Pioneering Reside Cell Imaging – Yokogawa’s Influence and Improvements


Because the Nineties, Yokogawa has performed a distinguished function within the R&D of high-speed confocal microscopes for real-time imaging of dwelling cells, and in 1996, launched its first product, the CSU10 confocal scanner unit. The corporate then launched the CellVoyager high-throughput screening system, the CQ1 all-in-one picture cytometer, the CellPathfinder cell picture evaluation software program, and different superior merchandise. Yokogawa is a pioneer of reside cell imaging applied sciences and drug discovery help programs.

On this interview, NewsMedical speaks with Kevin Jan from Yokogawa about the way forward for reside cell imaging applied sciences.

The CSU10 was a landmark product for Yokogawa in 1996. How did its introduction impression the sector of reside cell imaging and Yokogawa’s place out there?

Utilizing confocal microscopy, the CSU 10 enabled reside imaging of cells and thick specimens by way of the z-axis with multipoint illumination and multipoint detection, whereas each the illumination and detection of typical optical microscopy have been planar. Additionally, utilizing a lot of the confocal microscopes on the time, it sometimes took over a second to supply a single picture. That was too gradual to watch the motion of dwelling cells in real-time.

The CSU10 supplied scanning speeds as quick as 360 frames/second with a excessive signal-to-noise (S/N) ratio. These traits allowed direct viewing of clear-cut, confocal photographs of fluorescent specimens in real-time on the eyepiece. The confocal photographs captured by the CSU10 could possibly be recorded by a video digicam as reside photographs. These allowed the research of real-time conduct and interactions amongst microorganisms. The imaging high quality allowed a technician to document a clear-cut cross-section of a whole-mount embryo. Plus, the CSU10 could possibly be mounted on a lot of the microscopes used on the time.

What have been the important thing technological and market challenges Yokogawa confronted whereas growing the CellVoyager high-throughput screening system?

The challenges have been really typical of a brand new, superior know-how. Other than our collaboration—or, as we are saying, co-innovation—companions, many within the trade didn’t perceive the know-how or the probabilities it enabled. Nevertheless, these concerned in superior drug discovery and improvement required what we known as “excessive content material evaluation” for phenotypic screening. The usage of 3D samples, corresponding to spheroids and organoids, and reside cell samples to imitate real looking mobile environments was rising as a key requirement because of cell tradition know-how improvement.

Assessing a really giant variety of compounds is time-consuming—however buying numerous high-resolution mobile photographs with quick scanning velocity is critically essential to drug screening. The high-content evaluation imagers balanced these typically conflicting necessities at a excessive degree of efficiency. The core know-how, the microlens-enhanced twin Nipkow disk in our confocal scanners and confocal spinning disc models (CSUs), makes it potential to seize high-quality photographs with remarkably quick acquisition time.

A Nipkow spinning disk containing about 20,000 pinholes and a subsidiary spinning disk containing the identical variety of microlenses to focus excitation laser gentle into every corresponding pinhole are mechanically fastened on a motor and rotated at excessive velocity. Consequently, a high-speed raster scan of the excitation lights on the specimen might be achieved.

The pinhole and microlenses are organized on every disk in a proprietary design to optimize the raster scan. Multi-beam scanning not solely will increase scanning velocity but in addition leads to considerably decrease photobleaching and phototoxicity as a result of multi-beam excitation wants solely a low degree of laser energy on the specimen to completely excite fluorescence.

Whereas there have been technical paradigms to beat, we have been readily in a position to present comparisons between high-content evaluation (HCA) and different strategies, corresponding to circulate cytometry, high-throughput screening, and conventional microscopy. Justifying the upper value took time, however it in the end hinged on serving to potential customers perceive the probabilities—and the numerous aggressive disadvantages of lagging within the fast-evolving drug discovery course of. Furthermore, early success tales supplied essential credibility.

Hela Cells

Picture of Hela Cells taken by CV8000_MitoGreen (Inexperienced) and Dapi (Blue) 60x Water Immersion Lens, 50um Pinhole. Picture Credit score: Yokogawa Life Science

How does the CQ1 and CQ3000 picture cytometer stand out within the aggressive panorama of picture cytometry, and what options make it notably suited to drug discovery?

The CQ1 and CQ3000 are each benchtop models that mix the operability of a standard circulate cytometer with the excessive picture high quality of our HCA know-how. They are often simply put in in any lab with out the necessity for a darkroom or specialised setup. Regardless of its ease of use, the CQ1 and CQ3000 provide wealthy characteristic extractions for stylish mobile picture evaluation. Its Nipkow spinning disk confocal know-how permits high-speed scanning whereas minimizing phototoxicity and photobleaching, considerably enhancing productiveness.

The CQ1 and CQ3000 allow measurements of spheroids, colonies, and tissue sections. In distinction to conventional circulate cytometry, cells aren’t faraway from the tradition dish. As well as, a high-precision stage incubator and low phototoxicity of the confocal make the evaluation of time-lapse and reside cells potential. It’s also suitable with the CellPathfinder high-content evaluation system software program.

Might you make clear the event journey of the CellPathfinder cell picture evaluation software program and its contributions to the sector of mobile evaluation?

As superior because the core, excessive content material evaluation {hardware} know-how is, realizing the mobile imaging potentialities relies upon extensively on the evaluation software program. For example, cell hint software program machine studying capabilities dramatically improve goal recognition and reduce gradual turnarounds. Machine studying additionally repeatedly improves course of success.

