Flow Cytometry Equipment

Imaging and Flow Cytometry Core

Cell Sorter

BD FACSAria SORP

The BD FACSAria^TM SORP Cell Sorter is equipped with five lasers (Ultra-Violet 355nm, Violet 405nm, Blue 488nm, Yellow-Green 561nm and Red 640nm) which enables the detection of up to 18 fluorochromes. A gel coupled quartz cuvette flow cell in true fixed alignment with the laser, combined with trigon and octagon detection systems provide increased sensitivity and resolution.

A choice of nozzles (70, 86, 100 and 130 μm) allows optimization to a wide range of particle sizes. It is equipped with a versatile sort collection chamber that allows users to sort into up to 4-way 1.5- or 5-mL tubes, up to 2-way 15 mL tubes, as well as the ACDU unit which allows users to sort into 6-, 24-, 96-, 384-well plate or any custom collection device. Cooling and heating for sort collection are available.

Forward and side scatter properties allow for a minimum detectable particle size of 0.5 μm. The maximum data acquisition rate is 70,000 events/second and maximum sorting rate can go up to 7,000 events/second (at 28kHz). We recommend a minimum sample volume of 500 μL.

Applications:

Multi-color Cell Sorting (up to 18 colors)
Index Sorting
Single Cell Sorting

Detailed Configuration
FACSAria SORP – Basic Operation Procotol
Staff-in-charge: Ms. Wu, Si (Vincci) | 3917 9137 | flow.cpos@hku.hk

BD FACSAria Fusion

The BD FACSAria^TM Fusion Cell Sorter is equipped with four lasers (Violet 405nm, Blue 488nm, Yellow-Green 561nm and Red 640nm) which enables the detection of up to 16 fluorochromes. A gel coupled quartz cuvette flow cell in true fixed alignment with the laser, combined with trigon and octagon detection systems provide increased sensitivity and resolution.

A choice of nozzles (70, 86 and 100 μm) allows optimization to a wide range of particle sizes. It is equipped with a versatile sort collection chamber that allows users to sort into up to 4-way 1.5- or 5-mL tubes, up to 2-way 15 mL tubes, as well as the ACDU unit which allows users to sort into 6-, 24-, 96-, 384-well plate or any custom collection device. Cooling and heating for sort collection are available.

Forward and side scatter properties allow for a minimum detectable particle size of 0.5 μm. The maximum data acquisition rate is 70,000 events/second and maximum sorting rate can go up to 7,000 events/second (at 28kHz). We recommend a minimum sample volume of 500 μL.

Applications:

Multi-color Cell Sorting (up to 16 colors)
Index Sorting
Single Cell Sorting

Detailed Configuration
BD Aria Fusion – Basic Operation Procotol
Staff-in-charge: Ms. Wu, Si (Vincci) | 3917 9137 | flow.cpos@hku.hk

BD Influx with BSC

The BD Influx™ cell sorter is a 14-colour high speed cell sorter with four lasers (Violet 405nm, Blue 488nm, Yellow-Green 561nm and Red 640nm), an adjustable optical alignment and a pressure-driven fluidics system. It is a jet-in-air system and the nozzle assembly is designed to produce high droplet frequencies at relatively low pressures. This allows high-speed sorting while maintaining cell viability and functionality.

A choice of nozzles (70, 86, 100, 140 and 200 μm) allows optimization to a wide range of particle sizes. It is equipped with a versatile sort collection chamber that allows users to sort into up to 6-way 1.5-, 5-, 15- and 50-mL tubes as well as the CCDU unit which allows users to sort into 6-, 24-, 96-, 384-well plate or any custom collection device. Cooling and heating for sort collection are available.

Forward and side scatter properties (FSC and SSC) allow for a minimum detectable particle size of 0.2 μm. The maximum data acquisition rate is 200,000 events/second and maximum sorting rate can go up to 25,000 events/second (at 100kHz). We recommend a minimum sample volume of 300 μL.

Applications:

Rare Population Cell Sorting (up to 14 colors)
Index Sorting
Single Cell Sorting

Kindly be noted that ONLY Regular or Experienced user of FACSAria™ SORP/ FACSAria™ Fusion cell sorter who fulfills either one of the pre-requisites stated below is eligible for equipment usage due to the complicated operation of this cell sorting system:

a. Having large sample size which takes at least 4 hours to complete on FACSAria™ SORP/ FACSAria™ Fusion cell sorter.

b. Neither FACSAria™ SORP/ FACSAria™ Fusion can fulfill their experimental requirement.

Training of BD Influx™ Cell Sorting system will be held once per month based on first-come-first-serve principle. ONLY the training application of those who meet the prerequisite(s) stated above will be confirmed.
Detailed Configuration
BD Influx Standard Operation Protocol – Basic Operation
Staff-in-charge: Dr. Liu, Jing (Emily) | 3917 9136 | flow.cpos@hku.hk

BD Influx

The BD Influx™ cell sorter is a 16-colour high speed cell sorter with four lasers (Violet 405nm, Blue 488nm, Yellow-Green 561nm and Red 640nm), an adjustable optical alignment and a pressure-driven fluidics system. It is a jet-in-air system and the nozzle assembly is designed to produce high droplet frequencies at relatively low pressures. This allows high-speed sorting while maintaining cell viability and functionality.

