system is equipped with three standard excitation lasers in all configurations: the 488-nm (blue), 637-nm (red), and 405-nm (violet) lasers. Optional 561-nm (yellow-green), 355-nm (ultraviolet), 320-nm (deep ultraviolet), or 808-nm (infrared) lasers support a wide variety of applications. Each excitation laser has power specifications optimised to deliver the highest performance for detecting dim and rare markers. |
| Detector Capabilities | The expanded detector availability addresses the complexity of panel design, and in turn fluorochrome choices, which empowers discovery. Photomultiplier tubes (PMTs) have been chosen for their high signal-to-noise performance to capture the most photons, leading to higher resolution and lower background, enhancing dim signal detection for superior visualisation of rare populations. Emitted light is collected from 360 nm to 920 nm using PMTs (32-channel, single-channel, and single-channel IR-sensitive), and a spectral fingerprint is generated. A light diffraction grating based collection system maximises photon capture across the entire spectrum by distributing light evenly across the surface of each of the 32-channel PMT arrays. This new, robust system provides high resolution across the entire detector array for a larger, more evenly distributed detection range, and minimises light loss. The detection capacity of the ID7000 spectral cell analyser spans from 360 nm to 920 nm in the 7-laser configuration. The signal is captured using a combination of 32-channel PMT arrays and individual PMTs. The signal above 800 nm is detected with specialised InGaAs PMTs to ensure data quality.
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| Small Particle Detection | The ID7000 can detect small particles. The figure shows how a mix of fluorescent beads of varied diameters available as a commercial product (Megamix, Biocytex), is analysed to clearly resolve the 160-nm beads from all other beads and separate from noise, using SSC and channel 7 of the 32-channel PMT array on the blue laser deck.
The cabinets incorporate a built-in aerosol management system which operates independently to actively evacuate aerosols from the sort collection chamber. The dual routes of aerosol evacuation maximise protection. |
| Designed to Harness the Power of Spectral Data | The ID7000 spectral cell analyser builds on Sony’s experience with spectral analysis and simplifies many operations, even for complex experiments. With the true signal for each fluorochrome unaffected by autofluorescence or subjective adjustment of spillover, spectral analysis yields cleaner, unbiased data for every experiment. |
| Spectral Unmixing | In spectral analysis, signals from all detected channels are used to create one spectral emission signal, regardless of the number of fluorochromes analysed. Unmixing, a powerful capability, then separates fluorophores into pure signals that measure the intensity of each fluorophore at each wavelength to more accurately measure data for analysis. The ID7000 uses the WLSM (Weighted Least Squares Method) fluorescence unmixing algorithm to separate the individual spectral fingerprints and enable scientists to analyse dim and rare phenotypic marker expression. Unmixing delivers a more comprehensive picture of rare populations while decreasing the complexities associated with working with fluorescent proteins and fluorochromes excited by multiple lasers. Overall, spectral technology simplifies multicolor panel design by eliminating the use of bandpass filters and highly subjective, conventional compensation matrixes. |
| Autofluorescence Subtraction | Spectral analysis, while allowing researchers to see the full emission signal without using bandpass filters, also enables autofluorescence to be handled as a separate color. In conventional flow cytometry, cellular autofluorescence produced by pyridine (NAD/NADH), flavin (FMN, FAD), and other intracellular oxidative reactions can interfere with signals of other fluorescent markers. Other common sources of autofluorescence include cell fixation and permeabilisation. Spectral technology subtracts one or more autofluorescent spectral fingerprints to allow researchers to see the true fluorescent populations. |
| Reusable Data from a Spectral Library | Users acquire single-color control data before experimental samples so that the unmixing algorithm has emission spectra inputs that it can reference to unmix the experimental sample. Single-color control spectra are stored in the Spectral Reference Library. Unlike compensation controls, spectral references do not need to be acquired for every new experiment and can be reused. This allows users to create a personal or shared reagent library that simplifies experiment creation, saves time and valuable reagents, and increases efficiency. |
