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Cytek Northern Lights

Overview
Specifications
Software
Plate Loader
Performance Data

Overview


A new flow cytometry system that shifts the paradigm in what scientists expect to see in performance from an affordable three laser system.

Cytek Northern Lights incorporates the same groundbreaking technologies as its older sister, the Cytek Aurora. Like Aurora, its optical design and unmixing algorithm give scientists remarkable flexibility, enabling the use of a wide array of new fluorochrome combinations without reconfiguring the 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 sits in a sweet spot for scientists that have budgeted for a one to three laser system, but desire the ability to run panels of higher 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 what you should expect from an affordable cytometer and has delivered an instrument that brings the benefits of full spectrum cytometry to more scientists.


High Value

Upgradeable from one laser and nine colors to three lasers and 24 colors, there is a Northern Lights configuration to fit your needs.


Remarkable Sensitivity

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

Superb resolution of dim and rare populations, even in high complexity panels and high flow rates.


Easy, Flexible, and Intuitive

One configuration for all assays - no need to change optical filters.

Use any commercially available fluorochrome excited by the onboard lasers.

Intuitive software with familiar workflow.


Low Cost of Ownership

Fewer lasers to run more colors.

Low maintenance lasers, more fluorochrome choice, one configuration, and up to 24 colors per sample equates to greater cost savings and less setup time between experiments.

Specifications

Optics

Excitation Optics

Optical Platform

Northern Lights contains a fixed optical assembly configured with one to three spatially separated laser beams. Laser delays are automatically adjusted during instrument QC.

Lasers

One laser configuration: 488nm: 50mW
Two laser configuration: 488nm: 50mW, 640nm: 80mW
Three laser configuration: 405nm: 100mW, 488nm: 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.

SSC: Two high-performance semiconductor detectors with 405nm and 488nm bandpass filter. Note: 405nm side scatter applies to three laser configurations only.

Fluorescence Detectors

Proprietary high sensitivity Coarse Wavelength Division Multiplexing (CWDM) semiconductor array per laser enabling more efficient spectrum capture in the 420-830nm range. No filter changes required for any fluorochrome excited by the 405nm, 488nm, and 640nm lasers.

Standard Optical Configuration

Violet detector module (only available in three laser configurations): 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.

Fluidics

Sample Flow Rates

Low: 15 µL/min, Medium: 30 µL/min, High: 60 µL/min, Plate high-throughput mode: 100 µL/min

Fluidic Modes

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.

Sample Dead Volume

5μL for 12x75mm tube

Plate Loader Option

96-well microtiter plate capability

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® HD 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: 61 kg
Loader weight: 13 kg

Computer Dimensions

29.1 x 9.25 x 34.4 cm

Recommended Workspace

157 x 72 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 Northern Lights Product Brochure

Click below to download the complete product brochure for the Northern Lights. 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 Northern Lights

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 Northern Lights. 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


The Power of Full Spectrum Cytometry

Full spectrum cytometry opens a wide range of new possibilities. Northern Lights allows scientists to run complex multicolor experiments with as few as one or two lasers. The unique optical system enables the use of dyes with highly overlapping peak emissions without sacrificing resolution, translating to more flexibility in dye choice. Only one configuration is used for all applications, saving time in experimental setup, and minimizing the chance for experimental error. The following experiments are examples of what is possible with Northern Lights:

9-Color Blue Laser Panel

Peripheral blood mononuclear cells (PBMCs) were thawed, stained, washed, and analyzed on the Northern Lights (two laser configuraton). In this nine color blue laser excitable dyes panel, monocytes and several CD4 T cell and CD8 T cell subsets were easily identified. Markers and fluorochromes used in this assay are summarized in the table below.

Specificity Fluorochrome
CD45RA Alexa Fluor® 488
CD3 Alexa Fluor® 532
CD27 PE
CCR7 PE/Dazzle 594
CD4 PE-Cy5
CD45 PerCP-Cy5.5
CD8 BD Horizon BB700
CD14 PerCP-eFluor® 710
CD127 PE-Cy7
Cytek Northern Lights 9-Color Panel Dyes Emission Spectra
9-Color Panel Dyes Emission Spectra
Cytek Northern Lights 9-Color Blue Laser Panel

24 Colors With Three Lasers... Is it Possible?

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
24-Color Data:

The 24-color panel is demonstrated in a healthy donor using a whole blood lyse wash sample preparation.

Northern Lights: 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. Northern Lights 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 Northern Lights. 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 Northern Lights.

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 Northern Lights 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 Northern Lights. 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 (Figures 1, 4). Northern Lights 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: GFP and YFP
Spectrum plots from a conventional spectrum viewer.
Figure 1: Spectrum plots from a conventional spectrum viewer shows heavy overlap between GFP and YFP.
Spectrum plots from Northern Lights.
Figure 2: Spectrum plots from Northern Lights show distinct signatures across three lasers.
Northern Lights plots with clearly identified populations.
Figure 3: Sp2/0 cells were transfected with GFP, YFP, CFP and/or DsRed (alone or in combination) and run on Northern Lights (plots are gated on FSC vs SSC). Each population is clearly identified.

Example 2: Qdot 705 and BV711
Spectrum plots from a conventional spectrum viewer.
Figure 4: Spectrum plots from a conventional spectrum viewer shows heavy overlap between Qdot 705 and BV711.
Spectrum plots from Northern Lights.
Figure 5: Spectrum plots from Northern Lights show distinct signatures for Qdot 705 and BV711.
Northern Lights plots with clearly identified populations.
Figure 6: Normal human whole blood was stained, lysed, washed, and analyzed on Northern Lights. Subsets of NK and NK T cells that co-express CD56 BV711 and CD8 Qdot 705 were easily identified.