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Moflo Astrios

Mon, 20 Dec 2010 10:53:47 -0500

Leveraging the historically stable fluidic design of the original MoFlo and the electronic processing of the MoFlo XDP, the MoFlo Astrios expands sorting capabilities with a full palette of laser options. Seven pinholes spatially separate each laser, providing flexibility and expandability. The Astrios also provides 6-way sorting, Touch Screen Control Panel, and comprehensive Biosafety features. The Astrios is also the first sorter with bead-less drop delay determination and monitoring.

iCyt Reflection

Sun, 02 Jan 2011 12:16:50 -0500

l Primary Fluorescence Collection Optic:

u Off-axis parabolic mirror (3x magnification)

u 95% reflection, 300 nm - 1 mm

u Directly coupled into two spatially separated fiber optics

l Secondary Fluorescence Optic (Optional):

u Custom objective coupled into three spatially separated fiber optics

u Transmission > 90%, 400 nm – 850 nm

l Optical Alignment: Automated

l Options:

u AOTF (Acousto-Optical Tunable Filter) with < 3 nm resolution

u 16 channel spectrometer (370 nm to 730 nm)

u Amplitude modulation (either directly or with EOM) from 5 to 40 MHz

u Secondary fluorescence collection optic

u Sort deposition system

u Biohazard containment: Baker SterilGard . III Advance hood integration

l Dimensions:

l Research Reflection Instrument (W x D x H)

u 45 x 33 x 57 (inches)

u 114 x 84 x 145 (cm)

l DPA (Digital Processing Assembly) (W x D x H)

u 30 x 36 x 32 (inches)

u 76 x 91 x 81 (cm)

l FDA (Fluorescence Detection Assembly) (W x D x H)

u 27. x 16. x 13. (inches)

u 70 x 42 x 34 (cm)

l Electrical requirements: (2) 20 A 120V dedicated lines

l Pressure (5 – 100 psi): House air, nitrogen or optional Jun-Air compressor

l Vacuum (12 inches of Hg): House or optional Jun-Air compressorl HAPS (Highly Automated Parallel Sort) modules: 1 to 4 per instrument

l Analysis Rates: greater than 150,000 events per second for each HAPS

l Sort Rates: greater than 70,000 events per second for each HAPS

l Sort Purity: >99%

l Drop Drive Frequency: Up to 120 kHz

l Sort Directions:

u 32 programmable sort streams

u Separate streams for empty drops and non-sorted cells

u Up to 4 streams collected into tubes

u Tubes supported 0.6 ml, 1.5 ml, 5 ml, 15 ml, 50 ml

u Up to 32 discrete streams for either indexed or proportional sorting

l Fluorescent Detectors: Up to 62 PMTs

l Wide Angle Forward and Back Light Scatter: Up to 8 PMTs

l Narrow Angle Forward Light Scatter: Up to 4 photodiodes

l Streaming ADC inputs (105 MHz Sampling Rate, 14 Bit Resolution): Up to 64 channels

l Listmode Parameters:

u Up to 256

u Pulse features (area, height, width, rise time)

u Sort decisions (cell sort classification, droplet stream classification)

u Time interval between pulses

u Custom programmable pulse features

l Fluorescence Compensation: Full matrix among all features

l Simultaneous Laser Excitation Lines: Up to 7 (355, 405, 488, 532, 561, 592, 640 nm)

l Laser Options: Any vendor, any configuration with industry standard optical table

l Nozzle Sizes: 50 μm to 200 μm

l Measurement Dynamic Range: Greater than 10,000 x

l Measurement Precision: CV of < 1%

l Narrow Forward Scatter Collection Angle: 1 to 5 degrees half-angle

l Optional Wide Forward Scatter Collection Angle: 5 to 41.3 degrees half-angle

l Number of Analysis Regions: Greater than 512

l Number of Separate Sort Gates (Based on Boolean Expressions of Region): Up to 256

Becton Dickinson FACSAria I, II, and III

Thu, 16 Dec 2010 14:37:43 -0500

Excitation Optics Optical Platform Fixed optical alignment of all Class IIIb lasers upon the cuvette flow cell. The 488-nm and 633-nm lasers come stan- dard. All other laser choices are optional. All are solid state except the 633 nm, which is gas.

