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.
51 channels of detection over the full emission spectra.
Up to 24 colors demonstrated including fluorochrome with emission spectra in close proximity to each other.
Sensitivity redefined using state-of-the-art optics and low-noise electronics.
No changing optical filters for any fluorochrome.
Use any commercially available fluorochrome excited by the onboard lasers.
A powerful, high value system that is accessible to a wide range of users.
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.
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.
Three laser configuration: 405nm: 100mW, 488nm: 50mW, 640nm: 80mW
Four laser configuration: 405nm: 100mW, 488nm: 50mW, 561nm: 50mW, 640nm: 80mW
Flat-Top laser beam profile with narrow vertical beam height optimized for small particle detection.
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.
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.
Sample Flow Rates
Low: 15 µL/min, Medium: 30 µL/min, High: 60 µL/min, Plate high-throughput mode: 100 µL/min
Long clean, SIT flush, Purge filter, 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.
Volumetric measurement during sample recording enables calculation of counts per µL for any gated population.
Plate Loader Option
96-well microtiter plate capability
Throughput time 35 minutes at High Throughput mode sampling 7 µL/well
Plate stage temperature: 4-30°C
Plate Loader Carryover
Default mode: ≤0.3%, Low Carryover mode: ≤0.1%, High Throughput mode: ≤1%
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.
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 0.2µm beads from noise.
Data Acquisition Rate
* Three-laser system
Live unmixing during acquisition
Developed specifically to streamline assay setup, data acquisition, and file export.
Automated QC module
Raw and Unmixed FCS 3.1 files
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.
Windows® 10 Pro 64-bit
Intel® Core™ i7 processor, 3.6 GHz
500GB SSD / 1TB SATA
32” UHD 4K Monitor
Dimensions (W x D x H)
Without loader: 54 x 52 x 52 cm
With loader: 58.4 x 62 x 52 cm
Instrument weight (4 lasers): 80 kg
Loader weight: 13kg
29.1 x 9.25 x 34.4 cm
157 x 71 x 132 cm
100-240V, 50/60 Hz, 2A max
500W with all solid-state lasers
20%-85% relative non-condensing
No excessive dust or smoke
No special requirements
For Research Use Only. Not for use in diagnostic or therapeutic procedures.
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
Cytek® Aurora Technical Specs
Click below to download the technical specs for the Aurora. It contains everything you need to know about this revolutionary product.Download PDF
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.
Add or remove experiment templates, worksheet templates, fluorochrome information, QC bead information, and more.
Unmix data using controls from different experiments or apply virtual filters to your data.
For administrative controls.
Customize the software appearance. Set default plot sizes, text sizes and fonts, gate colors, print layout, statistics table options, and more.
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.
Get to Know Our Automatic Micro-Sampling System (AMS)
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.
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:
|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-Cy™5|
|CD1c||Brilliant Violet 605™||CD11b||PerCP-Cy™5.5|
|PD-1||Brilliant Violet 650™||CD38||PerCP-eFluor® 710|
|CD56||Brilliant Violet 711™||CD57||PE-Cy™7|
|CD4||Brilliant Violet 750™|
|CD28||Brilliant Violet 785™|
The 24-Color Panel Includes Many Highly Overlapping Dyes:
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.
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.
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.
