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Single Cell Manipulation and Live Cell Imaging
collaboration with CYTOSURGE

Femtoliter Precision Meets Microsecond Accuracy

The FluidFM BOT system was designed and engineered by the Swiss company Cytosurge. As base of the system Cytosurge decided to use Olympus IX83 Imaging platform reknown for its precision and stability.  The Cytosurge FluidFM BOT brings together the best of both worlds, uniting Cytosurge’s innovative microfluidics- and force microscopy-based solution for nano-injection, nano-extraction, cell isolation and nano-printing with Olympus long standing expertise in multi-dimensional live cell imaging.

Femtoliter Precision Meets Microsecond Accuracy

Cell Manipulation Technology

Cell Manipulation Technology
Cytosurge’s patented hollow probe FluidFM technology unites 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.

The World‘s Smallest Syringe

Thanks to the microfluidic channel inside the Cytosurge FluidFM probes, soluble molecules can be dispensed or extracted through a sub-micrometer aperture at the tip (Guillaume-Gentil et al. 2014). At the same time, the sensitive force feedback system allows 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 Syringe

Nano-Extraction

Nano-Extraction
Using the FluidFM nanosyringe, perform gentle biopsies on selected cells. The approach is similar to FluidFM nano-injection but uses negative pressure to extract the cell content. Depending on your interest, target either the nucleus or the cytoplasm of the selected cells. The precise dimensions of the nanosyringe allow you to quantify precisely the volume of the biopsy. The image shows the nano-extraction of 1110 fL from a GFP-Hela cell cytoplasm. The procedure preserves the cellular context and is non-destructive and therefore makes it possible to perform cell biopsies from the same single cell over different time points.

Nano-Injection

With the Cytosurge FluidFM BOT system you can perform nano-injection with a vast variety of materials of your choice selectively into either cytoplasms or nuclei of adherent cells. This is achieved in a cell-context preserving, non-destructive (cell viability ~ 95%), measurable (fl volumes) and fast (inject 100+ cells/hour) manner. Other potential applications of the FluidFM BOT are nano-extraction and cell isolation. Together with FluidFM nano-injection, these applications make the FluidFM BOT an unrivalled tool in many key research areas.

Cell Isolation

Adherent cells can easily be isolated using the Cytosurge FluidFM micropipette. Selectively shower the cell of interest with trypsin or any other dissociation reagent. Then, apply a gentle negative pressure to harvest the cell with the micropipette. You can then move to another well to isolate the cell by approaching the surface and applying a positive pressure through the microfluidic channel of the micropipette.

(image shows the isolation of an adherent CHO cell using the FluidFM BOT)

Cell Isolation

Nano-Printing

Nano-Printing

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 nano-printed to create submicron structures which will act as scaffolds for controlled cell growth.

(Image shows nano-printing of DAPI on DNA coated glass)

Live Cell Imaging Technology

Cell manipulation is only the first step in an experiment and of course analyzing the effects of the manipulation is at least equally important. A variety of scientific questions concerning dose response, gene expression, cell differentiation, intracellular transport, colocalization and more can be addressed with live cell time-lapse imaging techniques.

Olympus not only offers dedicated systems for live-cell imaging, confocal imaging and high content screening but is also able to equip the FluidFM BOT 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 needs to be accurately compensated for. Olympus’ laser-based TruFocus system ensures that samples are always in focus, therefore producing the sharpest images possible. The near-infrared laser continuously checks and, if required, readjusts the focal position, making sure cells are always in focus even in long term time lapse experiments.

Microsecond Accuracy for Increased Cell Viability

The Olympus Real-Time Controller (RTC) features an independent CPU board that guarantees fast, parallel experiment execution without delays even in complex experimental set-ups. This leads not only to extremely high timing accuracy and precision of smaller than 2us which is essential for high speed imaging.

Precise device control, excellent timing precision and high accuracy are prerequisites which reduce sample bleaching, maximize cell viability and guarantee data reproducibility.

Learn more about live-cell imaging

Related Systems

FV3000

FV3000

Confocal Laser Scanning Microscope

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IXplore

IXplore

Inverted Imaging Systems

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scanR

scanR

High Content Screeinng

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References

Cell Manipulation:

Nano-injection:

Nano-extraction:

Cell Isolation:

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