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This module provides easy-to-use tools for both on-line and post-capture analysis and exploration of ratiometric data. This includes support for fura-2, indo-1 and BCECF imaging, fluorescence resonance energy transfer (FRET) experiments, and densitometry measurements. The module adds real-time graphing of ratios and intensities, background subtraction, and tracking of multiple objects. It also incorporates post-capture ratio generation, a ratio calibration guide and both direct sensitized FRET and acceptor photobleaching FRET capture protocols and transfer computations.
Ratio/Densitometry Features
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Real-time ratio computation during image acquisition
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Real-time graphing of ratios individual wavelengths during acquisition
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Real-time tracking of up to 20 ROIs
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Simultaneous imaging of two ratio indicators
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Calibration guide for computing nM ion concentration
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Analytical graphing tools
FRET Analysis
The module was developed in collaboration with a number of leading FRET biologists. The module contains analysis tools for both sensitized emission FRET and acceptor photobleaching FRET:
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Channel bleed-through calculation
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Background Subtraction
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Corrected FRET image generation (Herman Equation)
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Single pixel to whole object energy transfer measurement
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Support for 2D, 3D, and 4D data
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Real-time FRET analysis via support for dual camera capture and image splitting devices
Specifications
Ratio Capture and Analysis
• Real-time ratio region tracking and graphing
• Regions to mask transfer
• Supports simultaneous dual ratio capture (e.g. FURA-2 and BCECF)
• FURA-2 calibration guide
• Immediate display of automated ratio values or ion concentration
• Analytical graphing tools
• Multiple camera
Acceptor Photobleach FRET Capture and Analysis
• Automatic bleach capture script
• Automatic FRET computation
• Gated pseudocolor display
• Track FRET by object or pixel
• Background subtraction
Sensitized Emission FRET Capture and Analysis
• Live corrected FRET image generation
• Background subtraction
• Channel bleedthrough calculation
• FRET calibration guide
• Gated pseudocolor display
• Track FRET by object or pixel
Densitometry Capture and Analysis
• Interactive calibration guide
• Multiple curve fits
• Calibrations can be saved and applied to multiple images
