CV8000 High-Throughput System

High content analysis software CellPathfinder is updated.
Deep Learning option is released.:About Deep learning

The CV8000 features a cell incubator with an improved airtight design that facilitates the observation of cell behavior over long periods of time. In addition, the CV8000 comes with CellPathfinder, a new program that can analyze images of unlabeled cells and 3D images of samples. With these features, the CV8000 improves the efficiency of drug discovery research and biomedical research on leading-edge subjects such as iPS and ES cells.

CV8000 System Highlights

Excitation laser wavelength 405nm、445nm、488nm、561nm、640nm
Illumination source Laser
Objective lens 2x to 60x
(Dry, Phase contrast, Water immersion, Long working distance)
Camera High-sensitivity sCMOS camera (up to 4 units)
Autofocus Laser-based mode, image-based mode
Software CellPathfinder、CellLibrarian

 

Introduction

Valley of death

Within the drug development market, demands on high content analysis systems for drug efficacy evaluation are increasing in accordance with the needs for cell-based assay and phenotypic screening. In order to increase screening efficiency, devices with higher speeds (higher throughput) are required.
On the other hand, in order to bridge the “valley of death” of the drug development process, the quality of screening hits must be increased.
This requires the construction of more complex evaluation systems that utilize multifaceted parameters via 3D cultivation systems, live-cell imaging and higher detail image analysis.
In current drug development research, determining how to implement throughput screening and complex evaluation system screening in parallel is an important issue.

Solution

The CellVoyager CV8000 is a high-end, high content analysis system that solves this contradictory screening challenge.
Through the combination of a proprietary Yokogawa high speed confocal scanner, water immersion lens, up to four high field-of-vision cameras, a microscopic stage with cell cultivation environment, and an integrated robotic pipetter, we have realized not only high throughput, high-resolution imaging, but also phenotypic screening via a more complex evaluation system.
In addition, our specialized analysis software, CellPathfinder, uses deep leaning and machine learning to recognize target objects with high accuracy, supporting you from image analysis to results display using graphs.

The Yokogawa advantage

  • Confocal scaner unit
  • Live/kinetic experiment compatible
  • High throughput
  • Reliable, proven technology

Voyage to unknown worlds

Real time confocal, label-free imaging

Long-term liv e c ell imaging

Long-term livecell imaging

Stage incubator included as standard. Realization of non-stop, long-duration observation (3 days +) via humidity, temperature and CO2 control.

Left:Before incubation Right:After 68 hours incubation

Kinetic assay

Kinetic assay

Drug addition during imaging is made possible by an integrated robotic pipetter with disposable tips.
Ideal for kinetic experiments involving the observation of high speed phenomena.

Left:Before incubation Right:After stimulation

Organoid / Spheroid

Organoid / Spheroid

Yokogawa’ s spinning disk confocal technology excels in imaging of samples with depth, such as 3D culture samples where clear and quick imaging is difficult, allowing for evaluation close to in-vivo quality.

Left:Original image Right:Recognition image

Label-free analysis

Label-free analysis

Recognition and analysis can be performed by taking bright field images from several Z positions and creating a CE bright field image using the included CellPathfinder analysis software. Analysis accuracy is further enhanced via the new Deep Learning option.

Left:CE Bright Field Right:Cell Recognition image

Details

State-of-the-art technology that enables you to do what you want

Observe cells as they are -Dual spinning disk confocal system-

Dual spinning disk confocal system

A Yokogawa proprietary multi-scan method utilizing approximately 1,000 laser beams on the observation region and tandem disks rotating at high speed. The disks comprise a pinhole array disk with approximately 20,000 pinholes arranged in an equal pitch spiral pattern, and a microlens array disk that focuses the excitation light laser into individual pinholes. Not only does this allow high speed imaging, but it also largely prevents phototoxicity and fluorescence photobleaching.

