Single Cell Manipulation and Live Cell Imaging
collaboration with Cytosurge

Olympus is proud to partner with Cytosurge to become a complete system provider to the scientific community’s growing need for next-generation single-cell and CRISPR genetic manipulation solutions.

The demand continues to rise as researchers see their potential to study diseases, find medical treatments, and benefit human health. Cytosurge provides a powerful genetic manipulation solution with the FluidFM® OMNIUM (previously called FluidFM BOT BIO Series), a highly automated system built on the Olympus IX83 inverted microscope with Olympus' renowned optics. In addition to CRISPR genome editing, the system enables unprecedented research across a wide range of applications. In neuroscience, for instance, researchers can generate patterned neuronal networks, and in single-cell omics, they can perform repetitive extractions of cytosol from the same cell.

FluidFM OMNIUM System—Femtoliter Precision Meets Nanometer Accuracy

The FluidFM OMNIUM system was designed and engineered by the Swiss company Cytosurge. For the system base, Cytosurge uses the Olympus IX83 inverted microscope, an imaging platform known for its precision and stability. The FluidFM OMNIUM system unites Cytosurge’s innovative microfluidics- and force-microscopy-based solution for single cell research with Olympus’ long-standing expertise in multidimensional live-cell imaging.

Femtoliter Precision Meets Microsecond Accuracy

Core of the FluidFM OMNIUM system. Courtesy of Cytosurge AG.

Direct intra-nuclear injection with FluidFM technology. Courtesy of Cytosurge AG.

FluidFM Technology—For Single-Cell Manipulation

Cytosurge’s patented hollow probe FluidFM technology combines the optimal features of microfluidics and force microscopy by introducing closed microscopic channels into force sensitive probes. These probes have apertures down to 300 nm, enabling them to simultaneously sense interaction forces down to pN and to dispense or aspirate femtoliter volumes.

FluidFM Modes—For a Variety of Applications

The different tip shapes, aperture sizes, and mechanical specifications of FluidFM probes support three basic modes—pick and place, injection and sampling, and localized dispensing—enabling innovative new applications in neuroscience, virology, mechanobiology or CRISPR cell line development.

Pick and Place

Grab and relocate single cells and
other micro-sized objects.

Injection and Sampling

Inject into and extract from single cells with the possibility to quantify the injected or extracted volume.

Localized Dispensing

Dispense a tiny amount of liquid,
in air or in immersion.

Neuroscience

FluidFM technology is a novel and innovative toolbox that opens up an array of possibilities for single-neuron manipulation. By uniting the optimal features of microfluidics and force microscopy and using different force-controlled probes, FluidFM technology provides a wide range of innovative methods, from controlled patterned growth to single cell manipulation, stimulation, and analysis, in a gentle manner suited for sensitive cells such as neurons.

  • Pattern
    Define where axons will grow.
  • Pick and Place
    Build microbrains by creating neuronal networks.
  • Stimulate
    Apply neurotransmitters anywhere on the neuron.
  • Inject
    Deliver CRISPR complexes directly into the nucleus.
  • Observe
    Track your manipulated neuron over time.
  • Analyze
    Extract cellular content while keeping the neuron intact and alive.

Stimulate, inject into, and observe single neurons with FluidFM technology. Courtesy of Cytosurge AG.

Direct intra-nuclear injection of CRISPR complexes with FluidFM. Courtesy of Cytosurge AG.

CRISPR Cell Line Development

Overcome CRISPR delivery limitations with Cytosurge’s FluidFM technology. FluidFM offers a unique in vitro solution to improve the efficiency and applicability of CRISPR across a variety of cell types and enables cell-line development in less than 3 weeks.

  • Gentle
    Ultragentle transfection method based on nanoinjection.
  • High efficiency
    High CRISPR HDR efficiency due to direct intranuclear delivery.
  • For sensitive cells
    Especially suitable for hard-to-transfect and rare cell types.
  • Multiplexing
    Easily introduce multiple gene edits in one go.
  • Fast cell line development
    Generate stable monoclonal cell lines in less than 3 weeks.

Virology

Go beyond bulk experiments—study viral entry and replication on a single virion-single cell level. With the gentle, microchanneled, and force feedback-controlled FluidFM probes, single virions can be deposited onto selected cells, enabling unprecedented control to study viral infections in vitro. FluidFM technology can therefore help to illuminate fundamental questions on virulence, virus replication, or host immune response, boosting the development of novel antiviral drugs and vaccines.

Working principle of viral manipulation with FluidFM technology. Courtesy of Cytosurge AG.

