TIKAL

TIKAL performs 4-D image processing of laser scanning and wide-field microscopy data to quantitatively track nuclear bodies and ribonucleoprotein particles (RNPs) and correlate their dynamics with chromatin organization.

Key Features:

• 4-D image processing: Processes time-resolved 3-D image stacks from laser scanning and wide-field microscopes for dynamic analysis.
• Registration: Corrects global cellular movements and localized deformations to preserve spatial accuracy during time-lapse sequences.
• 2-D and 3-D tracking: Performs single-particle tracking in two and three dimensions to extract particle trajectories over time.
• Concurrent chromatin analysis: Acquires chromatin density data alongside particle positions to enable direct correlation of particle dynamics with chromatin reorganization.
• Kinetic classification: Identifies four modes of particle movement: confined obstructed, normal diffusion, directed motion, and a mode influenced by local chromatin changes.
• Single-particle mobility analysis: Quantifies mobility parameters of particles such as GFP-NLS-vimentin nuclear bodies and microinjected polystyrene beads.

Scientific Applications:

• Nuclear particle dynamics: Quantitative analysis of GFP-NLS-vimentin nuclear bodies and microinjected polystyrene beads in living cells.
• Cells with complex motion: Tracking and analysis in cells exhibiting global movements, local deformations, and nuclei undergoing shape changes.
• Chromatin–particle correlation: Studying how local chromatin organization and chromatin compaction (e.g., 0.6 M sorbitol dehydration) affect particle mobility.
• Perturbation studies: Assessing effects of ATP levels, actin and microtubule integrity, and nuclear hydration state on particle kinetic behavior and differences between nuclear and cytoplasmic environments.

Methodology:

Image registration correcting global and local deformations; 2-D and 3-D single-particle tracking; automatic 3-D particle movement analysis concurrent with chromatin density acquisition; and kinetic analysis classifying movements into four modes (confined obstructed, normal diffusion, directed motion, chromatin-influenced).