Mechanobiology with FluidFM®
This application note introduces the basic principle of mechanobiology and notably the use of the FluidFM technology for mechanobiology applications.
Go straight to: Definition | Single-cell force spectroscopy with the FluidFM technology | Applications
What is mechanobiology?
Mechanobiology - the definition
The term "mechanobiology" refers to "A field at the interface of biology, physics, and bioengineering, which focuses on how cell/tissue mechanics and physical forces influence cell behavior, cell and tissue morphogenesis, and diseases related to these processes." [1]
Why is mechanobiology so important?
Cells feel and respond to various mechanical signals from their surroundings. The mechanical stiffness of the extracellular matrix strongly determines cell function, stem cell differentiation and tissue homeostasis [2-6]. Oppositely, changes in the matrix stiffness can support the development and spreading of diseases, such as cancer and fibrosis [4]. In addition, cells are often subjected to shear stress due to several physiological processes. In a nutshell, cells are highly sensitive to external forces. Given their central importance in cell function and human health, the literature accumulated has showed that these forces are ubiquitous in vivo. As a result, mechanobiology has emerged as a new and growing field that attracts researchers from various disciplines. The field of mechanobiology comprises a range of field of applications and tools to perform force measurements between various systems and in different environments (air, liquid).
How does mechanobiology with FluidFM work?
Mechanobiology applied - single-cell force spectroscopy with the FluidFM technology
Traditionally, in
Single-Cell Force Spectroscopy
(SCFS) assays, the object of interest is glued to an AFM cantilever resulting in complex handling and low throughput.
The FluidFM systems solve this issue by
reversibly immobilizing a cell to a FluidFM probe
via suction, and subsequent release with a pressure pulse or brief washing.
This gentle exchange of the cell allows the cantilever to be re-used for several measurements, saves time and costs, and results in a 10 times higher throughput compared to traditional methods. Consequently, both throughput and efficiency of measurements are increased, drastically reducing the time required to obtain statistically relevant data compared to conventional methods. [7] Hence, this technology broadens the possibilities for highly reproducible single-cell adhesion measurements for a broad range of applications.
Measure up to 200 cells a day
High throughput obtained with a simple and reversible cell immobilization
Broad force range
Direct and semi-automated force measurement from pN up to µN
Many cell types & colloids
For mammalian cells, microbes, and colloids
Advantages of using FluidFM for mechanobiology
Speed-up your mechanobiology research with a semi-automated single-cell adhesion measurement
The FluidFM OMNIUM is a semi-automated system for measuring up to 200 single cells a day. Get reproducible, direct force measurements in high quality and with sound statistics.
High throughput
Direct force measurement
Easy-to-use & reproducible
Compatible with standard cell-laboratory materials
10x faster measurements compared to standard methods
As objects like cells or colloids can be quickly exchanged through reversible immobilization, measurement throughput is increased more than 10-fold. Up to 200 individual objects can be analyzed in a single day.
Working principle of single cell force spectroscopy with FluidFM.
Simple - no glue needed
The suction method of immobilizing the objects onto the FluidFM cantilever makes it reversible and avoids any glue: Pick. Measure. Release. Repeat.
Fast & easy. In this video, three micrometer colloids are attracted from suspension with a vacuum, held briefly, and then released again with a pressure pulse.
Switch the probe anytime or reuse to save money
Whether due to degradation, contamination or a required change of probe geometry or chemistry – you can switch the probe at anytime. Just release the object, change the probe, and take up the object again. Our FluidFM Probes typically last for several force spectroscopy measurement days allowing the analysis of several hundred cells.
Many cell types and colloids supported
Cells and colloids come in a wealth of shapes and sizes. FluidFM single-cell force spectroscopy works with adherent or suspension mammalian cells, spheric or rod-shaped microbes, and with colloids, bubbles, and droplets from 0.5 to 100 µm particle size. FluidFM can handle them whether they are hundreds of nm or dozens of µm in diameter. Customers have even analyzed non-colloidal E.coli cells.
| Cells | Microbes | Colloids |
|
|
|
|
| Adherent or suspension cells | Spheric or rod shaped. Algae, bacteria, protozoa, and fungi. |
Colloids from 0.5 to 100 µm particle size. Also for bubbles, droplets. |
Self-centering – means reproducibility
The position of the object on the FluidFM cantilever is given by the position of the aperture. Thus, every colloidal probe will be centered automatically and at the same position – as long as the same FluidFM Probe is used. This results in highly reproducible positioning.
1) cell is selected 2) Cell is detached from surface 3) Resulting force spectroscopy. Image courtesy of Bruker.
10x higher force range
The various stiffnesses and opening diameters of FluidFM Probes enable to measure forces from tens of pN up to µN.
Pick from substrate or attract from solution, or even air
Pick-up cells directly from a substrate or attract them from a solution via liquid influx to the aperture of the FluidFM Probe. This method is also recommended when the long-term adhesion of a microbe to a substrate is too strong to quantify, and hence shorter-term interactions are studied. Researchers have also performed particle and microbe measurements in air.
S. Cerevisiae, also known as baker’s yeast, are picked-up from medium, measured and then deposited in a line with a FluidFM Micropipette. The cells stay fully viable through this procedure. Image courtesy of P. Dörig, ETH Zurich.
Which FluidFM system suits your application?
Mechanobiology applications with the FluidFM technology
Study of tumor progression and metastasis with cell-cell adhesion forces
Study with a Nanosurf Flex-FPM by Dr. Noa Cohen, group of Prof. Tanya Konry, Northeastern University in Boston, on cell-cell adhesion forces to gain more insights into tumor progression and metastasis (Cohen et al., 2017):
Optical images showing:
A) a single cell to be picked up by a FluidFM Probe
B) the cell aspired to the cantilever and
C) the FluidFM Probe with aspired cell during a cell-cell adhesion measurement.
Data/image courtesy of Tanya Konry group, Northeastern University, Boston, USA.
A) Typical force curves between a MCF7 cell aspired to the cantilever and non-cancerous, fibroblast (HS5) on the substrate at different contact times.
B) Development of the force with contact time between the cells.
Data/image courtesy of Tanya Konry group, Northeastern University, Boston, USA.
References
[1] Jansen, Karin A., et al. "A guide to mechanobiology: Where biology and physics meet." Biochimica et Biophysica Acta (BBA)-Molecular Cell Research 1853.11 (2015): 3043-3052.
[2] Engler, Adam J., et al. "Matrix elasticity directs stem cell lineage specification." Cell 126.4 (2006): 677-689.
[3] Orr, A. Wayne, et al. "Mechanisms of mechanotransduction." Developmental cell 10.1 (2006): 11-20.
[4] Lu, Pengfei, Valerie M. Weaver, and Zena Werb. "The extracellular matrix: a dynamic niche in cancer progression." Journal of cell biology 196.4 (2012): 395-406.
[5] Eyckmans, Jeroen, et al. "A hitchhiker's guide to mechanobiology." Developmental cell 21.1 (2011): 35-47.
[6] Wang, J. H-C., and B. P. Thampatty. "An introductory review of cell mechanobiology." Biomechanics and modeling in mechanobiology 5.1 (2006): 1-16.
[7] Dehullu, Jérôme, et al. "Fluidic force microscopy captures amyloid bonds between microbial cells." Trends in microbiology 27.9 (2019): 728-730.