A choice of nozzles (70, 86, 100 and 140μm) allows optimization to a wide range of particle sizes. It is equipped with a versatile sort collection chamber that allows users to sort into up to 6-way 1.5-, 5-, 15- and 50-mL tubes as well as the ACDU unit which zZallows users to sort into 6-, 24-, 96-, 384-well plate or any custom collection device. Cooling and heating for sort collection are available.

Forward and side scatter properties (FSC and SSC) allow for a minimum detectable particle size of 0.2 μm. The maximum data acquisition rate is 200,000 events/second and maximum sorting rate can go up to 25,000 events/second (at 100kHz). We recommend a minimum sample volume of 300 μL.

Applications:

Rare Population Cell Sorting (up to 16 colors)
Index Sorting
Single Cell Sorting

Kindly be noted that ONLY Regular or Experienced user of FACSAria™ SORP/ FACSAria™ Fusion cell sorter who fulfills either one of the pre-requisites stated below is eligible for equipment usage due to the complicated operation of this cell sorting system:

a. Having large sample size which takes at least 4 hours to complete on FACSAria™ SORP/ FACSAria™ Fusion cell sorter.

b. Neither FACSAria™ SORP/ FACSAria™ Fusion can fulfill their experimental requirement.

Training of BD Influx™ Cell Sorting system will be held once per month based on first-come-first-serve principle. ONLY the training application of those who meet the prerequisite(s) stated above will be confirmed.
Detailed Configuration
BD Influx Standard Operation Protocol – Basic Operation
Staff-in-charge: Dr. Liu, Jing (Emily) | 3917 9136 | flow.cpos@hku.hk

BD FACSDiscover S8

BD FACSDiscover S8 Cell Sorter

The BD FACSDiscover™ S8 Cell Sorter features BD CellView™ Image and BD SpectralFX™ Technology to expand the power of cell analysis and sorting by combining flow cytometry data with spatial and morphological insights.

This advanced cell sorter is equipped with five lasers (Ultra-Violet 349nm, Violet 405nm, Blue 488nm, Yellow-Green 561nm, and Red 637nm), six image detectors (3 blue laser scatter detectors and 3 fluorescence detectors), and 78 fluorescent detectors covering the spectrum from 365-860 nm.

Its fixed-alignment gel-coupled quartz cuvette flow cell eliminates the need for daily alignment and allows for quick nozzle changes. The sorter offers flexible sorting with different nozzle sizes (85, 100, and 130-μm) for optimizing a wide range of particle sizes. Additionally, it features a versatile sort collection chamber that enables sorting up to 6 ways into various tube sizes (1.5-mL, 2-mL or 5-mL) and plate formats (6-, 24-, 48-, 96-, 384-well), including microscope slides. Cooling and heating options for sort collection are available, and a Class II Type A2 biological safety cabinet (BSC) is included.

Forward and side scatter sensitivity enables separation of 0.16-µm polystyrene beads from noise. Image resolution resolves and provides image of 0.2-µm polystyrene particles from noise using at least one SSC parameter. The system can achieve a maximum data acquisition and sorting rate of 12,000 events/second using an 85-μm nozzle.

Applications:

Spectral Flow Cytometry Sorting or Analysis
Index Flow and Cell Imaging Data
Label-free Sorting
Fluorescent Localization
Cell Cycle Analysis, providing insight for Interphase, Prometaphase, Metaphase, Anaphase and Telophase.
Cell-cell Interaction
Functional Genomic Screening

Detailed Configuration

BD FACSDiscover™ S8 Cell Sorter – Basic Operation Protocol

Staff-in-charge: Dr. Liu, Jing (Emily) | 3917 9136 | flow.cpos@hku.hk

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Cell Analyzer

BD LSR Fortessa

The BD LSRFortessa™ is a 11-colour analyser with three lasers (Blue 488nm, Yellow-Green 561nm and Red 640nm), a fixed optical alignment and a pressure-driven fluidics system. The flow cell is a gel-coupled, and this combined with trigon and octagon detection systems provide increased sensitivity and resolution. This arrangement allows filter and mirror changes within the optical array to be made easily and requires no additional alignment for maximum signal strength, enabling researchers to identify cells with especially dim expression, or quantitate rare cell populations.

Forward and side scatter properties (FSC and SSC) allow for a minimum detectable particle size of 0.5 μm. The maximum data acquisition rate is 40,000 events/second. We recommend a minimum sample volume of 400 μL.

Applications:

Annexin V assay
Cell Cycle Analysis
Cell Proliferation assay
Multiplex bead assay for cytokines
Immunophenotyping

Detailed Configuration
LSR Fortessa – Basic Operation Protocol (Tube Mode)
Staff-in-charge: Dr. Liu, Jing (Emily) | 3917 9136 | flow.cpos@hku.hk

Agilent NovoCyte Quanteon

Agilent NovoCyte Quanteon analyzer is equipped with four lasers (Violet 405nm, Blue 488nm, Yellow-Green 561nm and Red 640nm) which enables the detection of up to 25 fluorochromes. The proprietary photomultiplier technology provides excellent sensitivity, stability, and a 7.2 log dynamic range, so that very dim signals and very bright signals can all be captured and gated in the same view. The NovoSampler Q is compatible with forty 5.0 mL 12 × 75-mm tubes or 24-/48-/96-/384-well plates and automatically loads them without intervention. Intuitive NovoExpress® Software is used in the NovoCyte® workstation.