CellPathfinder offers considerable evaluation capabilities to individualize analysis. It resolves screening bottlenecks and employs AI to simplify superior analyses. It additionally offers detection of uncommon occasions (CTC, for instance) with excessive velocity and excessive accuracy.

We have now additionally labored extensively with co-innovation companions to excellent the atmosphere for particular functions corresponding to colony formation, cytotoxicity, neurite outgrowth, co-culture evaluation, and cell monitoring.

HEK293

Picture of HEK293 taken by the CQ1_50um pinhole. Picture Credit score: Yokogawa Life Science

What future developments in reside cell imaging applied sciences is Yokogawa presently specializing in or planning to discover?

We’re working with co-innovation companions on new functions that may prolong the capabilities of HCA know-how. Plenty of proof-of-concepts are in course of, and, in some instances, researchers have been in a position to report conclusions. For instance, our newest single-cell evaluation answer, the Single Cellome™ System, mannequin SS2000, has been featured in an article revealed within the high-profile journal Analytical Chemistry concerning groundbreaking work performed by researchers on the College of Surrey, England, within the rising area of single-cell lipidomics.

The SS2000 is a reside cell picture machine geared up with Yokogawa’s unique dual-microlens spinning disk imaging know-how, enabling cutting-edge life science analysis.

Lipidomics is the large-scale research of lipids, a big household of molecules that play organic roles starting from sustaining regular operation of the human physique to the event of main illnesses. Lipidomic research are conventionally performed by bulk isolation and evaluation of lipids from many cells.

Whereas bulk lipid analyses can present scientists with a inhabitants common of the lipid profile, they’re unable to discern delicate cell-to-cell variations or reveal the spatial or temporal variations within the lipidome attributable to cell-cell interactions.

Single-cell lipidomics is an rising area the place the lipid composition of single cells is analyzed. These research assist overcome the challenges of bulk lipidomics by providing scientists a way of exploring spatial and temporal variations along with intercellular variability. Understanding these variations is essential to making a extra full image to know illnesses corresponding to most cancers. One problem dealing with researchers is the power to isolate single cells in a manner that maintains the pure lipidome of a cell.

Present strategies of single-cell isolation detach and droop a number of cells directly after which isolate them by way of a slim channel, however this may be notably irritating to cells and should lead to alterations to mobile lipid make-up.

Yokogawa’s SS2000 Single Cellome System makes use of confocal imaging know-how to assist overcome challenges with single-cell lipidomics. It’s a dual-purpose system that permits reside cell imaging and in addition performs absolutely automated single-cell and subcellular sampling with out going by way of the suspension course of, thereby minimizing stress on the cells.

Spinning Disk Confocal Microscopy

Spinning Disk Confocal Microscopy Expertise. Picture Credit score: Yokogawa Life Science

How do partnerships and collaborations play a task in Yokogawa’s technique for growing and enhancing microscopy and imaging options?

Partnerships and collaborations with key stakeholders are integral to Yokogawa’s improvement course of. The corporate makes use of the phrase “co-innovating tomorrow” as a company slogan. It represents the dedication to co-create new worth for fixing clients’ issues and increasing new alternatives whereas cultivating long-term partnerships.

Yokogawa has additionally shifted the route of the corporate’s R&D actions from not solely growing new applied sciences and creating worth by itself to co-creating new worth in cooperation with clients.

The important thing thought is that we should abandon the traditional idea of company-centric worth creation, set up a brand new paradigm of worth co-creation between corporations and clients, and transform enterprise actions to align with them. Moreover, the idea of design considering urges us to not merely derive concepts solely from applied sciences however to go to the sector, observe customers, and create an thought primarily based on empathy.

How have trade conferences and boards influenced Yokogawa’s product improvement methods and enterprise relationships within the scientific group?

The conferences and boards have allowed us to develop our alternatives for co-innovation by way of publicity to new end-users, influencers, and trade leaders. Within the life sciences industries, such occasions have been very energetic. In such a fast-changing trade, it’s vital to work together on a virtually steady foundation.

How does buyer suggestions form the evolution of Yokogawa’s merchandise, notably within the area of confocal microscopy and picture evaluation?

Whereas buyer suggestions is paramount in any trade, we see these fields as quickly evolving and, thus, in want of even nearer work along with clients. There’s virtually a steady course of of latest developments in drug discovery and analysis. We take an method that each one clients are co-innovation companions.

Concerning the Speaker

Kevin Jan presently holds the place of Principal at Yokogawa, the place he leverages his in depth expertise in course of automation and management programs. With a robust background in engineering and a confirmed observe document within the trade, Kevin performs a vital function in driving progressive options and advancing Yokogawa’s technological capabilities. His management and experience contribute considerably to the corporate’s success in delivering high-quality providers to shoppers worldwide.

About Yokogawa Life Science

We have now 30 years of expertise on this life science area and can reply to buyer’s drawback fixing with cutting-edge options.

Our confocal scanner unit CSU collection permits 3D remark of the cells intimately and dynamics of organelles inside cells. Because the CSU collection is able to high-speed taking pictures, it is usually appropriate for observing high-speed life phenomena. As well as, the CSU collection is a multi-point confocal methodology which is extraordinarily mild to cells, finest appropriate for long-term reside cell remark.


LEAVE A REPLY

Please enter your comment!
Please enter your name here

Read More

Recent