| System Standardisation for Reproducible Data | The ID7000 system can be operated in a Normal or Standardisation mode. Standardisation mode sets the system to an optimised master specification that allows researchers to maintain instrument settings between experiments and across multiple instruments to support longitudinal studies and cross-site collaboration. Standardisation mode adjusts the output of each channel so that the SSC and fluorescence detection sensitivities are the same between multiple instruments. PMT voltage correlation coefficients are calculated for each laser detection deck during daily QC, resulting in standardised measurements across all lasers, detectors, and instruments. Standardisation mode voltages can be adjusted synchronously or for each laser detection deck. The system supports standardisation of Area and Height data. This unique capability allows scientists to obtain the best data at maximum PMT voltages and reduces the need to rerun single-color controls for every experiment. Standardisation mode minimises subjectivity and instrument variability to yield highly reliable, accurate, and reproducible results. |
| Designed for Automated Sample Acquisition | Automation is present across the workflow to simplify operation and ensure accurate results. To give scientists more efficiency in their day-to-day work, the ID7000 comes with an advanced, best-in-class AutoSampler, which supports 5-mL tube racks (24 tubes), 96-well standard height, 96-well half deep, and 96-well deep plates, and 384-well standard flat bottom plates. Samples can be cooled to 4°C (39°F) using a Peltier element to prevent signal degradation and reduce variability. The sample in the individual tubes or wells can be agitated at the beginning of acquisition, intermittently or continuously, enabling stable, long-duration acquisition without sample settling. To ensure flexibility, the AutoSampler supports multiple wash options. A Dual Probe Wash option incorporates a cleansing mechanism that moves up and down to clean the inside and outside of the sample nozzle. Other wash options are available when speed of acquisition is the priority. The integration of intelligent software tools allows for management of bubbles and clogs, and integrated sensing mechanisms enable acquisition of critical samples at low volumes. The AutoSampler is equipped with an extra loading station for added convenience. Up to three 5-mL tubes can be placed in the extra positions and used, for example, for loading QC particles to streamline system QC. Or, the wash and cleaning solutions can be loaded and the software programmed for the system to shut down automatically after acquisition is completed. This is invaluable for users performing multi-sample acquisitions that are time consuming. |
| Loading Options | The ID7000 standard AutoSampler supports multiple loading options including a variety of 96-well and 384-well plates, as well as a 24-position tube rack for 12 x 75-mm (5-mL) tubes. |
| AutoSampler Agitation Modes | The ID7000 sample agitation feature is designed to keep large particles in suspension. The figure shows data with 30-µm particles when applying the initial mixing mode (Once) and the Cyclic mixing mode (Cyclic). It shows that the particles remain in suspension for 10 minutes when using the initial mixing mode, before there is an observed reduction in event rate. The Cyclic mixing mode, when used, continually mixes the sample throughout the entire acquisition, resulting in a consistent event rate with no settling. |
| 5-mL Tube Rack Sample Cooling Over Time | The graph above shows how sample temperature is maintained in the ID7000 AutoSampler when using a 24-position tube rack. Temperature measurement for tubes at multiple positions indicate that they maintain cooling over time when used in a 23°C ambient environment. |
| AutoSampler Automated Workflows | The ID7000 AutoSampler includes an extra tube station that accommodates up to three 12 x 75-mm (5-mL) tubes that can be used to support automated QC, system cleaning, and automatic shutdown workflows. |
| Designed for True Ease of Use | The ID7000 system software is easy to learn and use. It guides researchers from setup and QC to panel design, data acquisition, data analysis, through system shutdown. For accommodating users with varied experience levels, most workflows are designed with an expert mode and a guided mode. In guided mode, software wizards simplify system operation. The software provides access levels and tools to suit the needs of busy laboratories. Preferences allow users and administrators to specify options for overall instrument operation and experimental setup to facilitate use of the ID7000 spectral cell analyser. It is also possible to configure software preference settings for each individual user for added convenience. |
| Intuitive Workflows for Acquisition and Analysis | In the ID7000 system software, users can create experiments from a template, an existing experiment, a single-stained sample, or a multicolor assay to accommodate their preferred approaches. New users can create new experiments with the Experiment Design Wizard. To accommodate multi-sample and multiparameter experiment needs, the ID7000 software provides both a Shared Worksheet and an Individual Worksheet. A Shared Worksheet allows each sample in a group to use the same worksheet and thus makes experiment setup more intuitive.
