The atomic force microscope that combines versatility and performance for biology and life science
Compatible with inverted microscopes Flat and linear scanning thanks to flexure-based scanner technology True flexibility with exchangeable cantilever holders for specialized tasks More measurement versatility with the FlexAFM’s scanning capabilities in liquid and its additional measurement modes
A key success factor in life science research is the combination of multiple techniques. With the Flex-Bio, Nanosurf's Bio AFM, you can combine the liquid AFM imaging, spectroscopy and nanomanipulation capacity of this system with the high-end optical techniques available for inverted microscopes.
Flexible system design for life science research
Flex-Bio comes with manual and motorized stages for seamless integration on Zeiss, Olympus, Nikon and Leica inverted microscopes or with standalone stages. On the inverted microscope, optical and AFM data can be correlated, as shown here for internal limiting membrane (ILM) of the human retina.
(A) Bright field image of isolated ILM in a physiological buffer. (B) Fluorescence image of the same section showing anti-laminin staining. (C) AFM topograph of a subsection of the ILM; also shown as overlay in B. (D) AFM stiffness measurements (stiffness map) of the same subsection. The color for each point represents the local stiffness value as calculated from force curves recorded at the respective positions. (E) Histogram of the stiffness data shown in D. (F) Typical force-displacement curves obtained on the ILM and on the glass substrate. These curves are converted to force-indentation data, which then allows calculation of the stiffness. Stiffness distribution of biological tissues has been shown to be a marker for diseases such as age-related macular degeneration, arthritis and cancer. Data courtesy: Marko Loparic, Marija Plodinec, Philip Oertle, and Paul B. Henrich, Biozentrum/SNI/UHBS, University of Basel, CH.
The modular stage, cantilever holder, and software concept allows an easy upgrade of the system to access many new possibilities in life science and materials research. Flex-FPM for cell and nano-manipulation, for example, and Flex-ANA for automatic nanomechanical analysis. In addition, advanced modes like MFM and KPFM that were originally developed for the Flex-Axiom system, are also available for Flex-Bio. For measurements that don’t need optical access from below, e.g. for the imaging and spectroscopy of samples like bacteriorhodopsin, a standalone stage makes the Flex-Bio compatible with the Nanosurf Isostage and Acoustic Enclosure 300, and generally makes the system much more compact.
A Flex-Bio system with stand-alone stage, isostage, and acoustic enclosure. (B) 2D crystals of bacteriorhodopsin [140 nm scan range]. (C) Power spectrum of B, showing a lateral resolution of well over 1 nm [dashed circle]. (D) Single molecule force spectroscopy of bacteriorhodopsin.
Practical details that really matter in daily use
Cantilever holders with alignment structures are available for use with cantilevers containing alignment grooves. This provides micrometer repositioning accuracy, circumventing laser alignment and allowing you to find the same sample features again and again. Cantilevers enter the image from top to bottom, so sample orientation is always the same, no matter whether you look by eye, CCD camera, or AFM (when scanned at the default scan angle).
Alignment chip technology
Top view on stand-alone stage
Top view on inverted microscope, image courtesy: O. Guilaume-Gentil, ETH Zürich, Switzerland
AFM Modes
Flex-Bio imaging modes
This overview shows which modes the instrument is capable of. Some modes may require additional components or software options. For details, please view the brochure or contact us directly.
Standard imaging modes
Static Force Mode
Lateral Force Mode
Dynamic Force Mode (Tapping Mode)
Phase Imaging Mode
Thermal imaging modes
Scanning Thermal Microscopy (SThM)
Magnetic properties
Magnetic Force Microscopy
Electrical properties
Conductive AFM (C-AFM)
Piezoelectric Force Microscopy (PFM)
Electrostatic Force Microscopy (EFM)
Kelvin Probe Force Microscopy (KPFM)
Scanning Spreading Resistance Microscopy (SSRM)
Mechanical properties
Force Modulation
Stiffness and Modulus
Force Spectroscopy
Force Mapping
Other measurement modes
Lithography and Nanomanipulation
Electrochemical AFM (EC-AFM)
Applications
Flex-Bio application examples
Imaging of type I collagen fibrils
Collagen is the most abundant protein in mammals and contributes to more than 25% of the whole-body protein content. It is the main structural protein of the extracellular matrix of connective tissues and provides e.g. tendons and bone with their tensile strength. Most of the collagen found in mammals is fibrillar type I collagen. Type I collagen fibrils show a typical periodic morphology, the so-called D-banding. D-bands result from staggered self-assembly of individual collagen molecules into larger fibrils with a periodicity of about 67 nm.
