High Content Screening
FUJIFILM Wako Automation, North American and European distributor of:
Yokogawa CV8000 Confocal Imaging Platform
Yokogawa has been a leader in the world of imaging for over 20 years since the introduction of the Yokogawa CSU-10. This confocal spinning disk unit revolutionized the world of microscopy by introducing a technology that enabled confocal imaging of live cells. Previous to the CSU-10’s introduction confocal laser scanners would cause so much bleaching or photoxicity they were unusable for live cell assays. Standard confocal spinning disks, in use today by most competitors, block so much excitation light that the data received is generally useless. Yokogawa’s introduction of the CSU-10 microlens enhanced confocal spinning disk was truly revolutionary. Yokogawa uses two stacked spinning disks. One disk contains thousands of pinholes that make confocal imaging possible and the other contains an equal number of microlenses that focus excitation light through each pinhole significantly boosting signal compared to competitors spinning disks lacking the microlens enhancement. This is why microscopists worldwide all know the Yokogawa name. Their imaging systems provide expanded flexibility by enabling confocal and live cell assays with sufficient signal and a biologically relevant imaging period. Explore confocal imaging of live cells in fast time-lapse mode at up to 49fps or long-term confocal time lapse imaging of cells over multiple days. Confocal imaging on Yokogawa systems is the standard, rather than just an add-on option.
Check out Yokogawa’s line of High Content Imaging and Analysis products that utilize the CSU-W1 and Hamamatsu’s ORCA Flash 4.0 V3 sCMOS cameras. One system with the latest, most powerful confocal spinning disk combined with the most sensitive sCMOS camera available today. Follow up with High Content Analysis using Yokogawa’s Cell Pathfinder (CPf) analysis software and utilize Fujifilm Wako Automation’s Wako Software Suite (WSS) designed to quickly search for objects of interest in low resolution mode and quickly re-image in high resolution. Plus, WSS does so much more. Read below.
The Yokogawa CV8000 is the fastest, highest resolution, and most flexible High Content confocal solution for today’s scientist. The CV8000 is also the ideal imaging system for live cell multiday assays, tissue section imaging on slides, performing high speed kinetic imaging, imaging Cell Painting Assays, and spheroid/organoid imaging. Yokogawa’s name is synonymous with confocal microscopy since introducing the microlens enhanced confocal spinning disk system almost 25 years ago. Yokogawa has continuously improved its systems culminating in the unrivaled CV8000 which combines the highest resolution spinning disk available today, the CSU-W1, with the highest sensitivity sCMOS camera, the Hamamatsu ORCA Flash 4.0 V3.
The CV8000’s flexible design enables scientists to visualize small objects more clearly because of the greater resolution provided by the latest Yokogawa confocal unit and the amazing quantum efficiency of the best sCMOS camera on the market. In addition, resolution and light gathering can be further improved by purchasing Yokogawa’s optional 20x, 40x, and 60x water immersion objectives.
Imaging spheroids, organoids or large objects and need to improve speed and image quality? Purchase the optional dual 25µm and 50µm confocal pinhole option to further improve resolution beyond anything else available for HCS. The 20x 1.0NA water objective combined with the 25µm pinhole enables high quality spheroid images, reduces or eliminates the need to stitch dozens of images together, and provides higher quality results and a speedier workflow.
While some competitors make compromises in terms of flexibility or try to reduce costs by using a less sensitive camera, low powered lasers or ineffective spinning disks, Yokogawa makes no compromises, providing the fastest, most sensitive, and most flexible high content imaging system on the market.
Microscopists have trusted the Yokogawa name since they developed the first microlens enhanced Nipkow spinning disk almost 25 years ago. Microscopy is at the heart of High Content Imaging. Trust the brand that microscopists have always known and use.
- The CV8000 provides up to four large format Hamamatsu Orca Flash 4.0 V3 sCMOS cameras with simultaneous acquisition in up to four channels for unrivaled speed, but the most amazing part of the CV8000 is that it is so fast it beats most systems under most conditions even when using sequential imaging. Sequential imaging eliminates 100% of crosstalk that can occur from simultaneous imaging.
- Yokogawa’s 60x, 40x, and 20x high NA water immersion objectives capture more light and provide high image resolution, which is especially helpful for spheroid and tissue imaging.
- Image in fluorescence, bright field, true phase contrast, or digital phase contrast modes to see which mode meets your imaging needs.
- The sCMOS sensor provides almost 4x larger field of view compared to traditional CCD sensors allowing scientists to image more cells per field.
- The CV8000 uses Hamamatsu’s cutting-edge ORCA flash 4.0 V3 sCMOS sensor providing the highest quantum efficiency of any sCMOS on the market today. This high sensitivity camera enables CV8000 users to see objects not possible on other microscopes. Increasing exposure times also increases background. Nothing can make up for a camera with a low quantum efficiency.
- Flexibility is key with the CV8000. The 25µm pinhole option provides another level of resolution and enables the use of lower magnification objectives without sacrificing image quality. Use the 20x water objective with the new 25µm pinhole option and make image stitching a thing of the past.
