Acquiring 4D confocal quality images can be difficult with some types of samples and experiments. If the fluorescence signal is weak or very photo-sensitive and the sample is quickly damaged or even killed by UV light, using a laser based imaging system is not an option. Achieving results in these circumstances can be time consuming and frustrating, but now there is an alternative that requires minimal sample exposure, yet produces exceptional images quickly and easily.

Using the Volocity Restoration Confocal, you can produce confocal quality images in real time, from a standard research microscope, without using lasers or adding devices to your system. Images are acquired at intervals in Z through the sample and the powerful Volocity Restoration algorithms deconvolve the whole volume of data to produce confocal images. This means that sample exposure can be kept to a minimum as the entire emission signal is collected and nothing is discarded during acquisition. Time lapse experiments may last longer as the fluorochrome is not bleached and cells do not suffer from photo-toxic effects.

Drosphila germarium labeled for the cohesive protein ORD, before and after deconvolution using Volocity Restoration.

The system includes two algorithms for image restoration; which one you choose to use depends on the type of experiment you want to perform and how you wish to use the end result. Online deconvolution is achieved using the Fast Restoration algorithm, an ultra-fast routine that uniquely uses every voxel in the volume in a single pass process. This feature is very rapidly computed and produces superior results when viewed in XY. Fast Restoration is perfect for improving the visual quality of your images in real time.

For a result that improves the Z resolution of the image volume as well as the resolution in X and Y, the Iterative Restoration algorithm will produce superior results. Iterative Restoration includes an award winning restoration algorithm developed by Improvision from published Maximum Entropy techniques. It uses a restorative technique that reassigns out-of-focus haze rather than subtracting it from the data. Iterative Restoration is more computationally intense process and therefore takes longer to produce results than Fast Restoration, so is used post-acquisition. For convenience the algorithm can be applied as a batch process across multiple volumes. The resulting images can be used for measuring fluorescence intensity as well as for morphological analysis.

Image capture is controlled by Volocity Acquisition. The award winning design of the user interface makes the acquisition dialog easy to configure - from 2D, single channel capture to complex time resolved, 3D multi-channel experiments. Include a scanning XY stage in your hardware and you can perform multi-point image acquisition on a single slide or multi-well plate, capturing in 2D, 3D or 4D from every point or well.

The Volocity Restoration Confocal includes Volocity Visualization for fully interactive volume rendering of 3D and 4D data sets. Channel opacity and brightness can be individually controlled so that you can achieve the view you need. Play through time resolved 3D volumes so that you can relate structure to function whilst interacting with the volume, for a better understanding of your results. Volocity Visualization provides the tools to create publication ready images and movies.

The Volocity Restoration Confocal system is part of the Volocity product family so can easily be extended to include features for object detection, measurement, tracking, colocalization and other applications.

If you need to produce confocal quality images with minimal photo-bleaching, the Volocity Restoration Confocal may be the solution for you. Please contact us for more details.