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SH800S
Cell Sorter

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Sorting Made Simple™

The benchtop SH800S cell sorter permits sorting of a wide range of cell sizes for many applications using the 70-μm, 100-μm, and 130-μm microfluidic sorting chips. This novel, chip-based design is fully integrated with comprehensive fluidics controls and advanced automation for set-up, acquisition, sort and analysis to make sorting less subjective, more precise and easier to perform.

System software is intuitive and supports sorting into tubes and 96- and 384-well plates. The software generates FCS 3.0 and FCS 3.1 files that can be exported to third party analysis tools. The SH800S is for non-clinical research use only and is a Class 1 laser product.

The SH800S is flexible and can support a variety of applications for individual and core labs. For applications in which cross-contamination is a concern, optional e-beam sterilized consumables can be used to replace the sample line and sorting chip. An optional class A2 Level II biosafety cabinet provides protection for personnel and products. The biosafety cabinet was custom designed for the SH800S and tested by The Baker Company to meet international standards. The optical design offers up to four collinear excitation lasers (488 nm, 405nm, 561nm and 638nm) and six fluorescence detectors. The six free-form PMTs enable detection of fluorescence signals from any laser based on filter selection.

Automation from Set-up to Analysis

The SH800S provides the highest level of automation and ease of use available in a cell sorter. Researchers can accurately set up, calibrate, and monitor sorting with a push of a button. This delivers true ease of operation to save time and improve consistency of results.

A novel microfluidic sorting chip uses patented CoreFinder™ technology to automate key steps of instrument setup and operation, streamlining workflow.

The system is versatile, allowing researchers to sort of a wide range of cell sizes to accommodate application requirements. Chip sizes include 70 μm, 100 μm, and 130 μm.

sh800-system-automated-chip-loading

Automated Chip Loading and Positioning

System set-up begins with a one-touch install to load the microfluidics flow cell chip. Actuators ensure precise positioning of the chip inside the chip loader. Sheath, sample, and vacuum lines connect and seal automatically to their respective ports once the chip is loaded. More at sonybiotechnology.com
ma900_automation_axis_adj

Automated Optical Axis Adjustment

The alignment of the chip to the lasers is optimized automatically during setup using the Sony patented Blu-ray™ technology for aligning and tracking laser position. On a daily basis, using AutoSetup beads, the X and Z positions of the chip are adjusted to ensure consistent results.
sh800-system-automated-side-stream

Automated Side Stream Calibration

The angle and the position of the side streams are calculated and adjusted during setup for tube and plate sorting. This ensures that the sort stream is centered in the collection tube automatically without manual adjustment.
sh800-system-automated-drop-delay

Automated Drop Delay Calibration

A dedicated laser and camera perform real-time analysis of droplet images using AutoSetup beads. Patented technology calculates drop delay by capturing the fluorescence image of the drop-delay beads. More at sonybiotechnology.com

Automated Sort Monitoring and Droplet Calibration

The SH800S software monitors and actively makes adjustments to the drop drive to maintain a stable breakoff point. This feature ensures consistently good sort performance and allows detection of clogs and empty tubes, and facilitates walk-away operation.

The droplets are automatically calibrated by adjusting the frequency and the drop drive to achieve optimal break off point (BOP) for each type of sorting chip.

Software

Software, sensors, and CoreFinder patented technology provide automation across the workflow from set-up to shutdown to ensure consistency, save time and improve the accuracy of results. The interface is designed with ribbon tabs that logically organize features to make them rapidly accessible. An experiment-centric approach makes the software easy to teach, learn, and use.

Software wizards include step-wise workflows that guide users through start up, multicolor compensation, sorting, and system cleaning. Since the system simplifies even advanced sorting techniques, it can be used by researchers with little sorting experience.

System Startup

Upon start up, the system initialization includes diagnostics that ensure all subsystems are properly working. Once verified, the system status and a green ready message are displayed on the LCD monitor on the front of the instrument.

sh800-software-startup
The LCD monitor on the front of the SH800 displays status information during operation.

Initial Instrument Set-up, Precision Alignment, and Calibrations

The set up wizard guides researchers through the process of loading a sorting chip, selecting lasers, and inserting the optical filters required.

Fluidics Check

When complete, the fluidics check starts, and sheath fluid droplets appear from the tip of the sample probe. If needed, de-bubble and sample line cleaning wizards can be run. Once complete, the auto calibration screen is displayed.

Alignment and Calibrations

The system automatically and precisely aligns the sorting chip to the lasers, calibrates the droplets, and side streams and estimates the drop delay using AutoSetup beads.

Automatic calibration performs precise measurements using the side stream monitor. The software displays progress and status at each stage of calibration.
The QR code on the packaging is scanned to identify the chip information, including nozzle size.
The sorting chip is easy to load.
To further simplify set-up, the Fluorochrome Detection Matrix screen displays where to insert optical filters.

Experiment Settings

Users can create an experiment by selecting a new template, a recent experiment, or a shared template (public) from the Experiment window. If a new template is selected, dialogs guide the user to choose experiment settings such as sample groups, tubes, and pulse parameters for data acquisition. Once a template is selected, the user can start the acquisition or compensation wizard.

Selecting a template or a recent experiment on the left of the window displays the experiment structure on the right.

Data Analysis and Display

Data is displayed as dot plots and histograms on worksheets, and events can be marked using gates. The software has a number of tools to select, adjust, label, and measure statistics of target populations. Data can be easily exported as FCS formats (3.0 or 3.1 ) to use with third party analysis software.

The Experiments tab provides easy access to all lists of previous experiments as well as visualization and statistics tools for analysis.

