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Cytek Aurora

Overview
Technology
Specifications
Software
Plate Loader
Reagents
Performance Data
Why Aurora?
Publications

Overview


A prodigy incorporating a unique combination of patent-pending innovative technologies that takes flow cytometry to the next level of performance and flexibility.

With up to four lasers, three scattering channels, and 48 fluorescence channels, the Aurora suits every laboratory’s needs, from simple to highcomplexity applications. A paradigm shifting optical design provides unprecedented flexibility, enabling the use of a wide array of new fluorochrome combinations without reconfiguring your system for each application. The state-of-the-art optics and low-noise electronics provide excellent sensitivity and resolution. Flat-top beam profiles, combined with a uniquely designed fluidics system, translate to outstanding performance at high sample flow rates.

The end result is a system that delivers high quality data where rare and dim populations are easily resolved, regardless of assay complexity.

SpectroFlo® Software offers an intuitive workflow from QC to data analysis with technology-enabling tools that simplify running applications.

The Cytek team has reimagined every aspect of cytometry hardware and software to deliver an instrument that fulfills every scientist's needs.


Maximum Channels

51 channels of detection over the full emission spectra.


Maximum Colors

Up to 24 colors demonstrated including fluorochrome with emission spectra in close proximity to each other.


Maximum Sensitivity

Sensitivity redefined using state-of-the-art optics and low-noise electronics.


Maximum Flexibility

No changing optical filters for any fluorochrome.

Use any commercially available fluorochrome excited by the onboard lasers.


Maximum Accessibility

A powerful, high value system that is accessible to a wide range of users.

Technology


Aurora's Revolutionary Technologies:

From Vision to Reality

The Aurora is capable of up to 51 detection channels (48 fluorescent channels, FSC, blue laser SSC, and violet laser SSC) and is empowered by revolutionary technologies, including:

  • Proprietary high sensitivity Coarse Wavelength Division Multiplexing (CWDM) semiconductor detector arrays, enabling more efficient spectrum capture for dyes emitting in the 420-830 nm range.
  • High bandwidth electronics design scalable beyond 51 channels.
  • Robust vacuum fluidics system enables ultimate flexibility in sample input formats.
  • Exceptional small particle detection is enabled by violet laser scatter, narrow beam height, and proprietary flat top laser design.

Specifications

Optics

Excitation Optics

Optical Platform

Aurora contains a fixed optical assembly with the capacity to be configured with up to five spatially separated laser beams. Laser delays are automatically adjusted during instrument QC.

Lasers

Three laser configuration: 405nm: 100mW, 488nm: 50mW, 640nm: 80mW
Four laser configuration: 405nm: 100mW, 488nm: 50mW, 561nm: 50mW, 640nm: 80mW

Beam geometry

Flat-Top laser beam profile with narrow vertical beam height optimized for small particle detection.

Emission Optics

Emission Collection

Fused silica cuvette coupled to high NA lens for optimum collection efficiency to optical fibers.

Forward and Side Scatter Detection

FSC: high-performance semiconductor detector with 488nm bandpass filter.

Violet SSC: high-performance semiconductor detector with 405nm bandpass filter.

Fluorescence Detectors

Proprietary high sensitivity Coarse Wavelength Division Multiplexing (CWDM) 16-channel semiconductor detector array per laser enabling more efficient spectrum capture for dyes emitting in the 400-900 nm range. No filter changes required for any fluorochrome excited by the 405nm, 488nm, and 640nm lasers.

Standard Optical Configuration

Violet detector module: 16 channels uneven spaced bandwidth from 420nm-830nm.

Blue detector module: 14 channels uneven spaced bandwidth from 500-830nm.

Red detector module: 8 channels uneven spaced bandwidth from 650-830nm.

Yellow-Green detector module (in four laser systems only): 10 channels uneven spaced bandwidth from 570-830nm.

Fluidics

Sample Flow Rates

Low: 15 µL/min, Medium: 30µL/min, High: 60µL/min

Fluidic Modes

Long clean, SIT flush, Purge filter, Degas flow cell, Clean flow cell

Manual Sample Input Formats

12x75mm polystyrene and polypropylene tubes

Standard Fluidic Reservoirs

4L fluid container set with level-sensing provided. Compatible with 20L sheath and waste cubitainers.

Plate Loader Option

96-well microtiter plate capability

Sample Dead Volume

5μL for 12x75mm tube

Performance

Fluorescence Sensitivity

FITC: <110 MEFL, PE: <35 MEFL, APC: <15 MEFL, Pacific Blue: <200MEFL

* measurements based on an average from three systems and performed using SPHERO Rainbow Calibration Particle (RCP-30-5A) based on its peak emission channel.