Forward and side scatter properties allow for a minimum detectable particle size of 0.1 μm. The maximal event number for collection is 10⁷. We recommend a minimum sample volume of 50 μL.

Applications:

Annexin V assay
Cell Cycle Analysis
Cell proliferation assay
Multiplex bead assay for cytokines
Immunophenotyping
Small particle detection

Detailed optical configuration

ACEA NovoCyte Quanteon – Basic Operation Protocol
Staff-in-charge: Ms. Wu, Si (Vincci) | 3917 9137 | flow.cpos@hku.hk

Agilent NovoCyte Advanteon BVR

The NovoCyte Advanteon BVR analyzer is equipped with three lasers (Violet 405nm, Blue 488nm and Red 640nm) which enables the detection of up to 19 fluorochromes. The proprietary photomultiplier technology provides excellent sensitivity, stability, and a 7.2 log dynamic range, so that very dim signals and very bright signals can all be captured and gated in the same view. The NovoSampler Q is compatible with forty 5.0 mL 12 × 75-mm tubes or 24/48/96/384 well plates and automatically loads them without intervention. Intuitive NovoExpress® Software is used in the NovoCyte® workstation.

Forward and side scatter properties allow for a minimum detectable particle size of 0.1 μm. The maximal event number for collection is 10⁷. We recommend a minimum sample volume of 50 μL.

Applications:

Annexin V assay
Cell Cycle Analysis
Cell proliferation assay
Multiplex bead assay for cytokines
Immunophenotyping
Small particle detection

Detailed Configuration
Novocyte Advanteon BVR – Basic Operation Protocol
Staff-in-charge: Ms. Wu, Si (Vincci) | 3917 9137 | flow.cpos@hku.hk

Agilent NovoCyte Advanteon BVYG

The NovoCyte Advanteon BVYG analyzer is equipped with three lasers (Violet 405nm, Blue 488nm and Yellow-Green 561nm) which enables the detection of up to 15 fluorochromes. The proprietary photomultiplier technology provides excellent sensitivity, stability, and a 7.2 log dynamic range, so that very dim signals and very bright signals can all be captured and gated in the same view. It is compatible with 5.0 mL 12 × 75-mm tubes ONLY. Intuitive NovoExpress® Software is used in the NovoCyte® workstation.

Forward and side scatter properties allow for a minimum detectable particle size of 0.1 μm. The maximal event number for collection is 10⁷. We recommend a minimum sample volume of 50 μL.

Applications:

Annexin V assay
Cell Cycle Analysis
Cell proliferation assay
Multiplex bead assay for cytokines
Immunophenotyping
Small particle detection

Detailed Configuration
Novocyte Advanteon BVYG – Basic Operation Protocol
Staff-in-charge: Ms. Wu, Si (Vincci) | 3917 9137 | flow.cpos@hku.hk

BD FACSymphony A5 SE

The BD FACSymphony™ A5 SE is a 48 detectors analyser with five lasers (Ultra-violet 355nm, Violet 405nm, Blue 488nm, Yellow-Green 561nm and Red 640nm), a fixed optical alignment and a pressure-driven fluidics system. The flow cell is a gel-coupled, and this combined with Cascade array of square PMT detection systems provide increased sensitivity. This arrangement allows the system to gain full visible spectrum coverage to collect all light emitted without the need for filter changes, increasing dye flexibility and number of simultaneous colours, enabling researchers to study novel cell subpopulations which needs numbers of markers to identify.

Forward and side scatter properties (FSC and SSC) allow for a minimum detectable particle size of 0.5 μm. The maximum data acquisition rate is 40,000 events/second. We recommend a minimum sample volume of 400 μL.

Applications:

Multiplex bead assay for cytokines
High parameters immunophenotyping

Detailed Configuration
BD FACSymphony A5 SE – Basic Operation Protocol
Staff-in-charge: Dr. Liu, Jing (Emily) | 3917 9136 | flow.cpos@hku.hk

BD FACSymphony™A1

The BD FACSymphony™A1 features with four lasers (Violet 405nm, Blue 488nm, Yellow-Green 561nm, and Red 637nm) enabling detection of 16 colors. Each laser delivers 100 mW of power to excite samples in the flow cell made of rectangular quartz cuvette. Emitted light from the cuvette is delivered by fiber optics to the detector arrays. The optical pathways use signal reflection to maximize signal detection. In addition, it features the Small Particle Detector (SPD) ideal for small particle research.

It can achieve an acquisition rate of 8000 events/second. Meanwhile, Side Scatter (SSC) with the Small Particle Detector option, can separate 90-nm polystyrene beads from noise.

Applications: