10x Faster Single Cell Force Spectroscopy with Your AFM

Get Sound Statistics Within A Day

Single cell force spectroscopy provides quality mechanical data on a single cell level, however its low throughput and complex handling is a significant drawback. FluidFM-based single cell force spectroscopy now provides much easier manipulation of the cells of interest and 10 times higher throughput.

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

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

Pick a cell with negative pressure, measure, release it again with a positive pressure pulse or by a short cleaning procedure. Image Credit: Cytosurge AG

For Cell Mechanobiology & Biophysics

The physical study of single cells enables insights into biological phenomena like the differentiation, growth and proliferation both in biophysics and mechanobiology. In stem cells, cancer research, and organoids, the cell mechanical properties and interactions with its environment are key to gaining a better understanding.

Additionally, for implant materials there is a clinical requirement to understand and control how various cells adhere. One aim is to understand these influences on a single cell level, as each cell is different from its neighbor (also known as cell heterogeneity).

Single cell force spectroscopy has been established as an insightful technique which can be used to address such questions by utilizing atomic force microscopy (AFM). However, the process of gluing a cell to an AFM cantilever is time-consuming and tedious, limiting the throughput to just a few cells per day.

Yet, multiple cells are usually required to meaningfully examine any experimental condition because of cell-cell heterogeneity. Here, by reversibly immobilizing a cell to a FluidFM probe by suction, and subsequent release with pressure, FluidFM supplies a dramatic improvement. This enables the increase of the throughput by a factor of 10 to 100, and so the ability to acquire sound statistics within one day instead of weeks or months.

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

Measure Suspended or Fully Adherent Cell

FluidFM micropipettes enable the user to either directly pick up a suspended cell from a fully adherent culture, or to attract it from solution. FluidFM is able to overcome cell adhesion forces of over 1000 nN.

Largest Force Range, Highest Flexibility

Reusable measurement probes in combination with reduced preparation time makes FluidFM the ideal tool for all single cell mechanical studies. Due to the unique properties of FluidFM technology, the user is able to collect solid cell mechanical data in much less time. In addition to gaining access to unparalleled measurement ranges and increasing experimental flexibility.

From pN to µN

Take advantage of FluidFM probes with different stiffnesses and opening diameters to measure forces from tens of pN up to µN. With available openings from 300 nm to 8 µm and spring constants from 0.3 N/m to 4 N/m, a large range of cells and mechanics can be covered.

Pick. Measure. Clean. Repeat.

Many mammalian cells have a tendency to quickly adhere to any substrate, as well as the FluidFM probe, cleaning the probe after each cell is a common approach. The procedure takes under two minutes before picking up the next cell for measurements. This cleaning procedure may be skipped for non-adherent cells, and the throughput is even higher.

Illustration of a cell being detached from a substrate by FluidFM. The measured force-distance curve can give insights on adhesion strength, energy, distance as well as the involved bio-chemical bonds.

Illustration of a cell being detached from a substrate by FluidFM. The measured force-distance curve can give insights on adhesion strength, energy, distance as well as the involved bio-chemical bonds. Image Credit: Cytosurge AG

Publication Highlights

FluidFM has been employed in a number of publications in order to quantify cell mechanical properties. Three highlights are presented below.

Optimizing Drugs

Millions of people around the world suffer from Leukemia. Although treatment drugs exist, cancer regularly develops a resistance against them. With the help of FluidFM cell measurements, researchers from both the University Würzburg and University Hospital Würzburg found a new potential method to overcome resistance to the recently approved Midostaurin drug and even heighten the drug activity.

A. Garitano-Trojaola, et al..  RAC1 Inhibitor EHT1864 and Venetoclax overcome Midostaurin resistance in Acute Myeloid Leukemia. (2019) Blood.

Optimizing Stents

Stents save many lives by helping millions of people every year overcome arterial blockages. Once it is implanted, the stent should integrate well and stop the formation of blood clots. In this article research groups from ETH Zurich examine stent design optimization by measuring cell adhesion to its surface using FluidFM.

E. Potthoff, et al. Toward a rational design of surface textures promoting endothelialization. (2014) Nano Letters, 14(2), 1069-1079.

Calibrating High-Throughput Devices

Single cell force spectroscopy provides fundamental insights into multiple fields, yet it suffers from low throughput. MTA Budapest speeds up the acquisition of single cell adhesion data massively, by utilizing FluidFM adhesion measurements to calibrate an optical sensor array. So, they are able to monitor over 1000 adherent cells mechanically, in parallel.

M. Sztilkovics, et al. Single-cell adhesion force kinetics of cell populations from combined label-free optical biosensor and robotic fluidic force microscopy. (2020) Scientific Reports.

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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