Fluorescence Linearity

FITC R2 ≥0.995 / PE R2 ≥0.995

Forward and Side Scatter Resolution

Performance is optimized for resolving lymphocytes, monocytes, and granulocytes.

Side Scatter Resolution

Capable of resolving 0.1µm beads from noise.

Carryover

<0.1%

Data Acquisition Rate

35,000 events/s

Software

SpectroFlo® Software

Live unmixing during acquisition

Developed specifically to streamline assay setup, data acquisition, and file export.

Automated QC module

Autofluorescence extraction

Raw and Unmixed FCS 3.1 files

Electronics

Signal Processing

Digital signal processing with automatic window gate adjustment.

22-bit 6.5 log decades.

Threshold using any single parameter or combination of parameters.

Pulse Shape Parameters

Pulse Area and Height for every parameter. Width for scatter parameters and one fluorescence parameter for each laser.

Workstation

Operating System

Windows® 10 Pro 64-bit

Processor

Intel® Core i7 processor, 3.6 GHz

RAM

32GB

Hard Drive

500GB SSD / 1TB SATA

Video Processor

NVIDIA® GeForce

Monitor

32” UHD 4K Monitor

Installation Requirements

Dimensions (W x D x H)

Instrument Dimensions

Without loader: 54 x 52 x 52 cm
With loader: 58.4 x 62 x 52 cm

Instrument Weight

Instrument weight (4 lasers): 80 kg
Loader weight: 13kg

Computer Dimensions

29.1 x 9.25 x 34.4 cm

Recommended Workspace

157 x 71 x 132 cm

Room Requirements

Power

100-240V, 50/60 Hz, 2A max

Heat Dissipation

500W with all solid-state lasers

Temperature

17–28°C

Humidity

20%-85% relative non-condensing

Air filtering

No excessive dust or smoke

Lighting

No special requirements

Regulatory Status

For Research Use Only. Not for use in diagnostic or therapeutic procedures.

Documents


Cytek Aurora Product Brochure

Click below to download the complete product brochure for the Aurora. It contains everything you need to know about this revolutionary product.

Download PDF

Software


SpectroFlo® Software Guided Workflows

The new SpectroFlo® software offers an intuitive workflow from QC to data analysis with technology-enabling tools that simplify running any application.

QC and Setup:

Run Daily QC to monitor instrument performance and add reference controls.

Library:

Add or remove experiment templates, worksheet templates, fluorochrome information, QC bead information, and more.

Getting started with the new SpectroFlo software.
Extra Tools:

Unmix data using controls from different experiments or apply virtual filters to your data.

Users:

For administrative controls.

Preferences:

Customize the software appearance. Set default plot sizes, text sizes and fonts, gate colors, print layout, statistics table options, and more.

Acquisition:
SpectroFlo software Experiment Menu

Experiment Menu

SpectroFlo software Worksheet Menu

Worksheet Menu

SpectroFlo software Fluidics Menu

Fluidics Menu

SpectroFlo software Plate Calibration

Plate Calibration


Experiment Workflow:

From the Acquisition menu, you can start a new experiment and get to your data in three simple guided steps.

Step 1: Create Your Experiment

Create your experiment, choose fluorochromes, and add labels, tubes, worksheets, and stopping criteria in this guided workflow.

Step 2: Acquire Your Tubes

Load and acquire your samples.

Step 3: Unmix Your Data

Visualize your reference controls spectra using our unmixing wizard.

Plate Loader


Get to Know Our Automatic Micro-Sampling System (AMS)

Cytek's Automatic Micro-Sampling System (AMS) with the Aurora

Meet the New AMS

The new AMS offers preset and user adjustable settings that allows the loader to be fine tuned to your experimental requirements. The AMS is specifically designed to streamline experimental workflow and seamlessly integrates into the Aurora. The AMS also offers ease of use, low carry over, and minimal dead volume.

Quick and easy

Reliable 96 well plate acquisition maximizes productivity.

Easily change between plates and tubes in a matter of seconds.

Three throughput modes

Optimized acquisition speeds, from low carry over to high throughput.

User customizable modes

Fully customizable with a suite of user modes to fit a variety of applications and workflows.

Performance Data


More Choice, Greater Flexibility, Easier Setup

The optical design combined with the unmixing capability in SpectroFlo® software allows greater fluorochrome choice, panel flexibility, and easy setup without having to change filters. The three laser configuration provides outstanding multi-parametric data for a wide array of applications. Markers and fluorochromes in a 24-color panel designed for identification of circulating cell subsets in human peripheral blood are summarized in the table below:

Specificity Fluorochrome Specificity Fluorochrome Specificity Fluorochrome
CCR7 Brilliant Violet 421 CD11c BD Horizon BB515 CD27 APC
CD19 Super Bright 436 CD45RA Alexa Fluor® 488 CD123 Alexa Fluor® 647
CD16 eFluor® 450 CD3 Alexa Fluor® 532 CD127 BD Horizon APC R700
TCR γ/δ BD Horizon BV480 CD25 PE HLA DR APC/Fire 750
CD14 Brilliant Violet 510 IgD PE/Dazzle 594
CD8 Brilliant Violet 570 CD95 PE-Cy5
CD1c Brilliant Violet 605 CD11b PerCP-Cy5.5
PD-1 Brilliant Violet 650 CD38 PerCP-eFluor® 710
CD56 Brilliant Violet 711 CD57 PE-Cy7
CD4 Brilliant Violet 750
CD28 Brilliant Violet 785
The 24-Color Panel Includes Many Highly Overlapping Dyes:
Cytek Northern Lights 24-Color Panel Violet Excited Dyes Emission Spectra
Violet Excited Dyes Emission Spectra
Cytek Northern Lights 24-Color Panel Blue Excited Dyes Emission Spectra
Blue Excited Dyes Emission Spectra
Cytek Northern Lights 24-Color Panel Red Excited Dyes Emission Spectra
Red Excited Dyes Emission Spectra

queue_play_next 24-Color Data

On the next tab, this 24-color panel is demonstrated in a healthy donor using a whole blood lyse wash sample preparation.

Aurora: 3 Lasers, 24 Colors, Unparalleled Resolution

(Click to Enlarge)

Small Particle Detection

With its onboard 100mW 405nm laser and highly sensitive violet side scatter detector, particles nearing 100nm in size can be analyzed. The Aurora opens the door to a wide variety of small particle applications.

Example: ApogeeMix

Resolution of ApogeeMix (Apogee Flow Systems), mixture of beads ranging from 1300nm to 110nm, when acquired on the Aurora. The smallest particles can be easily identified above background.

Data analyzed using FCS Express 6 by De Novo Software.

Resolution of ApogeeMix, mixture of beads ranging from 1300nm to 110nm, when acquired on the Aurora.

Autofluorescence Extraction

For those challenging applications involving highly autofluorescent particles, let the software's autofluorescence extraction ability bring new levels of resolution. Spectral cytometry has the advantage of measuring the autofluorescence spectrum of your unstained specimen and allows to extract its contribution from other fluorescent parameters. This results in better resolution of markers conjugated to dyes that heavily overlap with the cells' autofluorescence signal.

Example: PrimeFlow RNA Assay

Human U937 cells were subjected to the PrimeFlow RNA Assay, underwent a series of hybridization steps to label mRNA for HMBS, a low expressed gene (~10 copies/cell), with Alexa Fluor® 488. The sample was run on the Aurora and analyzed using SpectroFlo® software with two different strategies, one with autofluorescence extraction and one without.

Spectrum plots of unstained and Alexa Fluor 488 stained cells acquired from Northern Lights.
Spectrum plots of unstained and Alexa Fluor 488 stained cells acquired from the Aurora. Note that the two spectra heavily overlap.
Northern Lights plots with clearly identified populations.
Unstained cells were mixed with cells stained for HMBS RNA. The fluorescence signal of the mixed sample was measured. Due to the high autofluorescence, separation of negative and positive signals was marginal (upper histogram). Autofluorescence extraction greatly improved the resolution of the two cell populations (lower histogram).

PrimeFlow is a trademark of Thermo Fisher Scientific.

Fluorescent Proteins and Challenging Dye Combinations

The detection of some fluorescent protein or fluorochrome combinations by conventional flow cytometry presents a challenge due to high amounts of spectral overlap (Figure 1, 4). The Aurora addresses this challenge by using differences in full emission spectra signatures across all lasers to clearly resolve these combinations, even if the populations are co-expressed (Figures 2, 3, 5, and 6).

Example 1: APC and Alexa Fluor 647
Spectrum plots from a conventional spectrum viewer.
Figure 1: Spectrum plots from a conventional spectrum viewer shows heavy overlap between APC and Alexa Fluor 647.
Spectrum plots from the Aurora.
Figure 2: Spectrum plots from a four laser Aurora show distinct signatures for APC and Alexa Fluor 647.
Northern Lights plots with clearly identified populations.
Figure 3: Whole blood from a healthy donor was stained, lysed, washed, and analysed on a four laser Aurora system. Subsets of NK and NK T cells that co-express CD56 Alexa Fluor 647 and CD8 APC were easily identified. For comparison, blood from the same donor was stained with CD56 PE and CD8 APC and yielded similar percentages of NK and NK T cells, demonstrating that APC and Alexa Fluor 647 combined did not impact results.