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
Example 2: BFP, GFP, and mCherry
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||420-830nm||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|
Stain Index Comparison
(Click to Enlarge)
|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|
|A CD4+ T cell population expanded in lupus blood provides B cell help through interleukin-10 and succinate||Nature Medicine||Understanding the mechanisms underlying autoantibody development will accelerate therapeutic target identification in autoimmune diseases such as systemic lupus erythematosus (SLE). Follicular helper T cells (TFH cells) have long been implicated in SLE pathogenesis. Yet a fraction of autoantibodies in individuals with SLE are unmutated, supporting that autoreactive B cells also differentiate outside germinal centers. Here, we describe a CXCR5−CXCR3+ programmed death 1 (PD1)hiCD4+ helper T cell population distinct from TFH cells and expanded in both SLE blood and the tubulointerstitial areas of individuals with proliferative lupus nephritis. These cells produce interleukin-10 (IL-10) and accumulate mitochondrial reactive oxygen species as the result of reverse electron transport fueled by succinate. Furthermore, they provide B cell help, independently of IL-21, through IL-10 and succinate. Similar cells are generated in vitro upon priming naive CD4+ T cells with plasmacytoid dendritic cells activated with oxidized mitochondrial DNA, a distinct class of interferogenic toll-like receptor 9 ligand. Targeting this pathway might blunt the initiation and/or perpetuation of extrafollicular humoral responses in SLE.||November 26, 2018||Download|
|Cell by cell immuno- and cancer marker profiling for non-small cell lung cancer (NSCLC) tissue sample using non-enzymatic tissue dissociation and high-parameter flow cytometry||American Association for Cancer Research||Non-small cell lung cancer (NSCLC) is the most common type of lung cancer, accounting for 80-85% cases of lung cancer. It has poor prognosis as most NSCLC patients are at advanced stage when diagnosed. The development of immunotherapy blocking the PD-1/PD-L1 ligand pathway, in recent years, has shed light on new treatment for lung cancer. An in-depth understanding of the interaction between immune system and tumor will help to identify novel markers for lung cancer treatment.
In this study, we performed a comprehensive study on 10 NSCLC patient tissue samples with paired blood samples for circulating tumor cells (CTCs). The solid tissue biopsy samples were dissociated into single cells by non-enzymatic tissue homogenization (IncellDx IncellPREPTM). The single cell suspensions were stained simultaneously with multiple (>20) immune check point markers (including PD-1*, PD-L1*, TIM-3*, LAG-3*, and CTLA-4*), cancer markers (such as EGFR* and ALK* fusion protein), and a cell cycle dye. The samples were interrogated on a novel, innovative, high parameter, spectral flow cytometer Cytek AuroraTM. Markers on subsets of immune cell and cancer cell populations were investigated.
Our results showed the association of high levels of immune check point marker and cancer marker expressions with the high level of aneuploidy (indicated by DNA index >1.05), and the aggressiveness of the cancer (indicated by the number of CTCs). High numbers of CTCs were associated with aneuploidy, increased Post G0-G1%, and high expression of LAG-3, TIM-3, and PD-1.
Multi-parametric flow cytometry allows simultaneous profiling of multiple immune and cancer markers on cancer samples at the single cell level. The knowledge acquired from these studies will enhance our understanding of cancer immune system, cancer cells, and the interaction between immune system and the cancer. It will potentially transform patient diagnosis, disease monitoring, and drug discovery. Our study demonstrated a powerful method to study solid tumors that may provide important information for successful precision cancer immunotherapy.