Beam height: 9 ±3 μm Beam width: 65 ±7 μm

Power Out of the Laser Head

375 nm: >7 mW, top hat shape 405 nm: >50 mW, elliptical shape 445 nm: >20 mW, elliptical shape 488 nm: >20 mW, elliptical shape 561 nm: >50 mW, elliptical shape 633 nm: >18 mW, elliptical shape

Emission Optics Optical Coupling The quartz cuvette flow cell is gel-coupled by refractive index-matching optical gel to the fluorescent objective lens for optimal collection efficiency.

Numerical aperture: 1.2

Forward Scatter Detector and Filters

Photodiode with 488/10 bandpass filter for the 488-nm laser and 462/52 band- pass filter for the 445-nm laser.

Side Scatter Detector

Photomultipler with a 488/10 bandpass filter for the 488-nm laser and a 445/15 bandpass filter for the 445-nm laser.

Fluorescence Detectors and Filters

Five fixed-fiber apertures for the 488, 633, 561, 445, and 405-nm lasers

The 445-nm laser focuses on the flow cell at the same location as the 488-nm laser. The 375-nm laser is air launched and focuses on the flow cell at the same location as the 405-nm laser. An octagon technology detector array enables user- defined detection configurations.

Wavelengths detected from 488-nm laser without 561-nm laser installed • 530/30-nm FITC • 585/42-nm PE

• 616/23-nm PE-Texas Red® • 695/40-nm PerCP-CyTM5.5 or PI or

675/20-nm PerCP-Cy7 • 760/60-nm PE-CyTM7

Wavelengths detected from 488-nm laser when 561-nm laser installed • 530/30-nm FITC • 695/40-nm PerCP-Cy5.5 or PI or

675/20-nm PerCP

Wavelengths detected from 633-nm laser

• 660/20-nm APC • 780/60-nm APC-Cy7 or APC-H7

Wavelengths detected from 405-nm laser

• 450/50-nm BD HorizonTM V450, Pacific BlueTM, DAPI

• 510/50-nm BD Horizon V500, AmCyan

Wavelengths detected from 561-nm laser

• 582/15-nm PE • 610/20-nm PE-Texas Red®, Living

Colors®, mCherry, propidium iodide • 670/14-nm PE-CyTM5 or 710/50-nm

PE-Cy5.5 • 780/60-nm PE-Cy7 Wavelength detected from 445-nm laser • 510/80-nm, cyan fluorescent protein

(CFP)

Wavelengths detected from 375-nm laser

• 450/20-nm Hoechst Blue, DAPI • 670 LP Hoechst Red, PI

Filters and mirrors are user changeable. Additional detectors up to a total of 18 wavelengths can be added to the arrays.

Steering Optics

488, 633, 561, 445, and 405-nm laser: Fiber optics steer the fixed alignment laser beams onto the expansion prisms to focus them on the cuvette flow cell.

Near UV (375-nm) laser: Air launched and focused on the cuvette flow cell

Sort Performance Drop Drive Frequency Range 1–100,000 Hz

Purity and Yield

At 70 psi and 90 kHz with an average threshold rate of 25,000 events per second, a four-way sort achieved a purity of >98% and a yield >80% of Poisson’s expected yield. Higher threshold rates

up to 70,000 events per second can be achieved without affecting purity; however, yield will decrease based on Poisson statistics.

Viability

Sorts were performed at a variety of sheath pressures using cell lines and human peripheral blood mononuclear cells (PBMCs). All sorts resulted in cells that proved viable and proliferated for several days post-sort.

Sort Collection Devices

Two-way sorting: 12 x 75 mm, and 15 mL

Four-way sorting: 1.5 mL microtube and 12 x 75 mm

Sort Collection Cooling

Water recirculator for refrigeration/ heating (optional)

BD FACSTM Accudrop

• Red diode laser provided for fully automated drop-delay determination

• Automated drop breakoff monitoring

• Automated clog detection and sort tube protection system using Sweet Spot technology

Loading Sample Input Sizes Microtubes, 12 x 75 mm, and 15 mL

Polystyrene or polypropylene tubes can be used.

Sample Input Agitation

Adjustable through the software to keep sample constantly suspended

Temperature Control

Sample input, software-adjustable: 4, 20, 37, and 42°C

Sample output for sort collection: water recirculation unit (optional)

iCyt Synergy

Fri, 17 Dec 2010 18:26:35 -0500

Configuration

The Synergy system can be configured as a single channel

cell sorter or analyzer, dual channel sorter or analyzer, or a

sorter/analyzer combination.