Instrument settings can be specified for each sample group and configured for lasers and detectors to give researchers the needed flexibility. Altering these settings for a sample in a group changes the settings for all unmeasured samples belonging to the same group. Instrument settings can be exported and imported The acquisition functions are all accessible from the Acquisition tab and have been developed keeping a multi-laser workflow in mind. The tab accesses all functions that control the data acquisition, recording, and stopping, as appropriate, while monitoring the data acquisition status. To enable walkaway functionality, the system has an Event Checking function that can be set to stop or skip and continue automatic acquisition if the event rate drops or air bubbles are detected. For sample analysis, the appropriate spectral references must be selected and fluorescence unmixing applied to remove the spillover components from each fluorescence channel. The unmixing can be manually adjusted on the screen using the Spectral Reference Adjuster. Making changes on one plot will result in the changes being applied automatically to all other plots having that fluorochrome as one of their parameters. Users can save the adjustment result as a new fluorescence unmixing matrix or can overwrite an existing fluorescence unmixing matrix. The Analysis tab houses all controls for analysis operations on the ID7000 spectral flow cytometer, providing access to worksheet tools that let users choose how the data is displayed (such as plot types), and customize the display for their analysis needs. Plots and statistics provide real-time information during acquisition. The Experiment Explorer pane allows search by date and keyword, to display desired data on the worksheet. Once displayed, plots can be edited individually or as a group, to facilitate working with complex gating hierarchies. The software also supports batch processes for users who need to run multiple samples, for operations like experiment setup, data acquisition and analysis, and export of data and statistics. Combined with the ability to program an automated shutdown, this enables scientists to focus on discovery instead of repetitive tasks. |
| Automated Setup and Quality Control to Ensure System Performance | At startup, the Align Check and Performance QC wizards use beads to check instrument calibrations, to ensure the instrument is operating optimally. On-screen instructions guide the user through procedures, then display progress and report results. The QC results are available as a report that can be shared and archived. Trends over a desired period of time can also be displayed to ensure that the system performance is consistent as expected. At startup, the Align Check and Performance QC wizards use beads to check instrument calibrations, to ensure the instrument is operating optimally. On-screen instructions guide the user through procedures, then display progress and report results. The QC results are available as a report that can be shared and archived. Trends over a desired period of time can also be displayed to ensure that the system performance is consistent as expected. |
Optics / Performance
| Excitation lasers | 320 nm, 355 nm, 405 nm, 488 nm, 561 nm, 637 nm, 808 nm |
| Detectors | Photomultiplier tubes (32-channel PMTs, single-channel PMTs, and single-channel IR-sensitive PMTs) |
| Fluorescence sensitivity (molecules of equivalent soluble fluorochrome)* | FITC ≤ 58 MESF; PE ≤ 4 MESF; APC ≤ 7 MESF *Specifications listed are an average of data obtained from multiple systems |
| Pulse parameter | Area, Height, Width (all channels) |
| Signal resolution | Height (20 bit), Area (32 bit) Sampling frequency: 60 MHz |
| Detectable particle size | 160 nm with SSC |
| Fluorescence resolution | CV < 3% for the singlet peak of propidium iodide-stained Chicken Erythrocyte Nuclei (CEN) |
| Event Rate | 40,000 events per second |
Fluidics
| Sample flow rate | ~33 μL/min–200 μL/min |
| Cleaning modes | Priming, Probe Wash, Flow Cell Purge, Decontamination (with bleach), Hardware Shutdown (performed before system shutdown), Automatic Shutdown (programmed system shutdown after completion of Auto Acquisition) |
| System fluidics | 10-L capacity Sheath Tank and 10-L Capacity Waste Tank Sheath fluid (distilled water or phosphate buffered saline) |
System Software
| Software | ID7000 Acquisition and Analysis Software |
| Supported file types | exdat and FCS 3.1 |
AutoSampler
| Sample loading options | Multiwell plates: 96-well plate: standard height flat/V/U bottom, 96-well half deep V bottom, 96-well deep U bottom, 384-well standard flat bottom
Tubes: 24-tube rack that accommodates 5-mL (12 x 75-mm) polystyrene/polypropylene tubes |
| Minimum sample volume* | 5-mL tube: 100 µL in standard mode/50 µL in low-volume mode, 96-well plate: 55 µL in standard mode/10 µL in low-volume mode, 384-well plate: 40 µL in standard mode/10 µL in low-volume mode *Minimum volumes provided for the sample vessel type are averages of data obtained from multiple experiments |
| Carryover | < 0.1% (when measured with inner and outer Probe Wash) |