  • Precisely deposit a single virion
    Single virions can be placed at an exact location on your cell of choice.
  • Inject a single virion
    Directly inject a single virion into the cytoplasm or the nucleus of a specific cell.
  • Measure biophysical changes
    Measure mass changes, variation in stiffness, and changes in adhesion force.
  • Isolate, extract, and analyze
    Isolate infected cells—or take single cell biopsies—for further expansion or analysis.
  • Observe and monitor
    Continuously monitor the cell and observe the surrounding culture via integrated microscope and tracking software.

Mechanobiology

While traditionally a cell needed to be glued to an AFM cantilever, Cytosurge’s FluidFM technology reversibly immobilizes a cell to a FluidFM probe via suction, and subsequently releases it with a pressure pulse or brief washing. This gentle exchange of the cell enables the cantilever to be reused for long periods, saves time and costs, and results in a high throughput, increasing your productivity by more than 10 times* compared with conventional force spectroscopy methods. Measure mechanical data of up to 200 cells a day thanks to reversible cell immobilization with FluidFM technology.

  • Simple and reversible immobilization
    Reversible attachment of object to cantilever by suction.
  • Huge force range
    Measure forces from pN up to µN.
  • Many cell types and colloids
    For mammalian cells, microbes, and colloids.

Working principle of single cell force spectroscopy with FluidFM techology. Courtesy of Cytosurge AG.

* Compared with the 1–3 cells per day measurement typical of conventional force spectroscopy.

Nanoprinting

FluidFM technology with its microfluidic probes offers unprecedented versatility and control to print patterns and explore nanostructure creation. Whether you are working with protein arrays for sensing applications, chemical gradients for cell migration studies, want to understand chemical processes in femtoliter droplets, or are interested to develop the next generation of nanofabrication processes—FluidFM technology opens new doors for your research.

  • From femtoliter to nanoliter
    Print femtoliter droplets or deposit up to nanoliter volumes.
  • From nm to cm
    Print features as small as 300 nm and create patterns up to cm in scale.
  • 1 to 10 000 cP viscosity
    Users have printed with inks from 1 to 10 000 cP, from water to honey.
  • In air and in liquid
    FluidFM technology works both immersed and in ambient conditions.
  • Wide range of inks
    Water, buffers, oils, acids, solvents, nanoparticles, and more.

Single Cell Omics

FluidFM technology provides a fascinating solution for nondestructive and live single-cell omics. Thanks to its gentle cell manipulation technology, FluidFM technology enables the extraction of subpicoliter volumes from the cytoplasm or nucleus of a single cell and the isolation of the extracted contents for further analysis. This cytoplasmic biopsy is so gentle that the cells survive, and sequential biopsies can be performed on the very same cell. By avoiding the destructive cell lysis, trajectory instead of end-point analysis becomes possible, whether in transcriptomics, metabolomics, proteomics, or any other omics studies.

  • Nondestructive extraction
    Gently extract from cytoplasm or nucleus while keeping the cell alive and fully viable.
  • Save the physiological context
    During extraction, keep the targeted cell in its context next to its neighboring cells and conserve established cell-cell interactions.
  • Time-course biopsy
    Repeat the gentle extraction several times on the same cell, e.g., before and after stimulation by a specific drug.
  • Pick and place
    If full cell content is required, isolate the cell directly from an adherent or suspension culture without affecting neighboring cells.

Extraction of cellular content with FluidFM. Courtesy of Cytosurge AG

Live-Cell Imaging Technology

Cell manipulation is only the first step in an experiment. Analyzing the effects of the manipulation is just as important. Live-cell time-lapse imaging techniques can address a variety of scientific questions concerning dose response, gene expression, cell differentiation, intracellular transport, colocalization, and more.

Olympus not only offers dedicated systems for live-cell imaging, confocal imaging, and high-content screening but also equips the FluidFM OMNIUM system with high-end live-cell imaging capabilities.

Maintain Long-Term Focus

In long-term time-lapse imaging, maintaining the focal position is pivotal, and thermal drift introduced by ambient temperature change must be accurately compensated. Olympus’ laser-based TruFocus™ Z-drift compensation system helps ensure that samples are always in focus to produce the sharpest images possible. The near-infrared laser continuously checks and, if required, readjusts the focal position, making sure that cells remain in focus during long-term time-lapse experiments.

Microsecond Accuracy for Increased Cell Viability

The Olympus real-time controller (RTC) features an independent CPU board that delivers fast, parallel experiment execution without delays, even in complex experimental setups. This leads to high timing accuracy and precision of smaller than 2 µs, which is essential for high-speed imaging.

Precise device control, excellent timing precision, and high accuracy are needed to reduce sample bleaching, maximize cell viability, and provide data reproducibility—and the RTC delivers all three.


For more information on the different applications, visit www.cytosurge.com/applications

References

Technology

Neuroscience

CRISPR cell line development

Injection and Isolation publications:

Virology

Mechanobiology

Nanoprinting

Single cell omics

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