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® 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’s core functionalities—intracellular- and nuclear-injection and extraction, pick-and-place at the microscale, and precise dispensing of liquids—enable a broad range of applications, including single-cell drug assays, monoclonal cell line development, and nanoprinting of custom patterns.

Intracellular Injection and Extraction

-Single-cell CRISPR delivery

-Single-cell drug assays

-Single-cell biopsy

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Pick and Place at the Microscale

-Cell isolation

-Direct cell adhesion assays

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Precisely Dispense Liquids
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-Spotting

-Line printing

-Nanoprinting of custom patterns

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Live-Cell Imaging
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-Live-cell time-lapse imaging capabilities

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Femtoliter Precision Meets Microsecond Accuracy

The FluidFM BOT BIO Series 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 Cytosurge FluidFM BOT BIO Series 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

Cell Manipulation Technology

Cell Manipulation Technology Cytosurge’s patented hollow probe FluidFM technology combines the best features of microfluidics and force microscopy by introducing closed microscopic channels into force sensitive probes. This unique combination enables the handling of liquid volumes at the femtoliter scale, as well as force-controlled manipulations of microscopic objects. FluidFM technology elevates the application scope of traditional glass micropipettes to new levels.

Submicrometer Syringe

Thanks to the microfluidic channel inside the Cytosurge FluidFM probes, soluble molecules are dispensed or extracted through a submicrometer aperture at the tip (Guillaume-Gentil et al. 2014). At the same time, the sensitive force feedback system enables the precise and gentle manipulation of cells in a controlled fashion. It also provides a reliable distinction between gentle contact with cell membranes and perforation.

The World‘s Smallest SyringeSoluble molecules are dispensed or extracted through a submicrometer aperture at the tip
(Guillaume-Gentil et al. 2014)

Intracellular Injection and Extraction

Injection

With the Cytosurge FluidFM BOT BIO Series system, you can perform nano-injection with a wide variety of materials selectively into either cytoplasms or nuclei of selected adherent cells. This is achieved in a cell-context preserving, nondestructive (cell viability ~ 95%), measurable (fl volumes), and fast (inject 100+ cells/hour) manner.

Applications

Single-Cell CRISPR Delivery

A solution to CRISPR's delivery challenges: Place your complexes directly where they are required, in the nucleus.

Get a multiple knock-out clone in two weeks, with one instrument.

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Single-Cell Drug Assays

Deliver compounds directly into a cell and quantify the injected volume with femtoliter precision. A suitable method for any compound and easy to use.

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Extraction

Nano-Extraction

Nano-extraction of 1110 fl from a GFP-Hela cell cytoplasm.

Using the FluidFM nanosyringe, perform gentle biopsies on selected cells. This approach is similar to FluidFM nano-injection but uses negative pressure to extract the cell content. Depending on your requirements, you can target either the nucleus or the cytoplasm of the selected cells. The precise dimensions of the nanosyringe enable you to precisely quantify the volume of the biopsy.


Pick-and-Place at the Microscale

Reversibly immobilize cells and other objects by gently applying suction force. A gentle negative pressure through the microfluidic channel of a FluidFM probe enables you to pick up and hold a cell for cell isolation or adhesion measurements. Simply select a specific cell directly from a culture with a mouse click. For suspended and adherent cells.

Cell Isolation

The isolation of an adherent CHO cell using the FluidFM BOT BIO Series.

Applications

Cell Isolation

Choose the cell you want – place it where you want.

  • Create in-vitro neuron networks
  • Single-cell co-culture for intra-cellular communication
  • Rare cells sorting
  • Gateway to single @cell omics
  • Generate monoclonal cell cultures
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Single Cell Force Spectroscopy

Easily measure forces.

  • From confluent layers
  • Single cells
  • 100s of cells a day, compared to tens reached by conventional means
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Precisely Dispense Liquids

Nano-PrintingNanoprinting of DAPI on DNA-coated glass

Print spots, high-density arrays, or even complex patterns with nanometer precision, useful, for example, in biosensing to create protein or DNA arrays. Also Poly-L-Lysine can be nanoprinted to create submicron structures that will act as scaffolds for controlled cell growth.

Applications

Spotting

Precisely deposit a thousand spots of any solution. Ideal for developing and testing protein-based assays.

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Line Printing

Print lines with widths from tens of micrometers down to hundreds of nanometers.

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Nanoprinting of Custom Patterns

Print custom patterns from a file.

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Virus Deposition

Place virions one-by-one with pin-point accuracy onto single cells.

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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 BOT BIO Series systems with high-end live-cell imaging capabilities.

Live Cell Imaging Technology

Never Lose 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™ drift compensation 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 cells are always 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 extremely 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.

Learn more about live-cell imaging

Related Systems

FV3000

FV3000

Confocal Laser Scanning Microscope

Learn more

IXplore

IXplore

Inverted Imaging Systems

Learn more

scanR

scanR

High-Content Screeinng

Learn more

References

Cell manipulation:

Injection:

Extraction:

Pick and Place at the Microscale:

Precisely dispense liquids::

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