| Throughput | 96-well plate in 19 minutes (Based on an acquisition time of 2 seconds per well, no agitation, and no probe wash. The value reported is an average of multiple experiments.) |
| Sample agitation | Multiple mixing modes: Once at the beginning of acquisition, Cyclic, or Continuous Mixing |
| Temperature Control | From ambient temperature to 4°C with a Peltier sample cooling module |
| Power consumption | 600 W (max.) |
Workstation
| Specifications | Operating System: Windows® 10 Pro 64 bit Processor: Intel® Xeon® 6248 2.5 GHz 20 Core Memory: 256 GB DDR4 Graphics card: NVIDIA Quadro P2200 Storage: 8TB x3 (RAID5:16TB) HDD 2 Ethernet ports 4 display ports DVD disc drive |
Facility requirements
| Dimensions | W: 41.7 in (106 cm) x D: 28.3 in (71.9 cm) x H: 29.9 in (76 cm) |
| Weight | 463 lb (210 kg) |
| Power Consumption | 900 W (2 independent 450-W circuits) |
Regulatory requirements
| Class 1 Laser Product |
| For non-clinical Research Use Only. Not for use in diagnostic or therapeutic procedures or for any other clinical purpose. |
| United States: Class A digital device Canada: CAN ICES-3 (A)/NMB-3(A) EU: EU Declaration of Conformity 2006/42/EC (Machinery); 2014/30/EU (EMC) |
Laser configurations
| Model | No. of lasers, wavelength | No. of detectors* |
|---|---|---|
| LE-ID7000A | 3LD: 405/488/637 | FSC/SSC + 86F |
| LE-ID7000B | 4LD: 405/488/561/637 | FSC/SSC + 112F |
| LE-ID7000C | 5LD: 355/405/488/561/637 | FSC/SSC + 147F |
| LE-ID7000D | 6LD: 355/405/488/561/637/808 | FSC/SSC + 149F |
| LE-ID7000E | 6LD: 320/355/405/488/561/637 | FSC/SSC + 182F |
| LE-ID7000F | 7LD: 320/355/405/488/561/637/808 | FSC/SSC + 184F |
*F = fluorescence
For Research Use Only.
Class 1 Laser Product.
- Features & Benefits
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Designed for Multicolour Workflows The advanced optics design, along with other proprietary technologies included in the ID7000 spectral system, provide best-in-class data so that researchers can collect the most information from every sample. Sony has developed signal processing technologies to reduce electronic noise, central to achieving high sensitivity in a flow cytometer. Together, these capabilities help to deliver the sensitivity to help scientists achieve more accurate results. The new excitation and detection optics in the ID7000 allow users to select spectrally similar fluorochromes and resolve the signal with confidence, to gain a deeper understanding of cellular biology. As pioneers in spectral technology, we at Sony have designed the optics to unlock the full potential of spectral analysis, with an unparalleled number of lasers and PMT detectors that provide the resolution needed to unmix even the most similar spectral signatures. Excitation optics The excitation optics are designed to maximise fluorochrome excitation and emission across the detector array to deliver distinctly resolved populations. The system is equipped with three standard excitation lasers in all configurations: the 488-nm (blue), 637-nm (red), and 405-nm (violet) lasers. Optional 561-nm (yellow-green), 355-nm (ultraviolet), 320-nm (deep ultraviolet), or 808-nm (infrared) lasers support a wide variety of applications. Each excitation laser has power specifications optimised to deliver the highest performance for detecting dim and rare markers. Detector Capabilities The expanded detector availability addresses the complexity of panel design, and in turn fluorochrome choices, which empowers discovery. Photomultiplier tubes (PMTs) have been chosen for their high signal-to-noise performance to capture the most photons, leading to higher resolution and lower background, enhancing dim signal detection for superior visualisation of rare populations. Emitted light is collected from 360 nm to 920 nm using PMTs (32-channel, single-channel, and single-channel IR-sensitive), and a spectral fingerprint is generated. A light diffraction grating based collection system maximises photon capture across the entire spectrum by distributing light evenly across the surface of each of the 32-channel PMT arrays. This new, robust system provides high resolution across the entire detector array for a larger, more evenly distributed detection range, and minimises light loss. The detection capacity of the ID7000 spectral cell analyser spans from 360 nm to 920 nm in the 7-laser configuration. The signal is captured using a combination of 32-channel PMT arrays and individual PMTs. The signal above 800 nm is detected with specialised InGaAs PMTs to ensure data quality. Small Particle Detection The ID7000 can detect small particles. The figure shows how a mix of fluorescent beads of varied diameters available as a commercial product (Megamix, Biocytex), is analysed to clearly resolve the 160-nm beads from all other beads and separate from noise, using SSC and channel 7 of the 32-channel PMT array on the blue laser deck. The cabinets incorporate a built-in aerosol management system which operates independently to actively evacuate aerosols from the sort collection chamber. The dual routes of aerosol evacuation maximise protection.
