MAG Biosystems™ is pleased to introduce FRAP-3D™, the world’s first widefield imaging system designed specifically to study the intracellular dynamics of proteins and other macromolecular complexes via high-speed, multidimensional FRAP (fluorescence recovery after photobleaching) and iFRAP (inverse FRAP) experiments. Photoactivation and photoconversion studies with fluorescent proteins such as PA-GFP, EOS, KFP, Kaede, and Dronpa can also be performed.
FRAP-3D includes a galvanometer-based FRAP head, an innovative laser launch module with integrated I/O circuitry to control all aspects of the system, the world’s first and only quantitatively stable 16-bit EMCCD camera (the QuantEM™), acquisition and analysis software, and a configured workstation. The system’s FRAP head mounts easily to many popular inverted microscopes through the epi-illumination port.
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Use FRAP-3D to study life's dynamic details!
The best method for measuring global protein diffusion in cells is FRAP. In this technique, the molecule of interest is fluorescently labeled and a focused laser is used to rapidly and acutely inactivate the fluorescence by photobleaching a small (ideally diffraction-limited) subregion of the cell. Mobile molecules then repopulate the bleached area over time. The rate of recovery provides an accurate measure of the molecular mobility within the cell.
The FRAP-3D system has been designed specifically for a number of live-cell imaging applications, including:
FRAP and iFRAP studies
Acceptor photobleaching
Photoactivation studies with PA-GFP, Dronpa, and KFP
Photoconversion studies with EOS, Kaede, and Dendra
Capture data quickly and easily!
mEmerald recovery in U2 cells. Data collected in Michael Davidson’s lab at Florida State University.
FRAP-3D is engineered to be the fastest FRAP imaging system available:
Burst mode captures the first post-bleach image less than 25 msec from the end of the last bleach pulse, enabling the study of rapidly diffusing macromolecules
Custom optics enable simultaneous FRAP and widefield illumination
Absence of moving parts in the optical path allows the system to switch quickly between bleach and recovery phases
High-speed EMCCD acquires 30 fps at full resolution for monitoring rapid protein dynamics
Detect even the weakest fluorescence signal!
The FRAP-3D system utilizes the world’s first and only quantitatively stable 16-bit EMCCD camera, the Photometrics QuantEM. This electron-multiplying, 512 x 512-pixel sensor delivers greater than 90% peak quantum efficiency with extremely low read noise to ensure reliable detection of ultra-low-light fluorescence signal.
Never realign or reconfigure imaging hardware again!
The FRAP-3D system’s galvanometer-based FRAP head can be integrated easily with many popular inverted microscopes. Built-in custom optics are used to combine laser illumination with widefield illumination in a single beampath, thus permitting simultaneous FRAP and widefield imaging.This unique high-precision design also lets researchers perform standalone full-widefield imaging without changing optics. The FRAP-3D system never requires alignment by the user.
Control your whole experiment with easy-to-use software!
FRAP-3D software utilizes an intuitive, easy-to-use graphical user interface (GUI) that gives researchers complete control of all acquisition-related system parameters.
Live FRAP w/ Instant Recall™ Perform real-time FRAP experiments to assess FRAP efficiency and recovery times. Select Save to enable Instant Recall. This exclusive feature lets researchers acquire a complete FRAP experiment via a single click of the mouse while viewing their sample in real-time.
Analyze your data automatically!
A single click within the FRAP-3D menu bar brings up the FRAP-3D Analyzer window, where all data analysis is performed automatically. More advanced analysis tools are also available to allow researchers to tailor the analysis to their experimental needs.
Acquire quantitative 3D data over time!
The FRAP-3D system offers the ability to acquire quantitative pre- and post-bleach data in 3D over time. Researchers can define bleach regions specific to defined z-planes through the FRAP-3D software, which supports any piezo focus device via analog voltage (0–10 V). Deconvolution analyses can be performed when high spatial resolution is required.
Utilize up to four lasers!
The FRAP-3D system features an advanced, compact laser launch that supports up to four diode or DPSS lasers. Custom dichroics, specifically designed for the laser combinations selected, combine the individual beams. Precise control over each laser’s illumination intensity is provided via a four-channel AOTF with a separate blanking channel.
Data collected in Michael Davidson’s lab at Florida State University.
