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CL-Quant Add-on Modules
In place of visual judgment, the application enables objectively measurement, quantification and record of the adherent cell occupancy ratio (confluency) that is important for determination of the timing of passages and assays.
Human pluripotent stem cell (hPSC) colony coverage area ratio
The application enables quantification and record of the ratio of coverage area (confluency) of hPSC(human iPS/ES cell) that form characteristic colonies.
Human pluripotent stem cell (hPSC) colony number
The number of hPSC(human iPS/ES cell) colonies is countable by automatic calculation without complicated work.
Human pluripotent stem cell colony (hPSC) application package
The following four items on hPSC(human iPS/ES cell) are measured.
- Colony number
- Individual colony area
- Average colony area
- Total colony area
Neurite length (phase-contrast image)
The application can be measured the total lengths of neurites in the field automatically from phase-contrast images without staining.
Neuronal cell and/or cluster number (phase-contrast image)
The application can be measured the neuronal cell and/or cluster numbers in the field automatically from phase-contrast images without staining.
Neurite length (fluorescence image)
The application can be measured the total lengths of neurites in the field automatically from fluorescence images of cells in which neurites or whole neurons exhibit fluorescence.
Neuronal cell and/or cluster number (fluorescence image)
The application can be measured the neuronal cell and/or cluster numbers in the field automatically from fluorescence images of cells in which neurites or whole neurons exhibit fluorescence.
Survival Analysis of Human In Vitro-derived Neurons Using New Live Cell Extended Time-lapse Imaging Technology
In this application note we demonstrate the Nikon BioStation CT for long-term live-cell imaging of stem cell-derived motor neurons – tracking key attributes such as cell body size and neurite length before, during, and after treatment with different stressors. This approach provides a standardized method for drug discovery studies in neurodegenerative disease models.
Development of an Integrated Bioprocess for Production of NK-92 Cells for Immunotherapy
Anti-cancer NK-92 lymphocytes are promising for application in immunotherapies. This application note explores a comprehensive workflow for producing clinical-grade NK-92 cells, using Nikon’s BioStation CT to monitor NK-92 phenotype and potency.
Embryonic Stem Cell Responses to Commercially Available Substrates for Stem Cell Maintenance
In collaboration with the Centre for Commercialization of Regenerative Medicine (CCRM) we compare the performance of 8 common substrates for stem cell growth and maintenance by developing an assay utilizing the Nikon BioStation CT high content screening microscope and incubation system. Fluorescence imaging of Oct4 is used as a marker for pluripotency.
Establishment of an Algorithm for Automated Detection of iPS/non-iPS Cells Under a Culture Condition by Non-Invasive Image Analysis
In this application note we detail an automated high throughput method for observing and classifying stem cell colonies with a standardized algorithm and using the Nikon BioStation CT. Automated image analysis discriminates stem cell colonies from non-stem cell colonies using phase contrast imaging and subsequent analysis of distinguishing morphological features.
Automated, Non-Invasive Culture, and Evaluation System for iPS Cells under Neural Differentiation Process
This application note details an integrated workflow for the culture, observation, evaluation, and differentiation of induced pluripotent stem (iPS) cells into neurons, introducing image processing methods for detecting neural rosettes and neurite outgrowth – two important morphological checkpoints in neural differentiation. Automated imaging for this assay is performed using a Nikon BioStation CT.
Generation of a Growth Curve for iPS Cells in a Feeder-Free Culture by Non-Invasive Image Analysis
Long-term time-lapse observation of feeder layer-free stem cell cultures, along with automated detection of stem cell colonies, is demonstrated here using a Nikon BioStation CT system. Growth curves for both differentiated and un-differentiated cells were generated for a time period of about 1 week.
A Non-Invasive Method for Counting Human Pluripotent Stem Cell Numbers by Live Cell Imaging
Presented here is a method for counting live pluripotent stem cells using the fluorescent nuclear stain SYTO24 and phase contrast imaging on the Nikon BioStation CT high content screening microscope. The results show that colony coverage area deduced from phase contrast images can be used as an accurate estimator of stem cell count alone, correlating well with haemocytometer-based measurements.
Automated Haematopoietic Colony Forming Cell (CFC) Assay
Nikon’s BioStation CT high content live cell screening system is applied in an assay for automated enumeration and identification of haematopoietic colony-forming cells. This assay allows for the identification of multiple colony types, helping distinguish between lineages.
Antitumor Lymphocyte Kinetic Cytotoxicity Assay (Collaboration with CCRM)
In this application note we explore the use of the fully robotic Nikon BioStation CT high content screening microscope platform towards quantifying the death rate of a model human leukemia cell line upon co-incubation with anticancer NK-92 lymphocytes. This assay integrates automated imaging of Calcein-AM fluorescence to perform live/dead cell analysis.
Screening Assay for PSC Responses to Substrates
An automated imaging-based screening assay for assessing stem cell pluripotency is described. Researchers from Nikon and the Centre for Commercialization of Regenerative Medicine use a Nikon BioStation CT with CL-Quant software to perform stem cell pluripotency maintenance assays in experiments testing feeder cell-free culture vessel substrates. Such commercially available substrates provide a number of process standardization benefits.
Outline of Stem Cell Applications
The Nikon BioStation CT cell culture observation system and CL-Quant software provide a powerful platform for stem cell research applications, including automated induced pluripotent stem (iPS) cell colony identification, proliferation analysis, and neural differentiation analysis. Real experimental data is provided for each of these use cases.