TRACMIT

TRACMIT performs high-throughput tracking and analysis of individual human cells on micropatterns to identify mitotic events and quantify mitotic spindle orientation during metaphase from time-lapse fluorescence microscopy (e.g., mCherry::histone 2B).

Key Features:

• Fiji/ImageJ integration: Implemented as a series of macros running within the Fiji/ImageJ platform to leverage established image-processing functions.
• High-throughput analysis: Enables parallel analysis of large numbers of cells suitable for screening-scale time-lapse datasets.
• Micropattern filtering: Identifies micropatterns containing a single viable cell and excludes dying or compromised cells for focused analysis.
• Automated spindle orientation assessment: Determines mitotic spindle orientation during metaphase in human cells expressing mCherry::histone 2B by tracking back from anaphase figures in time-lapse recordings to assess spindle angle relative to the micropattern.
• Manual validation support: Provides the ability to perform manual validation of selected cell traces to confirm automated selections and measurements.
• Micropattern shape flexibility: Supports analysis of cells plated on various micropattern shapes for adaptable experimental designs.
• Data efficiency: Processes raw data directly without saving intermediate images to reduce storage and I/O overhead.

Scientific Applications:

• Spindle positioning studies: Quantification of mitotic spindle orientation during metaphase in human cells expressing mCherry::histone 2B.
• Cell division and mitosis research: Analysis of mitotic events and phenotypes at single-cell resolution across large datasets.
• Cytoskeletal dynamics: Investigation of cytoskeleton-related phenotypes linked to spindle orientation and mitotic progression.
• High-throughput screening: Large-scale screening of dynamic cellular phenotypes using time-lapse fluorescence microscopy on micropatterned substrates.

Methodology:

Implemented as Fiji/ImageJ macros that filter micropatterns to single viable cells, track back from anaphase figures in time-lapse fluorescence recordings (mCherry::histone 2B) to measure metaphase spindle angle relative to the micropattern, and process raw data without saving intermediate images.