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.
A1R HD25: the latest in resonant scanning technology allows new live-cell imaging approaches
Capturing the dynamics of living systems requires high acquisition rates. Large samples, such as whole model organisms, additionally require a large field of view. The Nikon A1R HD25 confocal system provides both, combining Nikon’s improved HD high speed resonant scanner with an unprecedented 25 mm field of view. The performance of this system is evaluated in zebrafish embryos.
Antitumor Lymphocyte Kinetic Cytotoxicity Assay (Collaboration with CCRM)
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.
Automated, Non-Invasive Culture, and Evaluation System for iPS Cells under Neural Differentiation Process
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
Establishment of an Algorithm for Automated Detection of iPS/non-iPS Cells Under a Culture Condition by Non-Invasive Image Analysis
Generation of a Growth Curve for iPS Cells in a Feeder-Free Culture by Non-Invasive Image Analysis
Increasing Data Collection and Fidelity by Maximizing Confocal Field of View
For years, the field of view (FOV) of confocal systems has been limited by the FOV of the microscope they are attached to. With the release of the Nikon Ti2 inverted microscope, the world’s first 25-mm FOV became available. Now, Nikon has taken advantage of this improvement by building the largest FOV point scanner in the world, the A1 HD25. This Application Note focuses on the impact of this technology on simple, everyday experiments.
N-SIM for Quantitative Ultra-Structural Analyses of the Nuclear Lamina
Super-resolution Structured Illumination Microscopy (SIM), available from Nikon via the N-SIM and N-SIM E systems, allows for the observation of details inaccessible to traditional microscopes, such as confocal and widefield. In this application note we see how the N-SIM system enables quantitative multi-color evaluation of the distribution of different nuclear lamin proteins and the structures they form.
N-STORM with DNA-PAINT for Reliable Multicolor & 3D Single Molecule Localization Imaging
Nikon’s Large-Format Multiphoton System for Intravital Imaging
Nikon’s A1R MP multiphoton confocal microscope system provides a choice platform for deep tissue imaging. Researchers at the Allen Institute for Brain Science have applied the A1R MP towards fast dual-color intravital imaging of the mouse brain. The open architecture instrument design allows for the addition of custom stages for large samples, external photostimulation devices, and more – perfect for constantly evolving needs.
Outline of Stem Cell Applications
Quantitative Analysis Tools and Correlative Imaging Applications for N-STORM
Stochastic Optical Reconstruction Microscopy (STORM) has had a significant impact on our understanding of nano-scale biological processes. In this application note, we see how Nikon’s N-STORM system, combined with quantitative analysis tools, enables single molecule level studies of cannabinoid signaling in the brain. Correlative 3D-STORM, confocal microscopy, and patch-clamp electrophysiology is also explored.
Quantitative Cluster Analysis Applications for N-STORM
Nikon’s N-STORM super-resolution microscope is applied to mapping the distribution of nucleosomes on chromatin fibers, shining a light on the mechanisms of chromatin folding, gene expression, and pluripotency. Multicolor STORM is used to explore how nucleosome distribution differs between differentiated and pluripotent stem cells and correlates with the distribution of other important factors in replication, such as RNA Polymerase II.
Reflectance imaging for visualization of unlabeled structures using Nikon A1 and N-SIM
Reflectance imaging allows the user to form an intensity image from light backscattered by the sample. Highly reflective markers, including a variety of nanoparticles, allows for imaging with very high signal-to-noise and virtually free of photobleaching, ideal for both confocal and structured illumination microscopies.
Screening Assay for PSC Responses to Substrates
Stem Cell Research with Nikon
Structured Illumination Microscopy (SIM) Imaging Comparison with Confocal
The super-resolution microscopy technique structured illumination microscopy (SIM) imaging of dendritic spines along the dendrite has not been previously performed in fixed tissues, mainly due to deterioration of the stripe pattern of the excitation laser induced by light scattering and optical aberrations.
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.