- Yokogawa invented the microlens enhanced confocal spinning disk to enable confocal imaging of live samples while significantly reducing the risks of phototoxicity and bleaching. The CV8000 provides temperature, humidity, and CO2 control to enable true live cell multiday imaging.
- The optional FRET mode further expands your assays choices.
- Yokogawa’s in house developed laser-based autofocus system enables imaging of 384 wells plates with zero autofocus errors virtually every time.
- HCS instrumentation can produce a lot of data. Yokogawa understands minimizing the amount of data saved is important to many customers. This is why the CV8000 has the ability to save only MIP (Maximum Image Projection) images. Users may take many z slice images, but may plan to analyze them as MIPs in 2D, therefore the individual z slice images are not needed. Imagine how much space this can save. If a customer images 20 z slices in 4 colors per field, but only needs a MIP, the CV8000 will only save the 4 MIPs (one for each color) rather than 80 individual z slice images plus 4 MIPs. Remember, this is for every field, of every well, of every plate. This ends up saving a massive amount of hard drive space. In addition, Wako’s Software Suite has a mode that enables all CV8000 images to be compressed using a choice of lossless compression formats (LZW, LZW:2, ZIP). This can reduce file size by up to 40%.
Talk to an imaging specialist today and design a CV8000 with the options that best match your research needs.
Yokogawa’s Cell Pathfinder (CPf) Image Analysis Software makes converting images into numbers easy
- Use simple built-in templates for common analysis routines to quickly build a protocol or switch to the advanced mode for more sophisticated analysis.
- Use Machine Learning to teach CPf which objects you’re interested in.
- Manually draw regions of interest or manually select specific objects for precision control.
- CPf uses a simple workflow design enabling scientists to intuitively build an analysis protocol.
- CPf provides dozens of output features including texture morphology that can be used to build deep learning models as well as 3D output features such as volume and surface area.
- Export your results into csv files and store along with standard Tiff images in your preferred database or import into your favorite data analytics system.
Wako Software Suite
- ScreenSentry™ – Automatically perform a quick plate QC before starting a multi-hour screen. Screen smarter. Screen faster.
- SearchFirst™ – Quickly scan your plates using a low power objective then automatically rescan objects of interest in high resolution
- Run Method – Enhance the CV8000’s capabilities by enabling additional options available in Wako’s Run Method module
|Detection Methods||Bright Field, True Phase Contrast, Digital Phase Contrast, Confocal|
|Lasers||405nm, 488nm, 561nm, 640nm|
|Bright Field Light Source||Xenon Arc Lamp|
|FRET Option||Additional 445 nm Laser|
(Up to 6 per camera)
|445/45, 525/50, 600/37, 676/29, Wide band 447-522,
Wide band 488-568, 488/32 (CFP), 549/30 (YFP), 589/18, 618/26, 625/15
(up to 6)
Dry: 2x 0.8NA, 4x 0.16NA, 10x 0.4NA, 20x 0.75NA, 20x LW 0.45NA, 20x Phase 0.4NA, 40x 0.95NA
Water: 20x 1.0NA, 40x 1.0NA, 60 1.2NA
(Up to 4)
|Hamamatsu ORCA Flash 4.0 V3, 2200×2000 pixels, 6.5µm pixel|
|Confocal Spinning Disk||Yokogawa CSU-W1 Nipkow microlens enhanced dual spinning disk,
50µm pinhole or dual 50µm/25µm pinhole option
|Liquid Handling||100µl or 30µl disposable tips|
|Environmental Controls||Active Humidification, Temperature, CO2, Hypoxic Conditions (N2)|
Yokogawa Cell Pathfinder (CPf)
Designed with both screeners and microscopists in mind, CPf’s utilizes simple application templates for standard assays such as Cell Cycle and Translocation, a simplified basic mode for users who are new to high content analysis, and an advanced mode with over 100 selections to optimize object identification. All modes are designed with a flexible and straightforward workflow-based interface that makes the entire process simple but powerful.
Power and Flexibility:
- Live Cell Long Term Time Lapse (Image a plate for 3+ days while maintaining healthy cells utilizing the CV8000’s CO2, Temperature and Active Humidification)
- CPf’s Cell Tracking module provides 9 parameters users can adjust to ensure the same object is consistently tracked frame to frame
- Track cell division. Output links mother cells to daughter cells
- Track speed, distance moved, Linearity of Motion
- Live Cell High Speed Time Lapse (Image at almost 50fps and add inhibitors/activators in real time using the CV8000’s built in disposable tip pipettor)
- CPf’s Time in Same Region model creates a maximum image projection across time. This enables CPf to create a single fixed area to measure changes over the course of a high speed time lapse assay.
- Run Ca+ assays, cardiomyocyte assays, and more
- 3D Imaging
- Build 3D models of your cells, pull out more information, and expand the knowledge of your biological system
- Report volume, surface area, and count objects within spheroids, organoids, or PDX tumor models.
