Olympus cellFRAP is designed with researchers in mind, providing highly accurate and flexible live-cell photomanipulation with various evaluation options for data presentation needs. cellFRAP employs a diffraction-limited laser spot for high-intensity and precise positioning of the bleaching region across the full field of view. The systems’ excellent spot quality allows full flexibility in position and shape of the bleaching region, and enables control of the bleaching area with extreme accuracy. Via incorporation into the Olympus IX3 open source frames, the cellFRAP deck can easily be adapted to an existing imaging platform with a full range of compatible accessories. Using the cellFRAP solution in the Olympus cellSens Graphical Experiment Manager provides simple system set-up and experiment control.

Highest Spatial and Temporal Resolution
The cellFRAP system guarantees highest spatial and temporal resolution for acquiring reliable and reproducible data in photomanipulation applications. Based on the system’s diffraction-limited spot properties, bleaching areas can be controlled extremely accurately. Driven by the Olympus Real-time controller, the cellFRAP system offers an unrivaled short delay time of only 200 µs between bleach and postbleach acquisition. Therefore, the system enables observation and collection of all important and immediate sample responses.

Diffraction-Limited Laser Spot
The Olympus cellFRAP solution enables the best photomanipulation results without the need to move the sample. By utilizing a diffraction-limited laser spot, cellFRAP does not only guarantee the highest bleaching intensity but also ensures a precise and flexible choice in shape and position of the bleaching regions across the full field of view. High-quality chromatically corrected optics open up the possibility of using up to four different lasers during one experiment—for example, for optogenetics applications.

Reproducibility
Ultraprecise experimental control and reproducibility are of interest to most users. The cellFRAP system accounts for these demands by ultraprecise experimental control given by high system-level synchronization at ~1 µs temporal resolution. In addition, the complete workflow of an experiment is documented and can easily reproduced at any time.

Straight Forward Setup and Control
Working with cellFRAP is intuitive and simple as the system is completely integrated into cellSens, the Olympus Life Science imaging platform. An automated calibration procedure makes it easy to perfectly align the scanner with the camera while taking into account the objectives and laser lines in use. Operating cellFRAP in the cellSens Graphical Experiment Manager offers intuitive system settings and experiment control, and it allows utmost operational flexibility and optimizes for the best speed performance. Thus, even complex acquisition workflows can be easily defined and reproduced, and any device operation can be automated.

Interactive Experiment Control
cellFRAP makes it possible to precisely adapt the bleaching regions to sample structures. An interactive definition of experimental conditions includes a free choice of bleaching/photomanipulation regions—variable in shape, including points and lines. Two different operation modes are available: Manipulation either based on predefined regions of interest or via interactive “Click & Bleach” mode. Signal intensity changes over time can also be observed online by displaying an intensity profile on demand.

Choose Your Method of Evaluating Data
In order to accurately evaluate your data, time markers for the start and end of the photomanipulation event are added to the recorded data. cellFRAP includes a qualitative FRAP analysis to classify your results. Here, recovery rates and mobile and immobile fractions can quickly be calculated from the recorded data including background and bleaching corrections. In addition, a Kymograph resembles a timeline visualization along a user-defined trajectory. An integrated export function allows for further evaluation of the data.

Flexibility According to Your Needs
Integrated within the Olympus IX3 open-source concept, the cellFRAP deck solution can easily be adapted to an existing imaging platform and combines with a wide range of accessories like ZDC, climate chambers, manual or motorized stages, manipulators, etc. This allows for the setup of photomanipulation experiments in combination with other high-end applications—for example, spinning-disk and TIRF microscopy. The system can be equipped with up to four lasers for photomanipulation, a prerequisite for further sophisticated applications such as distinct photoswitching or optogenetics.