Designed to Harness the Power of Spectral Data The ID7000 spectral cell analyser builds on Sony’s experience with spectral analysis and simplifies many operations, even for complex experiments. With the true signal for each fluorochrome unaffected by autofluorescence or subjective adjustment of spillover, spectral analysis yields cleaner, unbiased data for every experiment. Spectral Unmixing In spectral analysis, signals from all detected channels are used to create one spectral emission signal, regardless of the number of fluorochromes analysed. Unmixing, a powerful capability, then separates fluorophores into pure signals that measure the intensity of each fluorophore at each wavelength to more accurately measure data for analysis. The ID7000 uses the WLSM (Weighted Least Squares Method) fluorescence unmixing algorithm to separate the individual spectral fingerprints and enable scientists to analyse dim and rare phenotypic marker expression. Unmixing delivers a more comprehensive picture of rare populations while decreasing the complexities associated with working with fluorescent proteins and fluorochromes excited by multiple lasers. Overall, spectral technology simplifies multicolor panel design by eliminating the use of bandpass filters and highly subjective, conventional compensation matrixes. Autofluorescence Subtraction Spectral analysis, while allowing researchers to see the full emission signal without using bandpass filters, also enables autofluorescence to be handled as a separate color. In conventional flow cytometry, cellular autofluorescence produced by pyridine (NAD/NADH), flavin (FMN, FAD), and other intracellular oxidative reactions can interfere with signals of other fluorescent markers. Other common sources of autofluorescence include cell fixation and permeabilisation. Spectral technology subtracts one or more autofluorescent spectral fingerprints to allow researchers to see the true fluorescent populations. Reusable Data from a Spectral Library Users acquire single-color control data before experimental samples so that the unmixing algorithm has emission spectra inputs that it can reference to unmix the experimental sample. Single-color control spectra are stored in the Spectral Reference Library. Unlike compensation controls, spectral references do not need to be acquired for every new experiment and can be reused. This allows users to create a personal or shared reagent library that simplifies experiment creation, saves time and valuable reagents, and increases efficiency. System Standardisation for Reproducible Data The ID7000 system can be operated in a Normal or Standardisation mode. Standardisation mode sets the system to an optimised master specification that allows researchers to maintain instrument settings between experiments and across multiple instruments to support longitudinal studies and cross-site collaboration. Standardisation mode adjusts the output of each channel so that the SSC and fluorescence detection sensitivities are the same between multiple instruments. PMT voltage correlation coefficients are calculated for each laser detection deck during daily QC, resulting in standardised measurements across all lasers, detectors, and instruments. Standardisation mode voltages can be adjusted synchronously or for each laser detection deck. The system supports standardisation of Area and Height data. This unique capability allows scientists to obtain the best data at maximum PMT voltages and reduces the need to rerun single-color controls for every experiment. Standardisation mode minimises subjectivity and instrument variability to yield highly reliable, accurate, and reproducible results. Designed for Automated Sample Acquisition Automation is present across the workflow to simplify operation and ensure accurate results. To give scientists more efficiency in their day-to-day work, the ID7000 comes with an advanced, best-in-class AutoSampler, which supports 5-mL tube racks (24 tubes), 96-well standard height, 96-well half deep, and 96-well deep plates, and 384-well standard flat bottom plates. Samples can be cooled to 4°C (39°F) using a Peltier element to prevent signal degradation and reduce variability. The sample in the individual tubes or wells can be agitated at the beginning of acquisition, intermittently or continuously, enabling stable, long-duration acquisition without sample settling. To ensure flexibility, the AutoSampler supports multiple wash options. A Dual Probe Wash option incorporates a cleansing mechanism that moves up and down to clean the inside and outside of the sample nozzle. Other wash options are available when speed of acquisition is the priority. The integration of intelligent software tools allows for management of bubbles and clogs, and integrated sensing mechanisms enable acquisition of critical samples at low volumes. The AutoSampler is equipped with an extra loading station for added convenience. Up to three 5-mL tubes can be placed in the extra positions and used, for example, for loading QC particles to streamline system QC. Or, the