Deeper, clearer observation -Pinhole disk exchanger-

Pinhole disk exchanger
 
Pinhole disk exchanger

Two different types of pinhole disks (25/50μm) can be used, according to the sample. For thick samples, reducing the pinhole diameter allows for higher confocality, shaper images. For dark samples, increasing the pinhole diameter allows for brighter images.

Organoid imaging example Upper:25μm pinhole Lower:50μm pinhole

Higher throughput screening -Optical configuration-

Optical configuration

The optical system configuration can be selected according to the purpose. A single 96-well plate can be imaged in four colors in one minute by attaching four high- sensitivity wide -field sCMOS cameras. The system is also compatible with FRET and CellPainting assay.

Capturing finer structures -Original water immersion lens-

water immersion lens

Water immersion lenses excel in capturing high-resolution images of cells within a liquid. The CV8000 can be equipped wit h a 40x or 60x water immersion objective lens. Our 40x lens is a particularly unique lens capable of highly advanced spherical aberration correction, allowing for the capture of bright high-resolution images over a full wide -field. The lens water supply is also completely automated. This equipment makes high throughput screening via water submersion lens possible.

Capturing live cell movement -High-precision incubator and robot pipetter-

robot pipetter

The stage incubator features an airtight construction, managing humidity, temperature and CO2 levels. The integrated robotic pipetter conducts the following process fully automatically: tip pickup → reagent collection from the reagent plate → reagent addition to the sample plate → tip disposal. Not only can images be rapidly obtained before and after reagent instillation, but it’s also possible to add reagents to single wells multiple times, and adjust the addition speed etc., broadening the range of dynamics observation via reagent instillation.

A more live cell-friendly total HCA system

Making long-duration live -cell imaging possible -Featuring a s table built -in s tage incub ator-

HeLa cells were seeded in a 96 well plate at a density of 500 cells per well, and cultured for 24 hours. The well plate was then placed in the internal stage incubator and cell culturing was conducted for 72 hour s, and the total area (hereinafter Total Area) occupied by cells was analyzed. As a result, minimal unevenness in cell multiplication was observed across the 96-wells (excluding the four corner wells) when compared to a regular CO2 incubator.

Cell multiplication comparison with regular CO<sub>2</sub> incubator after 72hr incubation

Cell multiplication comparison with regular CO2 incubator after 72hr incubation(n=3)

  • 96 well average: 90
  • Average of outermost 36 wells: 81
  • Average of 60 wells (excl. outermost): 96

The values represent the following: CV8000 Total Area after 72hrs / Total Area at 0hrs (hereinafter Total Area ratio) / CO2 incubator Total Area ratio x 100.
(Numbers near to 100 me an that cell multiplication was approximately equal for the CV8000 and CO2 incubator.)
Cell multiplication near to that of the CO2 incubator was verified, excluding the four corner wells.

Cell multiplication curves for each well of a 96-well plate

Cell multiplication curves for each well of a 96-well plate

  • Vertical axis: Total Area
  • Horizontal axis: Time (0-72 hours)

Cell multiplication was low in the four corner wells; however, it continued in the other wells.。

Total Area ratio after cultivation start

Total Area ratio after cultivation start (24, 48 and 72 hours) (n=3)

Excluding the four corner wells, even after 72 hours, there were no large differences in cell multiplication.
The low variation in cell multiplication speed across the wells can 24 hours 48 hours 72 hours be seen.

More info Evaluation of cell-culture condition in CV8000’s internal stage incubator

System Integration

Centralized process management, from the cultivation environment, to transfer, imaging, analysis and data management.
We offer optimum systems in response to our customers’ needs.

System Integration

High Content Analysis Software CellPathfinder

The software analyzes image data captured with the CV8000, creates graphs and exports various data. Beginner and expert users alike can take full advantage of the software, thanks to an abundance of templates and flexible protocol editing capability. CE bright field and machine-learning functionalities make label-free analysis possible. The new Deep Learning option has also been added, largely improving cell recognition accuracy.

