Volocity FRAP Plugin

Fluorescence recovery after photobleaching (FRAP) is an imaging technique which is widely used by life scientists to study the mobility of molecules in cells. The Volocity FRAP Plugin provides an easy to use, reliable method for FRAP imaging. Image acquisition, data analysis and charting all take place using a seamless workflow through the user-friendly Volocity interface.

The Volocity FRAP Plugin provides all the tools needed to successfully acquire FRAP images, but unlike some FRAP solutions, it doesn’t stop there. What sets this product apart is the powerful and rapid way in which the FRAP data is analyzed and displayed.

During a FRAP experiment, a number of image sequences (typically 10) are acquired. Following acquisition, corrections for background and photobleaching are quick to perform by applying to all the sequences in the experiment using batch processing. You can then choose which equation of best fit to apply to the data; single exponential, double exponential and the Soumpasis analytical solution are included. For convenience, the data is automatically normalized by the software so that the pre-bleach intensity is set to 100 and the t0 intensity is set to 1, which means that all the results from all the data sets can be pooled for one experiment.

The Volocity FRAP Plugin will then instantly display your FRAP data in the form of a chart, by fitting the data to the equation you have chosen to use. Not only is the data charted, the software also calculates and displays the important values for the mobile fraction (Fm), the immobile fraction (Fi) and the T 1/2. The chart can be exported for inclusion in reports, along with the image data. To see the numbers used in the chart, simply click the "Raw" tab – you can export the data from here too should you wish to do so.

Follow the workflow below to see how easy and powerful the Volocity FRAP Plugin is to use. A range of bleaching devices is supported, see the Volocity Supported Hardware List for details.

Step 1. Configure the imaging protocol using the FRAP acquisition setup dialog.

Step 2. Set the ROI on the FRAP preview.

Step 3. Acquire FRAP image data. Apply background correction and photobleaching correction by batch process.

Step 4. Choose the equation of best fit from the FRAP Analysis dialog.

Step 5. Click on the "Analyse" button and the data is almost instantaneously displayed as a chart. See the data by clicking the "Raw" button.

Requires Volocity Acquisition and Volocity Quantitation. With the appropriate laser configuration, the setup can be also used for iFRAP, FRAP/FLIP, photoactivation and uncaging. This product is only available with the Volocity Spinning Disk Confocal at the present time.

For more information on the Volocity FRAP Plugin, please use the Contact form.