3D Targeted Illumination - 3D FRAP

The release of Volocity 5.2 pioneers new dimensions in Photokinesis and brings you the world's first complete acquisition to analysis 3D targeted Illumination solution. The photokinesis unit is an essential accessory for scientists who wish to perform Fluorescence Recovery After Photobleaching (FRAP), Fluorescence Loss In Photobleaching (FLIP), photoswitching (photoconversion) and acceptor photobleaching.

Fluorescence Recovery After Photobleaching

FRAP exploits the photobleaching characteristics of fluorophores for measuring the mobility of fluorescently-labeled proteins at a sub-cellular level. With FRAP, the fluorescently-tagged protein of interest is visualized at low light intensity followed by photobleaching of a user-defined region of interest with high intensity light, causing the fluorophores to bleach in the selected region.

Corrections for background and photobleaching can be performed rapidly in Volocity by applying to all the sequences in the experiment using batch processing. You can then choose which equation to apply to the data.  Single exponential, double exponential and the Soumpasis analytical solution are included.

FRAP data is displayed in the form of a chart, by fitting the data to the equation you have chosen to use. Not only is the data charted, but Volocity also calculates and displays the important values for the mobile fraction (Fm), the immobile fraction (Fi), the diffusion co-efficient and the T 1/2. The chart can be exported for inclusion in reports, along with the image data.

3D Fluorescence Recovery After Photobleaching allows the recovery to be visualized and measured in 3D space to offer new experimental possibilities. Bleaching is performed as part of an acquisition protocol so that pre- and post-bleach volumes are acquired. Targeting a specific plane in your 3D volume for high intensity illumination allows you to observe the post-bleaching of the 3D stack which has never before been possible. This new technology will allow researchers to create pioneering new experiments and increase the understanding of your life science.

3D Targeted Illumination

Volocity software updates for scientific imaging are produced to the highest professional coding standards, with user interfaces which are attractive and easy to use. Our team welcomes all suggestions from our users for incremental improvements and major new features which help to keep our customers at the forefront of technology and to be leaders in their life science research.

Volocity 5.2 features a range of new advancements driven by the scientific imaging community offering you even more flexibility to perform the exact experiments you require. 

Key Features

• Pre-bleach image acquisition phase providing images for normalization.
• Recording of bleach timing of metadata for accurate analysis of recovery.
• Choice of single or multiple user defined bleach regions within one experiment.
• Choice of analysis equations for pure diffusion, single or multiple binding states.
• Live charting of recovery allows user to determine experimental end-point accurately.
• Automatic recording of bleached regions and their metadata to allow accurate analysis.
• Automatic saving of all images and metadata to allow analysis and reanalysis within Volocity.
• Post-bleach image acquisition can be divided into phases with separate capture rates for better optimization.

To learn more about 3D targeted illumination in Volocity and for a full list of technological advancements now available in Volocity 5.2 please contact us.