Standard bench-top model dimensions:

48" w x 26" d x 30" h

Bio-Safety (BSC) model dimensions:

53" w x 48" d x 89" h

Operation

Sorting modules can be configured for either jet-in-air or

gel-coupled sense-in-channel operation.

Sorting nozzle sizes include 70 μm, 85 μm, 100 μm, 130 μm

Accommodates particles up to 40μm

Provides 4-way sorting into most popular collection tube

formats. Optional Sort Deposition System (SDS) supports

sorting into most popular plate formats.

Analyzer modules are configured for closed-loop gelcoupled

sense-in-channel operation. Automated sample

delivery system supports most popular plate and tube

formats.

Excitation

Each sorter or analyzer module can be configured with up

to eight lasers focused on five spatially separated

interrogation points.

Standard laser options

Standard laser options include Omicron LuxX™ 375nm,

405nm, 445nm, 488nm, 642nm diode lasers and Cobalt

Calypso™ 491nm, Samba™ 532nm, Jive™ 561nm,

Mambo™ 594nm DPSS lasers.

Standard beam size is approximately 15 μm x 60 μm.

Beam spots are spatially separated by more than 350 μm

to eliminate laser crosstalk.

Collection

A multi-interrogation point Reflective Collection Optic

(RCO) provides exceptional fluorescence sensitivity,

resolution, and spatial isolation with an effective NA of 1.2

at five beam spots. Fiber optic cables deliver light to userconfigurable

PMT arrays.

Each sorter or analyzer module can be configured with up

to 32 detectors: up to 26 fluorescence PMTs and 6 scatter

diodes.

PMT arrays can be configured to detect all popular

fluorochromes and fluorescent proteins including:

375nm - DAPI, Hoechst Blue/Red

405/445nm - CFP, BFP, Pacific Bue, Pacific Orange, QDots,

488nm - GFP, YFP, FITC, PE, PE-TR, PI, PerCP-Cy5.5, PE-Cy5.5,

PE-Cy7

532/561nm - RFP, dsRed, mCherry, PE, PE-TR, PI, PE-Cy5,

PE-Cy5.5, PE-Cy7

594/642nm - Alexa Fluor® 594, APC, Alexa Fluor® 700,

APC-Alexa Fluor® 750

Processing

Each SynergyTM sorter and analyzer module can acquire

up to 24 simultaneous parameters with 105 MHz sampling

at up to 100,000 events per second.

Linear area, linear height, linear width, log area, log

height, and log width pulse features are available.

Full color- compensation matrix via WinList acquisition and

analysis software.

Sense-in-channel sorting at up to 35,000 events per

second.

Jet-in-air sorting at up to 70,000 events per second.

Performance

Forward Scatter resolves 0.5μm diameter particles from

background.

Fluorescent Sensitivity - Measured using Sphero™ Rainbow

Particles RCP-30-5A:

FITC < 125 MESF

PE < 100 MESF

Sort Performance

Purity > 99%

Yield > 90% of theoretical based on Poisson statistics

Becton Dickinson Influx

Fri, 17 Dec 2010 14:15:59 -0500

Innovative Fluidics Design

The BD Influx™ fluidics system features a unique acoustical coupling in the nozzle assembly to reliably create droplets for sorting, while ensuring low shear stress to optimize cell viability, even at high pressures. To support aseptic sorting, the fluidics path is easily removed and replaced with the optional disposable fluidics kit.

Intuitive Alignment

The optical system that directs lasers to the interrogation point is easy to use, intuitive, and adaptable. Up to seven laser paths can be configured. Wavelength options include 355-nm, 405-nm, 457-nm, 488-nm, 532-nm, 561-nm, and 640-nm lasers.

A range of standard detection modules is available to configure the system based on application needs.

Flexible Sorting

In addition to two-, four-, and six-way sorting, the BD Influx™ system features special sort modes such as plate, proportional, and positional sorting, which can be optimized for application requirements.

Environmental Control

The BD Influx™ aerosol management system draws air away from the operator through a HEPA filter, reducing the amount of aerosols in the sort chamber.

BD Influx Software: From Acquisition to Sorting and Analysis

BD Influx™ software provides comprehensive control of the cell sorter from configuration and compensation setup to acquisition, sorting, and analysis, making it easy to customize the instrument for specific applications.