*PD-1: programmed death-1; *PD-L1: programmed death-ligand 1; *TIM-3: mucin domain-3-containing molecule-3; *LAG-3: lymphocyte-activation gene-3; *CTLA-4: cytotoxic T-lymphocyte antigen-4; *EGFR: epidermal growth factor receptor; *ALK: anaplastic lymphoma kinase
|July 1, 2018||Download|
|Detection of Succinate by Intestinal Tuft Cells Triggers a Type 2 Innate Immune Circuit||Immunity||In the small intestine, type 2 responses are regulated by a signaling circuit that involves tuft cells and group 2 innate lymphoid cells (ILC2s). Here, we identified the microbial metabolite succinate as an activating ligand for small intestinal (SI) tuft cells. Sequencing analyses of tuft cells isolated from the small intestine, gall bladder, colon, thymus, and trachea revealed that expression of tuft cell chemosensory receptors is tissue specific. SI tuft cells expressed the succinate receptor (SUCNR1), and providing succinate in drinking water was sufficient to induce a multifaceted type 2 immune response via the tuft-ILC2 circuit. The helminth Nippostrongylus brasiliensis and a tritrichomonad protist both secreted succinate as a metabolite. In vivo sensing of the tritrichomonad required SUCNR1, whereas N. brasiliensis was SUCNR1 independent. These findings define a paradigm wherein tuft cells monitor microbial metabolites to initiate type 2 immunity and suggest the existence of other sensing pathways triggering the response to helminths.||July 17, 2018||Download|
|Histone acetyltransferase CBP is critical for conventional effector and memory T-cell differentiation in mice||Journal of Biological Chemistry||Compared with naïve T cells, memory CD8+ T cells have a transcriptional landscape and proteome that is optimized to generate a more rapid and robust response to secondary infection. Additionally, rewired kinase signal transduction pathways likely contribute to the superior recall response of memory CD8+ T cells, but this idea has not be experimentally confirmed. Herein, we utilized an MS approach to identify proteins that are phosphorylated on tyrosine residues in response to Listeria-induced T-cell receptor (TCR) stimulation in both naïve and memory CD8+ T cells from mice and separated by fluorescence- and flow cytometry–based cell sorting. This analysis identified substantial differences in tyrosine kinase signaling networks between naïve and memory CD8+ T cells. We also observed that an important axis in memory CD8+ T cells couples Janus kinase 2 (JAK2) hyperactivation to the phosphorylation of CREB-binding protein (CBP). Functionally, JAK2-catalyzed phosphorylation enabled CBP to bind with higher affinity to acetylated histone peptides, indicating a potential epigenetic mechanism that could contribute to rapid initiation of transcriptional programs in memory CD8+ T cells. Moreover, we found that CBP itself is essential for conventional effector and memory CD8+ T-cell formation. These results indicate how signaling pathways are altered to promote CD8+ memory cell formation and rapid responses to and protection from repeat infections.||December 20, 2018||Download|
|The role of systems biology approaches in determining molecular signatures for the development of more effective vaccines||Expert Review of Vaccines||Introduction: Emerging infectious diseases are a major threat to public health, and while vaccines have proven to be one of the most effective preventive measures for infectious diseases, we still do not have safe and effective vaccines against many human pathogens, and emerging diseases continually pose new threats. The purpose of this review is to discuss how the creation of vaccines for these new threats has been hindered by limitations in the current approach to vaccine development. Recent advances in high-throughput technologies have enabled scientists to apply systems biology approaches to collect and integrate increasingly large datasets that capture comprehensive biological changes induced by vaccines, and then decipher the complex immune response to those vaccines.
Areas covered: This review covers advances in these technologies and recent publications that describe systems biology approaches to understanding vaccine immune responses and to understanding the rational design of new vaccine candidates.
Expert opinion: Systems biology approaches to vaccine development provide novel information regarding both the immune response and the underlying mechanisms and can inform vaccine development.
|February 11, 2019||Download|
|Direct Inhibition of MmpL3 by Novel Antitubercular Compounds||ACS Infectious Diseases||MmpL3, an essential transporter involved in the export of mycolic acids, is the proposed target of a number of antimycobacterial inhibitors under development. Whether MmpL3 serves as the direct target of these compounds, however, has been called into question after the discovery that some of them dissipated the proton motive force from which MmpL transporters derive their energy. Using a combination of in vitro and whole-cell-based approaches, we here provide evidence that five structurally distinct MmpL3 inhibitor series, three of which impact proton motive force in Mycobacterium tuberculosis, directly interact with MmpL3. Medium- to high-throughput assays based on these approaches were developed to facilitate the future screening and optimization of MmpL3 inhibitors. The promiscuity of MmpL3 as a drug target and the mechanisms through which missense mutations located in a transmembrane region of this transporter may confer cross-resistance to a variety of chemical scaffolds are discussed in light of the exquisite vulnerability of MmpL3, its apparent mechanisms of interaction with inhibitors, and evidence of conformational changes induced both by the inhibitors and one of the most commonly identified resistance mutations in MmpL3.||March 18, 2019||Download|