KiT

KiT tracks kinetochores in live-cell fluorescence microscopy movies to provide quantitative localization and motion analysis for studying chromosome dynamics during mitosis.

Key Features:

• Versatile Imaging Support: Supports 2D, 3D, and multi-color movies for different imaging modalities.
• Quantitative Fluorescence Analysis: Performs fluorescence intensity quantification to assess protein dynamics and interactions.
• Integrated Deconvolution: Incorporates deconvolution to enhance image clarity and resolution for more accurate localization.
• Parallel Execution: Supports parallel execution to accelerate processing of large datasets.
• Multiple Localization Algorithms: Implements a range of particle/kinetochore localization algorithms to accommodate diverse experimental conditions.
• Regularization and Diagnostic Tools: Includes a regularization term in the objective function to encode biological constraints (e.g., known kinetochore counts) and provides diagnostic tools for verifying tracking accuracy.

Scientific Applications:

• Mitotic chromosome dynamics: Quantitatively analyzes kinetochore behavior from prometaphase through anaphase to study chromosome congression and segregation.
• High-resolution time-lapse imaging with LLSM: Applied to lattice light-sheet microscopy (LLSM) datasets to enable extended, high-resolution imaging with reduced phototoxicity for dynamic studies.

Methodology:

Automated quantitative tracking using particle/kinetochore localization algorithms, deconvolution, fluorescence intensity quantification, parallel execution, an objective function with a regularization term for biological constraints, and diagnostic tools for tracking verification.