wash and cleaning solutions can be loaded and the software programmed for the system to shut down automatically after acquisition is completed. This is invaluable for users performing multi-sample acquisitions that are time consuming. Loading Options The ID7000 standard AutoSampler supports multiple loading options including a variety of 96-well and 384-well plates, as well as a 24-position tube rack for 12 x 75-mm (5-mL) tubes. AutoSampler Agitation Modes The ID7000 sample agitation feature is designed to keep large particles in suspension. The figure shows data with 30-µm particles when applying the initial mixing mode (Once) and the Cyclic mixing mode (Cyclic). It shows that the particles remain in suspension for 10 minutes when using the initial mixing mode, before there is an observed reduction in event rate. The Cyclic mixing mode, when used, continually mixes the sample throughout the entire acquisition, resulting in a consistent event rate with no settling. 5-mL Tube Rack Sample Cooling Over Time The graph above shows how sample temperature is maintained in the ID7000 AutoSampler when using a 24-position tube rack. Temperature measurement for tubes at multiple positions indicate that they maintain cooling over time when used in a 23°C ambient environment. AutoSampler Automated Workflows The ID7000 AutoSampler includes an extra tube station that accommodates up to three 12 x 75-mm (5-mL) tubes that can be used to support automated QC, system cleaning, and automatic shutdown workflows. Designed for True Ease of Use The ID7000 system software is easy to learn and use. It guides researchers from setup and QC to panel design, data acquisition, data analysis, through system shutdown. For accommodating users with varied experience levels, most workflows are designed with an expert mode and a guided mode. In guided mode, software wizards simplify system operation. The software provides access levels and tools to suit the needs of busy laboratories. Preferences allow users and administrators to specify options for overall instrument operation and experimental setup to facilitate use of the ID7000 spectral cell analyser. It is also possible to configure software preference settings for each individual user for added convenience. Intuitive Workflows for Acquisition and Analysis In the ID7000 system software, users can create experiments from a template, an existing experiment, a single-stained sample, or a multicolor assay to accommodate their preferred approaches. New users can create new experiments with the Experiment Design Wizard. To accommodate multi-sample and multiparameter experiment needs, the ID7000 software provides both a Shared Worksheet and an Individual Worksheet. A Shared Worksheet allows each sample in a group to use the same worksheet and thus makes experiment setup more intuitive. Instrument settings can be specified for each sample group and configured for lasers and detectors to give researchers the needed flexibility. Altering these settings for a sample in a group changes the settings for all unmeasured samples belonging to the same group. Instrument settings can be exported and imported
as needed.The acquisition functions are all accessible from the Acquisition tab and have been developed keeping a multi-laser workflow in mind. The tab accesses all functions that control the data acquisition, recording, and stopping, as appropriate, while monitoring the data acquisition status. To enable walkaway functionality, the system has an Event Checking function that can be set to stop or skip and continue automatic acquisition if the event rate drops or air bubbles are detected.
For sample analysis, the appropriate spectral references must be selected and fluorescence unmixing applied to remove the spillover components from each fluorescence channel. The unmixing can be manually adjusted on the screen using the Spectral Reference Adjuster. Making changes on one plot will result in the changes being applied automatically to all other plots having that fluorochrome as one of their parameters. Users can save the adjustment result as a new fluorescence unmixing matrix or can overwrite an existing fluorescence unmixing matrix.
The Analysis tab houses all controls for analysis operations on the ID7000 spectral flow cytometer, providing access to worksheet tools that let users choose how the data is displayed (such as plot types), and customize the display for their analysis needs. Plots and statistics provide real-time information during acquisition. The Experiment Explorer pane allows search by date and keyword, to display desired data on the worksheet. Once displayed, plots can be edited individually or as a group, to facilitate working with complex gating hierarchies.
The software also supports batch processes for users who need to run multiple samples, for operations like experiment setup, data acquisition and analysis, and export of data and statistics. Combined with the ability to program an automated shutdown, this enables scientists to focus on discovery instead of repetitive tasks.