Images of collagen fibrils from rat tendon were recorded in Prof. Snedeker's research group at the ETH Zürich. One of the reasearch areas of Prof. Snedeker is tendon mechanics and biology.
3D AFM topography of several type I collagen fibrils
AFM topography image of type I collagen fibrils
AFM deflection image recorded along with the topography image
The 3D representation of the AFM topography image nicely shows the typical periodic D-banding of type I collagen on all fibrils. The colloagen topography was recorded in static mode using a Nanosensors PPP-XYCONTR cantilever. AFM images were processed using Nanosurf Report Software. Preparation and imaging of collagen fibrils was performed by Massimo Bagnani, Prof. Snedeker research group, Uniklinik Balgrist, Institute for Biomechanics, ETH Zürich, Switzerland.
Measurements on living cultured cells
Mechanobiology is an emerging research area that deals with the effect of changing physical forces or changes in the mechanical properties of cells and tissues. Several diseases, such as fibrosis and atherosclerosis are associated with changes in tissue stiffness. Moreover, in cancer, the metastatic potential of cancer cells depends on their elastic modulus.
Here, the elastic modulus of living cells from a human breast basal epithelial cell line was measured using a Nanosurf Flex-Bio system with the Flex-ANA software.
Elastic modulus map
Unperturbed cell topography from force mapping
The first image shows the elastic modulus (in kPa) recorded on living breast epithelial cells immersed in cell culture medium. Differences in the elastic modulus within the cell can be clearly observed. The dark area surrounding the cells originate from the much stiffer cell culture dish substrate.
The second image shows the unperturbed cell topography extracted from the force mapping data. The topography is determined from the contact point of each force curve and thus shows the cell topography at zero applied force.
Elastic modulus mapped to the 3D topography
Elstic modulus distribution
Mapping the elastic modulus data to the 3D topography allows relating the information of both channels. The 3D image was generated using Gwyddion software.
The last image shows the distribution of the elastic moduli extracted from nanomechanical force mapping experiments. The peak at lower moduli corresponds to the stiffness of the cells. The peak at the right originates from the cell culture substrate and shows much higher elastic moduli.
AFM data courtesy of Philipp Oertle, Biozentrum, University Basel.
Single molecule force spectroscopy of bacteriorhodopsin
The force-distance curve below reports the controlled C-terminal unfolding of a single bacteriorhodopsin (BR) membrane protein from its native environment, the purple membrane from Halobacterium salinarium.
Solid and dashed orange lines represent the WLC curves corresponding to the major and minor unfolding peaks observed upon unfolding BR, respectively. The contour length of the stretched polypeptides of the major unfolding peaks is given in amino acids (aa).
Single molecule force spectroscopy of bacteriorhodopsin
This data was recorded using a FlexAFM scan head (10-µm; version 3) in combination with the C3000 controller and a cantilever with a nominal spring constant of 0.1 N/m (Uniqprobe, qp-CONT, Nanosensors).
Options & Accessories
Flex-Bio options and accessories
At the heart of the Flex-Bio system are the FlexAFM scan head and the C3000 controller with I100 interface box. On top of that, several options and accessories are available, designed to deliver stable and reliable results, or to provide access to new measurement possibilities and experiments for this Bio AFM.
Cover slip holder with heating option
Environmental control chambers from several inverted microscope providers and Life Imaging Services (LIS) to support AFM cell imaging under conditions favorable to your cells
Vibration isolation
Vibration-free measurements with the Halcyonics_i4
State-of-the-art-active vibration isolation system
Ideal isolation from building vibrations and other disturbances
Isolation effect starts at 0.6 Hz and achieves max. performance of –40 dB at 10 Hz, where 99.0% of the vibration is isolated
Acoustic enclosure
Protection from airborne noise
Protect your equipment from airborne noise emitted by air conditioning, venting, door slamming etc.
Enables you to perform undisturbed experiments with your Flex-Bio setup.
Isostage 300
Vibration-free measurements with the Nanosurf Isostage. Requires an adapter. Only suitable for use without inverted microscope.
