The intuitive and easy to learn SYNENTEC YT-software accompanies the CELLAVISTA 4 and NyONE imagers. It meets the highest standards of automated imaging, image acquisition, device controlling, and image and data analyses.
The YT-software is a complete tool: starting from experiment set-up, automatic high throughput image acquisition, precise image analysis, storage, and processing of data. Once the high resolution images are captured, they can be viewed and (re-)analyzed in the future.
Data generation in CELLAVISTA4 and NYONE is conducted using one of SYNENTEC’s 65 proprietary image processing algorithms. The high-quality images are analyzed for specific properties, e.g., cells detection or cell features, and the results are visualized using overlays in YT-software.
Applications:
SYNENTEC’s automated imagers are indispensable in cell line development, basic and translational oncology and hematooncology, preclinical drug development, cell and molecular biology, virology, immunology, stem cell research, biomarker research, neurosciences, and cell and gene therapy.
CELLAVISTA 4 and NyONE are versatile automated imagers that currently support over 60 applications, including:
Featured Applications:
Single Cell Cloning - Proof of Monoclonality
Single cell cloning (SCC) represents a crucial step in cell line development. The aim of SCC is to identify wells where a single cell was seeded and to monitor its growth rate. To prove monoclonality for regulatory approval, it is essential to have a robust and reliable imaging system. CELLAVISTA 4 and NYONE automated cell imagers are well established in the field of cell line development. They combine high throughput and accuracy with high image resolution (below 1 micron per pixel) to determine the clonality of a cell line.
Single Cell Cloning (SCC) Using Brightfield Imaging
Benefits:
- Rapid cell cloning.
- No area in the well is missed.
- Day-by-day automatable measurements.
- All colonies are recognized by the YT- software.
- Tracking back to the first day to demonstrate monoclonality.
- Works with both adherent and suspension cells.
- This provides a proof that your colony in fact grows from a single cell, or grows from a cell double.
- Automatic generation of a clone gallery.
Fluorescence Activated Single Cell Cloning (FASCC)
Ghost cells represent one of the main challenges in cell line development. Such undetected cells usually lie on the well edge. They are sometimes missed because most imaging systems for CLD are equipped with low resolution optics (2 microns per pixel), and only have a white light (brightfield) source. This lack of resolution makes it difficult to distinguish cells from debris, or well edge artefacts. Because of this lack of precision, a correct statement about monoclonality is difficult.
Adding fluorescence enables the detection of these cells in a reliable way, even at the well edges!
SYNENTEC has developed a unique method called FASCC to overcome all disadvantages of classic white light single cell cloning. FASCC provides a fully automated screening of your microplates by using the advantage of short term, non-toxic, and animal-free cell staining (e.g., CellTracker Green, Calcein AM). This unique tool presents a fast proof of monoclonality without guesses or doubts!
FASCC Workflow
On the day of cell seeding, two rapid scans are performed
Pre-scan: a fast, low resolution (4x lens) high speed scan to detect the fluorescence signal of e.g. Calcein-AM or Cell Tracker® stained cells. The embedded image analysis tool excludes all wells seeded with more than just one cell (or any number of cell-count the user specifies).
Nanoview: all wells selected after the pre-scan are imaged at high resolution (10x lens) simultaneously in two channels (brightfield and fluorescence channels); with one image per well, and the single cells being automatically in the center of images. This process facilitates the elimination of cell doublets.
Confluence: brightfield images of the entire wells with a documented single cell per well are generated to automatically detect colonies and to monitor the colony growth.
HIGH-THROUGHPUT TRYPAN BLUE DYE EXCLUSION ASSAY FOR MEASURING CELL COUNTS AND VIABILITY
- A widely used technique to determine cell numbers and viability
- Very fast cell counting: 96 samples in 3 minutes!!!
- Small sample volumes, as low as 40 µL
- Brightfield imaging
- Using standard 96-well or 96-well half-area microplates
- A very low cost per sample
- Accurate and reproducible data
PRECLINICAL DRUG TESTING
The development of new drugs and biopharmaceuticals requires precisely conducted experiments. Traditionally, the steps to reach this goal involved time-consuming and labor-intensive laboratory testing. The automation of this process increases the throughput and quality of data, minimizes technician’s errors, and significantly increases the productivity of the lab.
SYNENTEC’s CELLAVISTA 4 and NyONE automated imagers support the following applications in preclinical drug testing:
- High-throughput trypan blue dye exclusion assay
- Cell confluence to measure adherent cells
- Live/dead cytotoxicity assays
- Apoptosis assays (Annexin V, JC-1, caspases)
- Detection and quantification of circulating tumor cells (CTCs)
- Antibody and ADC internalization
- Immunocytochemistry (ICC)
- Cell cycle assays
- DNA damage assay
- CD marker analyses
- 3D culture assays
AUTOMATED WOUND HEALING ASSAY
Cell migration assays are very important tools to study the invasion processes and metastasis of cancer cells. They are also used in arthritis and osteoporosis research. The simplest approach to monitoring a cell migration is the so-called “scratch” assay. A pipette tip is used to scratch or “wound” a confluent cell monolayer. Then a microscope is used to observe cells filling in or “repairing” the wound.
SYNENTEC, in collaboration with the team led by Prof. Susanne Sebens from the Institute for Experimental Cancer Research, CAU + UKSH Kiel, Germany, developed the fastest and most flexible automated wound healing application.
VIROLOGICAL RESEARCH AND VACCINE DEVELOPMENT
SYNENTEC’s automated imagers are frequently used in virology and human and veterinary vaccine development (influenza, Noro, SARS-CoV-2, malaria, HIV, etc.). CELLAVISTA 4 and NyONE cell imagers greatly accelerate your work and support the following assays:
- Viral titer quantification & infectivity determination
- Label-free plaque assay
- Endpoint dilution assays (TCID50)
- Plaque reduction neutralization test (PRNT) (Virus neutralization assay)
- Focus forming assay (FFA)
- Immunofluorescence foci assays (IFA)
- Measurement of host cell viability & viral cytopathic effects (Cell confluence, Live/Dead, Apoptosis)
- Viral Pathogenesis (morphological change in infected host cells)
- In vitro immune responses to vaccines
- Vaccine vector testing in transduction efficiency
- Immunologic in vitro studies
- Reverse vaccinology (ELISpot and FluoroSpot)
3D cell culture assays
The complexity of tumorogenesis is not reflected in conventional two-dimensional (2D) monolayer cell culture models. Therefore, three-dimensional (3D) cell culture models have gained interest in recent years. In these 3D models, cells do not grow as one layer of cells but form a complex three-dimensional structure. This structure provides a more accurate representation of tumor physiology as occurring in cancer diseases.
Quantification Of 3D Spheroids Using Brightfield Channel
SYNENTEC’s Spheroid Count operator analyzes spheroids and quantifies the size and number of spheroids per well. High-throughput non-invasive brightfield imaging is implemented.
High-Content Spheroid Analysis – Multiparametric Evaluation
Spheroids are produced in Kugelmeiers’ Sphericalplate 5D 24-well plate. Each well contains 750 round-bottom microwells that produce uniform spheroids. Spheroids are imaged by CELLAVISTA 4/ SCIENTIFIC and NyONE/ SCIENTIFIC imagers that use harmonic drive without turbulence. High-content and high-throughput analyses of these 3D spheroids are performed using a combination of fluorescence dyes.
More information about imaging 3D cell culture assays is available on Synentec’s website.