Examine Cell Mechanics With Fast Single Cell Force Spectroscopy

What promotes cell adhesion, what reduces it? And by how much? Working with cell cultures, adhesion or the lack thereof has great impact on culture viability, growth speed, and even differentiation. Using unique microfabricated and force-sensitive pipettes, the adhesion of cells can now be quantified much faster and studied on the single cell level with FluidFM technology.

A single cell is detached from fully adherent and confluent culture. The measured forces both depend on the substrate and the bonds to the neighboring cells. Courtesy of A. Sancho and J. Groll; Functional Materials for Medicine and Dentistry, University Hospital of Würzburg.

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Single cell force spectroscopy provides excellent mechanical data on a single cell level. To date, its main disadvantage has been its low throughput and complicated handling. Now, FluidFM-based single cell force spectroscopy provides 10 times higher throughput, as well as greatly simplified manipulation of the cells under investigation.

Pick a cell with negative pressure, measure, release it again with a positive pressure pulse or by a short cleaning procedure.

Choose a cell with negative pressure and then measure. Re-release it with a positive pressure pulse or by a short cleaning procedure. Image Credit: Cytosurge AG.

FluidFM is able to process and provide data on 20 to 200 cells a day, ten times faster than conventional methods. Additionally, its force range is ten times higher than standard methods, with pN resolution.

FluidFM for cell mechanobiology and biophysics

The physical study of single cells offers insights into biological phenomenon such as differentiation, growth and proliferation in both biophysics and mechanobiology. It is essential to gather deeper insight and gain more advanced understanding of mechanical cell properties and their interactions with the surrounding environment in areas ranging from cancer research to stem cells and organoids. There is a clinical need in the case of implant materials to gain intimate knowledge of and firm control over how cells adhere to the material in question.

With the heterogeneity of cells, meaning every cell is different from its neighbour, one goal is to gain understanding of the effects of these differences on a single cell level. Single cell force spectroscopy has been developed as an insightful method to investigate these questions using atomic force microscopy (AFM).

However, the process of gluing a cell to an AFM cantilever is considerably time-consuming, which limits the daily throughput to just a few cells. But, typically, because of cell-cell heterogeneity, numerous cells are needed to meaningfully assess any experimental condition.

In this instance, FluidFM provides significant improvements by reversibly immobilizing a cell to a FluidFM probe by suction, and subsequent release with pressure. This enables users to increase the throughput by a factor of 10 to 100.

A cell is detached from adherent culture with a FluidFM micropipette.

A cell is detached from adherent culture with a FluidFM micropipette. Image Credit: Cytosurge AG.

FluidFM has provided us the chance to detach mammalian cells that were very strongly adhered to the substrate, in a systematic way and without any chemical modification of the cantilever; thus, allowing us to study cell behaviour in their natural state and environment.”

A. Sancho, University Hospital of Würzburg and University of the Basque Country.

FluidFM Features

Reduced processing times with FluidFM

Shorter preparation times working alongside reusable measurement probes means FluidFM is the perfect tool for all types of single cell mechanical studies. Due to the unique properties of FluidFM technology, mechanical data on solid cells can be collected in greatly reduced timeframes. Users are able to gain access to unmatched measurement ranges, which increases experimental flexibility.

Measure suspended or fully adherent cell

FluidFM micropipettes allow users to attract a suspended cell from solution or to pick it up from fully adherent culture, and as a result, FluidFM can work successfully with cell adhesion forces of over 1,000 nN.

From pN to µN

FluidFM allows users to measure forces from tens of pN up to µN by taking advantage of the FluidFM probes, that are made in multiple grades of stiffness and opening diameters. A wide range of cells and mechanics can be investigated with spring constants from 0.3 N/m to 4 N/m and openings from 300 nm to 8 µm.

Fast cleaning between cell measurements

With many mammalian cells adhering quickly to any kind of substrate, including the FluidFM probe, users will often clean the probe after measuring each cell. With FluidFM, this procedure takes no longer than two minutes before the next cell can be picked for measurement. This fast cleaning process can be skipped for non-adherent cells, making the overall throughput even higher.

Illustration of the detachment of a cell from a substrate by FluidFM. The measured force-distance curve is able to provide insights on adhesion strength, energy, and distance, as well as the involved bio-chemical bonds.

Illustration of the detachment of a cell from a substrate by FluidFM. The measured force-distance curve is able to provide insights on adhesion strength, energy, and distance, as well as the involved bio-chemical bonds. Image Credit: Cytosurge AG.

FluidFM ADD-ONs for AFM

Users are able to carry out FluidFM single cell force spectroscopy experiments, and many more applications, by adding a FluidFM ADD-ON to their AFM. It is available for a range of AFMs.

FluidFM Probes

The unique, patented FluidFM probes, can be used for adhesion experiments and a range of other manipulation tasks. They come in a wide range of types and specifications, tailored for each application.


Force spectroscopy can be carried out with the FluidFM ADD-ON for AFM, how exactly - and much more - can be found here.

FluidFM in publication

FluidFM has been featured in a number of publications in order to quantify cell mechanical properties. Although there are many more in the publications section, below are three publication highlights.

Optimizing drugs

Millions of people are diagnosed with leukemia every year worldwide. Although treatment drugs are used, cancer often becomes resistant against them. Researchers from the University Hospital Würzburg and the University of Würzburg discovered a potential approach that may overcome resistance to the recently approved Midostaurin drug, with the possibility of even increasing the drug activity. This was possible with the help of FluidFM cell measurements.

Optimizing stents

Stents save countless lives every year and help millions of people to overcome arterial blockages. Once implanted, the stent should integrate well and stop blood clots from forming and blocking arteries. Research groups from ETH Zurich investigated stent design optimization in this publication using FluidFM to measure cell adhesion to its surface.

Calibrating high-throughput devices

Despite offering crucial insights into a wide range of fields, single cell force spectroscopy is held back by its low throughput. MTA Budapest significantly excels the acquisition of single cell adhesion data by pairing with FluidFM adhesion measurements in order to calibrate an optical sensor array. As a result, researchers are able mechanically monitor over 1,000 adherent cells in parallel.

About Cytosurge AG

Our FluidFM solutions bring significant benefits to a wide range of applications in life sciences, biophysics and mechanobiology. Benefit from quantitative volume measurements of injected compounds during drug development, improved CRISPR gene editing by direct delivery into the nucleus, 2.5D nano-printing down to sub-micron levels or single cell adhesion and colloidal probe measurements.

Cytosurge AG develops, manufactures and distributes state-of-the-art nanotechnology solutions and systems based on its patented FluidFM® technology. At the heart of the technology are the patented hollow FluidFM probes which have apertures down to 300 nm enabling the handling of femtoliter volumes.

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Last updated: Mar 31, 2020 at 9:49 AM


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