Acoustic Force Spectroscopy

Acoustic Force Spectroscopy (AFS­™) is LUMICKS’ highly parallel single-molecule manipulation method, capable of applying forces on thousands of biomolecules in parallel with high precision.


Acoustic Force Spectroscopy

The essence of the AFS™ technology lies in a glass microfluidic chip with two piezo elements that generate resonant acoustic waves (ultrasound). These resonant acoustic waves are used to exert forces on micrometer sized particles with a different density than the surrounding medium (e.g. a polystyrene microsphere or a living cell). Explore the applications below to see what this technology can mean for you.

In the compact AFS™ chip the acoustic waves are employed to exert forces from sub-pN to hundreds of pNs on thousands of biomolecules (such as DNA, RNA or proteins) or cells in parallel, with sub-millisecond response time and inherent stability.

acoustic-force-spectroscopy-explanation

Further reading:
Sitters et al. (2015) Acoustic Force Spectroscopy. Nature Methods

Applications

AFS™ enables scientists to investigate the mechanical properties of biological systems by probing them via synthetic microspheres or by directly probing the system itself (e.g. a cell). Due to the planar nature of the acoustic waves, experiments can be performed in a highly parallel fashion. This makes AFS™ an ideal tool to study a wide variety of biological systems, including, but not limited to: DNA manipulation, Antibody – antigen affinity, protein unfolding, cell manipulation, microrheology, lipid membrane interactions, enzymatic activity and DNA-protein interactions.

Measure Protein Unfolding

Measure Enzymatic Activity

Measure DNA-Protein Interactions

Force Extension and Manipulation of DNA

Measure Antibody-Antigen Affinity

cell-manipulation-AFS-thumb

Acoustic Force Cytometry

AFS™ Stand-alone

AFS™ Stand-alone is LUMICKS’ true turnkey single-molecule tool. It includes a dedicated inverted optical microscope and a powerful workstation with a LabView-based software package. With the AFS™ Stand-alone you will be able to focus on your experiments right away, without the need for additional software or imaging tools.

Acoustic Forces

• Range from 0 to 50 pN* (4.5 µm diameter polystyrene microspheres)
• Static and dynamic force measurements with 14 bit resolution
• Loading rates ranging from 10-4 pN/s to 103 pN/s

* Upgradable to 200 pN using external amplifier, includes bypass feature

Workstation & Software

Powerful workstation including LabView-based open-source software capable of:
• Real-time parallel 2D tracking of 1500 microspheres at 25 Hz
• Real-time parallel 3D tracking of 400 microspheres at 25 Hz
• Real-time position accuracy of 2 nm (XY) and 4 nm (Z) at 25 Hz

Imaging

USB 3.0 CMOS camera* with:
• Resolution of 1.3 MP
• Real-time parallel measurements up to 60 fps in full field of view

* Upgradable to camera with 2.3 MP and 166 fps

acoustic-force-spectroscopy-stand-alone

AFS™ Module

In addition to the AFS™ Stand-alone microscope platform, LUMICKS offers the AFS™ Module for use with commercial inverted optical microscopes. This combination grants you the ability to study mechanical manipulations of biomolecules and cells with the powerful visualization capabilities of state-of-the-art fluorescence and super-resolution microscopy.

Acoustic Forces

• Range from 0 to 200 pN (4.5 µm diameter polystyrene microspheres)
• Static and dynamic force measurements with 14 bit resolution
• Loading rates ranging from 10-4 pN/s to 103 pN/s

Software

Open-source software (LabView-based) capable of:
• Constant forces
• Dynamic force ramps
• User-defined time varying force profiles

The AFS™ Module software does not provide image analysis tools

acoustic-force-spectroscopy-module

Highlighted Features

The AFS instrument is a versatile tool which can be used to answer a broad spectrum of research questions. Here we highlight two enabling features and their applications.


Imaging Possibilities

40x-Objective-Field-of-View acoustic force spectrometry

40x CFI Plan Achromat

Field of view: 284 x 178 μm

This 40x magnification lens is perfect for use with small particles or if a high tracking accuracy is required.

20x-Objective-Field-of-View

20x CFI Plan Fluor

Field of view: 568 x 356 μm

The 20x objective provides a nice balance between parallelism and accuracy.

10x-Objective-Field-of-View

10x CFI Plan Fluor

Field of view: 1136 x 712 μm

The 10x magnification is a great pick for boosting the high-throughput ability of AFS.

Bead size = 4.5 μm. Included in AFS™ Stand-Alone

 

Temperature Control

The temperature controlled AFS chip holder is designed to allow users to manipulate cells or biomolecules under biologically-relevant conditions and investigate temperature-dependent events in single-molecule experiments. The AFS chip holder is made from heat-conducting materials and contains two heating elements that can reach the desired temperature with an accuracy of 0.4º C. The temperature controlled chip holder is compatible with both the AFS Stand-Alone and the AFS module.

Specifications Overview

A detailed overview of options & specifications

Features AFS Stand-alone AFS Module AFS Module with integration package (tracking software, z-stage, illumination, camera)
XY-Tracking
Z-Tracking
Force calibration
Inverted fluorescence microscopy compatibility  ●*
 

Applications

Force-distance curves
Bond ruptures
Force application

 

* If the AFS chip holder is integrated on an inverted (fluorescence) microscope the same functionalities and applications as the AFS Module apply.

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