Sorting

Once events are gated on plots, targeted cells can be sorted for further analysis. Sort control is managed in a simple dialog where settings and configurations are selected. The SH800S supports a variety of sorting methods and vessels including 2-way tubes, 8-well strips, slides, and 6-, 12-, 24-, 48-, 96- and 384-well plates.

The system supports eight sorting modes to provide different levels of purity (including single-cell mode) and yield to enable optimal results. In addition, the software provides an assisted mode for large cells to improve sorting performance. Three sorting chip sizes (70 μm, 100 μm and 130 μm) are available to accommodate a range of cell sizes and applications.

Biosafety Cabinet

The BCC300AMS Class II biosafety cabinet custom designed for SH800S by the Baker Company is available as an option to provide personnel and product protection. The cabinet measures 118 cm (W) x 99.1 cm (D) x 224 cm (H).

Biosafety Safety Standards Compliance

The cabinet was tested by The Baker Company using microbiological assays with the SH800S sorter inside the work area. The testing concluded that the biosafety cabinet with the SH800S inside met several international biosafety standards including the National Sanitation Foundation Standard 49 (NSF49), and the European Standard 12469.

Built-in Aerosol Management

The cabinet incorporates a built-in aerosol management system which operates independently to actively evacuate aerosols from the sort collection chamber. The dual routes of aerosol evacuation ensure maximum personnel protection.

More at sonybiotechnology.com

SH800S Cell Sorter applications

Resolution and Sensitivity
A. Fluorescence sensitivity MESF values measured using Spherotech® 8 peak-beads are: FITC 120 and PE 110. B. Fluorescence resolution was measured using propidium iodide (PI) stained chicken erythrocyte nuclei (CEN). A coefficient of variation of <2.5% was observed for the G0/G1 peak.
Immunophenotyping Assays
Distinct resolution of multicolor samples is seen with a four laser SH800S system. A. Normal human blood was stained with CD3 Alexa Fluor® 488, CD4-PECy7, CD8 Alexa Fluor® 700, CD19 APC, CD16+CD56, PE and HLA-DR BV421. All plots were gated on lymphocytes. B. High-purity sorting of the regulatory T cells (CD3+ CD4+ CD25 high CD127-) population. iImmunophenotyping assays

Gates and Statistics

Immunophenotyping Assays
NameEvents%Parent%Total

All Events

30,0000.00%100%

Lymph

18,06760.22%60.22%

CD3+4+

9,18150.82% 30.60%

Lymph

602 6.56% 2.01%

Fluorescent Protein Analysis
Human cell lines co-expressing GFP, YFP, dsRed and CFP using the fluorescent protein optical filters are shown. Refer to the filter guides for the fluorescent protein filter sets.
fluorescent protein analysis
Sorting and Viability Analysis
These plots show a sorting and viability analysis of differentiating mouse embryonic stem cells expressing Isl1Cre-tdTomato. Analysis of the tdTomato (+) purified population is shown. the inset image (E) shows cells in culture 24 hours post sorting. sorting and viability analysis
Small Particle Resolution
High-speed sorting of E.coli using a 70-µm sorting sample of cells was sorted at a threshold event rate of 30,000 events/sec at a sort setting of 50 kHz and 40 psi. Post-sort analysis shows that >95% purity was obtained. small particle resolution

SH800S Cell Sorter specifications

SH800S cell sorter instrument specification
Optics
Excitation lasers488 nm, 405 nm, 638 nm and 561 nm laser
Output power30 mW (max.) optical fiber output
Beam alignmentCollinear optical system
Detection parameters6 fluorescence + 2 scatter
Analog-to-digital converters (ADC)8-channel 20-bit, 110MHz
Pulse measurementHeight, Area, Width
Fluidics
Sample tubeSingle, auto-loading tube
Tube types0.5-mL, 1.5-mL, 5-mL, and 15-mL tubes
Sort devices2-way tube, multiwell plates, PCR tubes, slides
Temperature control5°C, 37°C (electric cooling method)
Agitation unitEccentric rotation
Magnetic drive300-rpm speed
Sorting chip size70 μm, 100 μm and 130 μm
Sort Performance
Event rate100,000 eps
Sorting speedUsing the 70-μm sorting chip at 50 kHz, an average threshold of 12,000 events per second can be achieved with >98% purity and >80% yield. A threshold rate up to 30,000 events per second can be achieved without affecting purity but with a decrease in yield based on Poisson’s statistics.
Scatter resolution0.5 μm
Fluorescence resolution< 2.5% half-peak coefficient of variation (HPCV)
Fluorescence sensitivityFITC 120 MESF, PE 110 MESF
Ancillary
DimensionsW: 21.7 in. (55 cm)x D: 21.7 in. (55 cm)x H: 28.4 in. (72 cm)
Fluidics cartW: 30 in. (78.6 cm) x 20.5 in. (52.1 cm) x 22.8 in. (58.0 cm)
Weight216 lb (98 kg)
Fluidics cart71 lb (32 kg) (dry weight)
LCD panel7-inch, 800 x 480 pixels
Power supply100-240 V, 50/60 Hz
Power consumption500 W (max.)
Operating temperature17.5°C to 27.5°C
Relative humidity20% to 80%
Compliance:
Environmental
Rohs
Operating systemMicrosoft® Windows® 8 Professional, 64 bit
Data file structureFlow Cytometry Standard (FCS) 3.0 or 3.1
Safety standards complianceUL, CE, CSA

Filter Guide

Fluorochrome Guide

sh800s-cell-sorter-specifications-filter-set1sh800s-cell-sorter-specifications-filter-set2

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