Automated Setup and Quality Control to Ensure System Performance At startup, the Align Check and Performance QC wizards use beads to check instrument calibrations, to ensure the instrument is operating optimally. On-screen instructions guide the user through procedures, then display progress and report results. The QC results are available as a report that can be shared and archived. Trends over a desired period of time can also be displayed to ensure that the system performance is consistent as expected. At startup, the Align Check and Performance QC wizards use beads to check instrument calibrations, to ensure the instrument is operating optimally. On-screen instructions guide the user through procedures, then display progress and report results. The QC results are available as a report that can be shared and archived. Trends over a desired period of time can also be displayed to ensure that the system performance is consistent as expected. - Specifications
-
Optics / Performance
Excitation lasers 320 nm, 355 nm, 405 nm, 488 nm, 561 nm, 637 nm, 808 nm Detectors Photomultiplier tubes (32-channel PMTs, single-channel PMTs, and single-channel IR-sensitive PMTs) Fluorescence sensitivity (molecules of equivalent soluble fluorochrome)* FITC ≤ 58 MESF; PE ≤ 4 MESF; APC ≤ 7 MESF
*Specifications listed are an average of data obtained from multiple systemsPulse parameter Area, Height, Width (all channels) Signal resolution Height (20 bit), Area (32 bit)
Sampling frequency: 60 MHzDetectable particle size 160 nm with SSC Fluorescence resolution CV < 3% for the singlet peak of propidium iodide-stained Chicken Erythrocyte Nuclei (CEN) Event Rate 40,000 events per second Fluidics
Sample flow rate ~33 μL/min–200 μL/min Cleaning modes Priming, Probe Wash, Flow Cell Purge, Decontamination (with bleach), Hardware Shutdown (performed before system shutdown), Automatic Shutdown (programmed system shutdown after completion of Auto Acquisition) System fluidics 10-L capacity Sheath Tank and 10-L Capacity Waste Tank Sheath fluid (distilled water or phosphate buffered saline) System Software
Software ID7000 Acquisition and Analysis Software Supported file types exdat and FCS 3.1 AutoSampler
Sample loading options Multiwell plates: 96-well plate: standard height flat/V/U bottom, 96-well half deep V bottom, 96-well deep U bottom, 384-well standard flat bottom Tubes: 24-tube rack that accommodates 5-mL (12 x 75-mm) polystyrene/polypropylene tubes
Minimum sample volume* 5-mL tube: 100 µL in standard mode/50 µL in low-volume mode, 96-well plate: 55 µL in standard mode/10 µL in low-volume mode, 384-well plate: 40 µL in standard mode/10 µL in low-volume mode
*Minimum volumes provided for the sample vessel type are averages of data obtained from multiple experimentsCarryover < 0.1% (when measured with inner and outer Probe Wash) Throughput 96-well plate in 19 minutes (Based on an acquisition time of 2 seconds per well, no agitation, and no probe wash. The value reported is an average of multiple experiments.) Sample agitation Multiple mixing modes: Once at the beginning of acquisition, Cyclic, or Continuous Mixing Temperature Control From ambient temperature to 4°C with a Peltier sample cooling module Power consumption 600 W (max.) Workstation
Specifications Operating System: Windows® 10 Pro 64 bit
Processor: Intel® Xeon® 6248 2.5 GHz 20 Core
Memory: 256 GB DDR4
Graphics card: NVIDIA Quadro P2200
Storage: 8TB x3 (RAID5:16TB) HDD
2 Ethernet ports
4 display ports
DVD disc driveFacility requirements
Dimensions W: 41.7 in (106 cm) x D: 28.3 in (71.9 cm) x H: 29.9 in (76 cm) Weight 463 lb (210 kg) Power Consumption 900 W (2 independent 450-W circuits) Regulatory requirements
Class 1 Laser Product For non-clinical Research Use Only. Not for use in diagnostic or therapeutic procedures or for any other clinical purpose. United States: Class A digital device
Canada: CAN ICES-3 (A)/NMB-3(A)
EU: EU Declaration of Conformity 2006/42/EC (Machinery); 2014/30/EU (EMC)Laser configurations
Model No. of lasers, wavelength No. of detectors* LE-ID7000A 3LD: 405/488/637 FSC/SSC + 86F LE-ID7000B 4LD: 405/488/561/637 FSC/SSC + 112F LE-ID7000C 5LD: 355/405/488/561/637 FSC/SSC + 147F LE-ID7000D 6LD: 355/405/488/561/637/808 FSC/SSC + 149F LE-ID7000E 6LD: 320/355/405/488/561/637 FSC/SSC + 182F LE-ID7000F 7LD: 320/355/405/488/561/637/808 FSC/SSC + 184F *F = fluorescence
For Research Use Only.
Class 1 Laser Product. - Documents