SP6800 Spectral Analyzer
The Sony SP6800 Spectral Analyzer uses spectral technology to optimize sensitivity and enhance dim signal detection by collecting photons from 420nm to 800nm. Spectral technology also simplifies multicolor panel design, by eliminating bandpass filters and conventional compensation matrices while delivering better data and simplifying visualization for the study of heterogeneous populations.Novel global standardization mode automatically sets the system to a master specification with a single click. This capability eliminates instrument variability from day to day and across multiple instruments for greater reliability.
Advanced electronics and patented optical technologies bring simplicity to Spectral Analysis workflows. Sony’s patented Flowpoint™ core stability and tracking system and automated QC ensure the highest resolution possible of target populations.
The system features easy to use software that automates alignment and laser delay with set up wizards and simplified voltage settings. Each system includes FCS Express™ software in addition to Sony analysis packages to offer the highest flexibility in analysis.
The SP6800 is for non-clinical research use only and is a Class 1 laser product.
SP6800 System Analyzer
The SP6800 spectral flow analyzer improves sensitivity and simplifies application design, workflow, and analysis over conventional flow cytometers. This is achieved using spectral analysis technology, advanced electronics, and patented optical technologies.
These capabilities, unique to Sony Biotechnology systems, allow experienced and novice flow cytometrists to achieve greater flexibility for panel design and more accurate visualization of results.
The SP6800 software is easy to learn and use. It guides researchers from set-up to panel design, acquisition, analysis, and shutdown. User preferences allow users and administrators set up options for overall instrument operation and experiment set up to facilitate use and unattended operation procedures.
System Start Up
At start up Align Check and Performance QC wizards check instrument calibrations, using beads to ensure the instrument is operating optimally. On screen instructions guide the user through procedures, then display progress and report results. The performance report displays MESF, Q, and B values to describe real-time fluorescence detection performance. If desired, Align Check and Performance QC reports can be displayed in historical context.
Standardization mode sets the system to a master specification to eliminate variability in a single instrument or among instruments located across sites. This unique capability allows experiments performed on any Sony cell analyzer to produce highly reliable, accurate, and reproducible results. This function also eliminates setup subjectivity for collaborative or long term studies with different operators or experience levels across sites.
The Spectral Library lets users create a personal reagent library that simplifies experiment creation and saves time. An Acquisition Wizard assists the users with step by step instructions to acquire and analyze single positive controls for your spectral library. Once acquired the spectral reference for that reagent including the spectral index are available for future use. Information from the spectral library is available to users with a simple click improving accuracy and streamline panel design.
Experiments can be created using a template, an existing experiment, a single stained, or multicolor assay, in the Create Experiment window. Users can point and click to choose (or edit) an existing experiment and can easily select templates for wells or plates when creating a new experiment. A setup Assay Wizard guides users through the creation of a singlestained or multicolor assay simplifying experiment creation.
Acquisition Functions and Analysis
All acquisition functions, including instrument settings are controlled from the Acquisition Window. Worksheet tools let users choose how the data is displayed- (such as plot types), and customize for their analysis needs. Plots and statistics provide real-time information during acquisition. To increase sample acquisition to the cuvette, a variable booster lets users set acquisition speed from low (33ul/min) normal or high (250ul/min) offering flexibility.
FCS Express from De Novo Software
Spectral Analyzers from Sony include FCS Express, from De Novo Software. FCS Express offers a range of new analysis tools from live gating to batch analysis. Native support for Sony Spectral data files to enable sophisticated data transformations and visualizations such as spectral overlays, tSNE, Spade, and heat maps.
Dot plots and spectral plots can be set up with live gating. Live gating allows users to find positive and negative populations quickly and easily for added assurance about target populations.
Batch and Report
Spectral Analysis Technology
Spectral analysis technology is the foundation of the SP6800 system. Spectral flow cytometry streamlines workflow and yields better data by keeping all the light collected. In conventional systems, overlapping fluorescence is subtracted using color compensation, so less light is collected. Instead, spectral flow cytometers sum the fluorescence together and then use unmixing to mathematically separate the colors. This powerful capability also simplifies workflow including panel design, and improves visualization of autofluorescence. A unique prism collection system delivers emitted light to a 32 channel PMT. This produces 66 data points of signal detection for fluorescence and bright auto fluorescence to achieve accurate visualizations of fluorescent populations. This lets researchers see the complete spectral fingerprint of each fluorochrome from 420nm to 800nm.
A powerful capability of spectral technology is Unmixing. This allows researchers to separate fluorophores into pure signals that measure the quantity of each fluorophore at each pixel to more accurately measure data for analysis.
Spectral Unmixing separates each spectral fingerprint for complete and optimal visualization of fluorochromes.
Uniform measurement of Fluorescent Emissions
A correction system adjusts the offset and sensitivity of each channel of the 32 channel PMT to ensure a uniform and accurate measurement of fluorescent emission from 420nm to 800nm. The corrective system brings a standardization to all 32 PMTs ensuring the user is getting the most reliable data with the ease of adjusting only one voltage. This saves time over conventional flow cytometry operation where users must calibrate each individual PMT.
Subtracting Auto-Fluorescence Improves Visualization
In conventional flow cytometry cellular auto-fluorescence produced by pyridine (NAD/NADH), flavin (FMN, FAD), and other intracellular oxidative reactions can cause fluorescent signal contamination of other fluorescent markers. Other common sources of auto-fluorescence include cell fixation and permeabilization. Using spectral technology, auto-fluorescent spectral fingerprints can be subtracted to allow researchers to see the true fluorescent population.
Application data using 2 lasers
Fluorochromes: 405nm Laser
Sample data from Human PBMCs stained with seven markers conjugated to Brilliant Violet dyes offered by Sony Biotechnology. D. All populations were clearly resolved including fluorochomes with significant spectral overlap such as BV605 and BV650 (D).
*Except for LE-SP6800ZC (This HPCV value is calculated for red laser)