- Analyze in 3D or convert to maximum, minimum, average, or sum image projection
- Quickly capture rare events, improve imaging speed, and locate spheroids
- First Scan using low powered objectives then acquire objects of interest at high resolution
- Machine Learning and Manual Controls
- Utilize the machine learning module to teach CPf how to distinguish objects of interest from each other and from background.
- Extremely powerful tool for ill-defined objects
- Manually trace objects of interest or regions of interest to ensure only those objects important to the experiment are being captured.
- Texture Analysis
- Utilize texture analysis to identify differences between cell states that may not be apparent when using standard intensity-based output.
- Report Peak, Valley, Hole, Edge, Ridge, and Saddle textures
- Control pixel distance, test and compare results
- Separate cell with distinct characteristics into populations using the gating module
Wako Software Suite
Wako Software Suite (WSS) is software designed by FUJIFILM Wako Automation that enhances the functionality of the Yokogawa CV8000. Wako takes a pragmatic approach to imaging. Often more efficient workflows lead to much greater returns than can be accomplished through hardware alone. WSS was designed not only to provide applications that significantly improve the efficiency and flexibility of the CV8000, but it is used to rapidly add new features requested by customers. The best part of WSS is that it comes totally free with the purchase of a Yokogawa CV8000.
One Powerful Suite of Software Turns Your Idea Into Reality
They say imitation is the best form of flattery. Wako invented this powerful module over 5 years ago, and though others have tried to copy the idea, none have come close to the flexibility and power of SearchFirst™. SearchFirst™ enables customers to automatically identify objects of interest and then perform a detailed scan on those objects. While competitors have copied the basics of this function to enable the rapid location of spheroids so that many images can be taken for complex modeling, SearchFirst™ does so much more.
SearchFirst™ is incredibly flexible. Customer have full control of the pre-scan method used to quickly locate objects of interest. For example, any objective, binning, number of fields or wells, or combination of colors can be used. The same goes for detailed imaging once your object of interest is located. Make your high resolution method as complex or simple as you wish. Analysis is just as flexible as method design. Analysis protocols used to identify objects of interest can be created using a number of analysis software platforms including Yokogawa’s Cell Pathfinder, KNIME, ImageJ, CellProfiler, MatLab, and Python scripts. Some of the major features of SearchFirst™ are listed below:
- Center on Objects – Identify objects of interest, center on objects, and re-image. Add additional fields and stitch if the object is too large to fit into a single field. This method is best used for large objects present in small numbers, such as spheroids.
- Center, but Avoid Oversampling – Identify objects of interest, center on object, but do not re-image objects of interest located close together as the same object may show up in multiple images leading to an oversampling of data that will result in data bias. This method is ideal for imaging non-confluent cells or for imaging a relatively small population of cells. Most cells that meet the selection criteria will be imaged, and many will be centered, but some may be left out in order to avoid oversampling of data.
- Grid Option – This option turns your first pass field of view into a grid of second pass fields of view. Only second pass fields that contain hits are imaged. This method is good for imaging small to large populations of cells. All cells meeting the selection criteria will be imaged, but they will not be centered in the field of view.
- Image Fields With Most Hits – This option lets a user enter the maximum number of fields to image. SearchFirst™ will sort the fields from most to least by the number of cells that meet the selection criteria. Only a limited number of fields that contain the most cells are imaged.
- Cell Count – This option lets users enter a cell count number. SearchFirst™ will sort the fields by number of cells meeting the selection criteria from most to least and image the least number of fields required to hit the cell count target.
At Wako we believe the best way to improve throughput is to improve efficiency. Screen Sentry™ enables users to quickly scan and QC a plate before a potentially time-consuming imaging routine begins. Test positive/negative controls for window, Z factor, cell number, or apoptosis. If a plate fails QC, it’s automatically ejected and not imaged. If a plate passes QC the full imaging method proceeds. Analysis software platforms compatible with Screen Sentry are KNIME, ImageJ, CellProfiler, MatLab, and Python scripts.
This Module enables the WSS to control the CV8000, rather than using the CV8000 user interface. Since WSS is controlling the CV8000, numerous user requested options can be enabled. Below is a sample of user requested features that have been enabled by WSS:
- Turn off laser power immediately after a plate is finished
- Automatically convert CV8000 raw tiff images into lossless compressed tiffs
- Immediately auto-launch analysis methods creating in Matlab, python, KNIME, CellProfiler, or ImageJ
- Strip or modify specific metadata from CV8000 files that may cause issues with your workflow
- Modify the default CV8000 image naming scheme
- Manually fine tune the estimated read time the CV8000 sends to automated systems to improve the scheduling efficiency of your fully integrated CV8000
In addition, to the functions listed above Wako has created numerous custom executables to help our customers manipulate metadata so that complex assay designs can fit seamlessly into our customer’s analysis workflow. Generally, custom software requests only take days to weeks to implement. When was the last time you requested a new feature and received it in days, instead of years? This is the power of Wako’s team of software engineers dedicated to industry leading customer satisfaction.