Example 2: BFP, GFP, and mCherry
Spectrum plots from a conventional spectrum viewer.
Figure 4: Spectrum plots from a conventional spectrum viewer.
Spectrum plots from Northern Lights.
Figure 5: Spectrum plots from a four laser Aurora show distinct signatures for BFP, GFP and mCherry.
Northern Lights plots with clearly identified populations.
Figure 6: AB2.2 mouse Embryonic Stem Cells were genetically modified to stably express BFP, GFP and mCherry under the control of different fate marker promoters. The stable cell line generated was then cultured under differentiation conditions, harvested, and analysed on the Aurora to assess the expression of fluorescent proteins. Autofluorescence extraction was used to enhance results. Sample courtesy from Luigi Russo, Hannah L. Sladitschek and Pierre Neveu, Cell Biology & Biophysics, Neveu group, EMBL.

Why Choose Aurora?


Cytek Aurora Competitor Top 13 Color Cytometer Competitor 28 Color Cytometer Competitor 30+ Color Cytometer Competitor Spectral Cytometer
Maximum number of detectors per laser 16 5 10 10 32
Spatially separated lasers Yes Yes Yes Yes No
20-color assay sensitivity Excellent N/A Average Average Sub-optimal
Supported fluorescent tags All existing dyes Limited by optical filters provided Limited by optical filters provided Limited by optical filters provided Limited: red and violet lasers are co-linear
Detection emission wavelength range 400-900nm 400-800nm 400-800nm 400-800nm 500-800nm, 430nm, 460nm
Special fluorochromes needed for 20 color assay None N/A None, but limited fluorochrome choices Yes, but limited to exclusive fluorochromes None, but limited fluorochrome choices
Ability to test new dyes excited by supported lasers Yes Requires new filters Requires new filters Requires new filters Yes
Instrument setup to optimize sensitivity Automatic Manual Manual Manual Manual
Unmixing capability for overlapping dyes Yes No No No Yes
Able to remove cell autofluorescence Yes No No No Yes
Footprint Small Very Small Medium Large Medium

Stain Index Comparison

(Click to Enlarge)

Publications

Title Publication Abstract Published Download
Loss-of-function mutations in ATP6AP1 and ATP6AP2 in granular cell tumors Nature Communications Granular cell tumors (GCTs) are rare tumors that can arise in multiple anatomical locations, and are characterized by abundant intracytoplasmic granules. The genetic drivers of GCTs are currently unknown. Here, we apply whole-exome sequencing and targeted sequencing analysis to reveal mutually exclusive, clonal, inactivating somatic mutations in the endosomal pH regulators ATP6AP1 or ATP6AP2 in 72% of GCTs. Silencing of these genes in vitro results in impaired vesicle acidification, redistribution of endosomal compartments, and accumulation of intracytoplasmic granules, recapitulating the cardinal phenotypic characteristics of GCTs and providing a novel genotypic–phenotypic correlation. In addition, depletion of ATP6AP1 or ATP6AP2 results in the acquisition of oncogenic properties. Our results demonstrate that inactivating mutations of ATP6AP1 and ATP6AP2 are likely oncogenic drivers of GCTs and underpin the genesis of the intracytoplasmic granules that characterize them, providing a genetic link between endosomal pH regulation and tumorigenesis. August 30, 2018 Download
CBLB Constrains Inactivated Vaccine–Induced CD8+ T Cell Responses and Immunity against Lethal Fungal Pneumonia The Journal of Immunology Fungal infections in CD4+ T cell immunocompromised patients have risen sharply in recent years. Although vaccines offer a rational avenue to prevent infections, there are no licensed fungal vaccines available. Inactivated vaccines are safer but less efficacious and require adjuvants that may undesirably bias toward poor protective immune responses. We hypothesized that reducing the TCR signaling threshold could potentiate antifungal CD8+ T cell responses and immunity to inactivated vaccine in the absence of CD4+ T cells. In this study, we show that CBLB, a negative regulator of TCR signaling, suppresses CD8+ T cells in response to inactivated fungal vaccination in a mouse model of CD4+ T cell lymphopenia. Conversely, Cblb deficiency enhanced both the type 1 (e.g., IFN-γ) and type 17 (IL-17A) CD8+ T cell responses to inactivated fungal vaccines and augmented vaccine immunity to lethal fungal pneumonia. Furthermore, we show that immunization with live or inactivated vaccine yeast did not cause detectable pathologic condition in Cblb−/− mice. Augmented CD8+ T cell responses in the absence of CBLB also did not lead to terminal differentiation or adversely affect the expression of transcription factors T-bet, Eomes, and RORγt. Additionally, our adoptive transfer experiments showed that CBLB impedes the effector CD8+ T cell responses in a cell-intrinsic manner. Finally, we showed that ablation of Cblb overcomes the requirement of HIF-1α for expansion of CD8+ T cells upon vaccination. Thus, adjuvants that target CBLB may augment inactivated vaccines and immunity against systemic fungal infections in vulnerable patients. July 27, 2018 Download