More info High Content Analysis Software CellPathfinder

Just click the menu item for analysis

Just click the menu item for analysis

Simply follow the flow displayed at the top of the screen. The analysis menu has easy-to-understand icons. Simply click the desired menu item and the protocol will load.

Fast results for immediate verification and study

Fast results for immediate verification and study

Computed numeric data can be displayed in a variety of ways. Graph plots and cell images are linked, making for easy result verification and inquiry.

Unbiased phenotype evaluation via AI

Unbiased phenotype evaluation via AI

Machine-learning also provides bias-free digitization of visually-evaluated experiments. Automatic recognition is made possible simply by clicking the shape you want the software to learn.

Label-free phenotype analysis

Label-free phenotype analysis

Eliminates the time, cost and influence on cells associated with cell labelling. Even higher precision classification is made possible through combination with deep learning.

Specifications

High-throughput Cytological Discovery System

Model CV8000
Sample format Multiple well plate (6, 12, 24, 48, 96, 384, 1536 wells), glass slide
Image mode Confocal mode: max. 4 color simultaneous recording
Bright field/phase contrast (10x, 20x for 6, 12, 24 well plates), digital phase contrast (10x, 20x)
Output data format Image data: 16bit TIFF, PNG
Numerical data: CSV, original format
Excitation wavelength 405/445/488/561/640 nm, all solid laser, max. 5 lasers 
【Option】365 nm LED
White light illumination LED
Autofocus Laser-based mode, image-based mode
Objectives Max. 6 lenses are available, automatically switchable
Dry: 2x, 4x, 10x, 20x, 40x Water immersion: 20x, 40x, 60x
hase contrast: 10x, 20x Long working distance: 20x
Confocal unit Microlens-enhanced wide-view dual Nipkow disk confocal scanner, 50 μm pinhole
【Option】 25 μm pinhole disk and auto pinhole disk exchanger
Camera sCMOS (effective pixels: 2000X2000 pixel size: 6.5 μm), max. 4 cameras
Stage incubator Temperature for incubation : 35-40℃ CO2 supply box (CO2: 5%, forced humidification)

Robot pipetter

【Option】 Disposable tip type (96tip or 384tip type)
Bar code reader 【Option】 1 or 2 dimension
Workstations Dual-monitor work station for system control, dual-monitor work station for data analysis
Analysis software High Content Analysis Software CellPathfinder
Granularity, Neurite, Nuclear morphology, Nuclear translocation, Plasma membrane translocation, Machine learning, Label-free analysis, 3D analysis, Deep Learning, etc.
Operating environment 15~30℃ 30~70%RH (no condensation)
Power supply Measurement unit:AC100-240V, 50/60Hz, 2KVA max
Workstation for system control:AC100-240V, 50/60Hz, 1.3KVA max
Workstation for data analysis:AC100-240V, 50/60Hz, 950VA max
Dimensions Measurement unit: W1,280×D895×H1,450 mm
Weight Measurement unit: 510Kg

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Resources

Overview:

The CV8000 nuclear translocation analysis software enables the analysis of changes in the localization of signal molecules that transfer between cytoplasm and nuclei, such as proteins. The following is an example of the translocation analysis of NFκB, a transcription factor.

Application Note
Application Note
Application Note
Yokogawa Technical Report
1.2 MB
Industries:
Yokogawa Technical Report
2.2 MB
Overview:

List of Selected Publications : CV8000, CV7000, CV6000

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Product Overview
Overview:

The CV8000 features a cell incubator with an improved airtight design that facilitates the observation of cell behavior over long periods of time. In addition, the CV8000 comes with CellPathfinder, a new program that can analyze images of unlabeled cells and 3D images of samples. With these features, the CV8000 improves the efficiency of drug discovery research and biomedical research on leading-edge subjects such as iPS and ES cells.

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