Isostage adapter plate
FlexAFM sample stage with sample holder
Sample plate with 120 mm diameter
Fits on Isostage 300
Flex-ANA upgrade
Nanomechanical tissue diagnostics and soft material analysis
Fully automated measurements on rough and non-even surfaces
Quantitative analysis of tissues and soft materials alike
Fast, objective, and routine sample categorization
FluidFM® upgrade
An enabling technology for micro-manipulation of single cells and other small objects, surfaces and tissues
Nanofluidics through a hollow cantilever combined with the positional accuracy and force control of the Nanosurf FlexAFM
Specialized application modules for different applications as injection, pick-and-place, adhesion force spectroscopy, elasticity measurements, and spotting
Suitable for stand-alone setup and on the inverted microscope
Specifications
Flex-Bio specifications and dimensions
System specifications
FlexAFM NIR scan head specifications
NIR 110-µm
NIR 10-µm
Laser class (wavelength)
Class 1M laser product (850 nm)
Maximum Petri dish height (fluid level)
9 mm (6 mm)
Manual approach range
30 mm
Automatic approach range
1.1 mm
Maximum scan range
100 µm(1)
10 µm(1)
Maximum Z-range
10 µm(2)
3 µm(1)
Drive resolution in XY
0.006 nm(3)
0.0006 nm(3)
Drive resolution in Z
0.0006 nm(3)
0.0002 nm(3)
XY-linearity mean error
< 0.1 %
XY-flatness at maximum scan range
typ. 5 nm
typ. 1 nm
Z-measurement noise level (RMS, dynamic mode in air)
typ. 0.03 nm
Scan head dimensions
143 x 158 x 53 mm
Scan head weight
1.25 kg
(1) Manufacturing tolerances ± 5%
(2) Manufacturing tolerances ± 10%
(3) Maximum theoretical resolution; calculated by dividing the maximum range by 24 bits
C3000i controller — Core hardware specifications
X/Y/Z-axis scan and position controller
3× 24-bit DAC (200 kHz)
X/Y/Z-axis position measurement
1× 24-bit ADC (200 kHz)
Excitation & modulation outputs
2× 16-bit DAC (20 MHz)
Analog signal input bandwidth
0–5 MHz
Main input signal capturing
2× 16-bit ADC (20 MHz)
2× 24-bit ADC (200 kHz)
Additional user signal outputs
1× 24-bit DAC (200 kHz)
Digital synchronization
Sync Out 1/2: digital outputs, signal range 0/5V TTL pulses
FPGA module and embedded processor
ALTERA FPGA,
32-bit NIOS-CPU,
80 MHz, 256 MB RAM,
multitasking OS
Communication
USB 2.0 Hi-Speed to PC and scan head interface
System clock
Internal quarts (10 MHz) or external clock
Power
90–240 V AC, 70 W, 50/60Hz
Cantilever
Width
min. 20 μm
Length
min. 40 μm
Reflective coating
Reflective coating recommended
Liquid measurements
Yes, with gold coating
Alignment grooves
Required by default
Special cantilever holders without alignment grooves are available
Resonance frequency dynamic mode Easyscan 2 Controller
15 kHz to 350 kHz
Resonance frequency dynamic mode C3000i Controller
< 4 MHz
Cantilever shape
Single rectangular cantilevers and multilever cantilevers (depending on scan head version and cantilever holder)
Chip thickness
300 μm, 500 μm or 600 μm depending on cantilever holder
Scan head dimensions
Software
Control software for Flex-Bio
The control software for Nanosurf AFMs is an intuitive platform made for performing your AFM measurements efficiently and easily. Our Service team and software engineers are constantly developing and implementing new features and enhancements to further improve the user experience. We regularly publish new and improved versions, which you can download for free. You can install the software on as many computers as you wish to analyze your data.
Free lifetime updates: download all software updates for free
All software updates for Nanosurf control software are free of charge. Our software team is constantly working on new features and improvements to make the user experience better, more intuitive and more efficient.
Software features
Automatic/parameter-free frequency tuning based on cantilever characteristics
Simply choose the cantilever you are using, and the system automatically performs the frequency sweep prior to approaching the sample. No manual setting of parameters is required.
Distance measuring tools: measure the distance between points or lines, the height of features, and more
A selection of different measuring tools allow you to accurately measure angles and distances directly on the acquired measurement image.
Determine the distance between two points or between two parallel lines to make very precise measurement (as shown in the video).
Integrated stage control for various motorized stages
The software lets you control your motorized stage out of the box. Load your stage's configuration file and you are ready to move the stage out of the software, either using arrow buttons, or by entering precise cantilever positions on the sample.
More convenient usability features:
Spectroscopy wizard: follow easy steps to set up spectroscopy measurements
One software UI for all scan heads: no additional learning curve if you use multiple Nanosurf AFM systems
Automated deflection calibration
UI layouts for beginners and advanced users
Highly configurable graph area with mode-dependent auto-layout: the software automatically shows the relevant graphs and information
Easy file handling with comfort features: auto apply naming conventions, Windows Explorer integration, image gallery, bulk renaming.
Includes a powerful scripting interface: automate and extend capabilities according to your needs. Compatible with most programming environments (e.g. LabView, Python, MatLab, C++, Java, and more)
