OBCOL

OBCOL performs organelle-level segmentation and quantitative colocalisation analysis in 2D, 3D and 4D multichannel fluorescence microscopy images to quantify protein distribution and overlap within individual organelles.

Key Features:

• Automatic Segmentation: Automates segmentation of individual punctuate organelles within 3D multichannel images.
• Quantitative Colocalisation Analysis: Calculates a variety of colocalisation statistics for segmented objects, including positional data, degree of overlap between fluorescent channels, and the number of component objects per organelle.
• Multidimensional Imaging Support: Supports analysis of 2D, 3D, and 4D imaging modalities.
• Organelle-level Analysis: Focuses analysis on individual organelles rather than entire images to resolve subcellular distribution and interactions.

Scientific Applications:

• Fluorescent marker validation: Validated using 3D microscopy imaging of Dextran and EGF to assess marker distribution.
• Recycling endosome analysis: Applied to quantify colocalisation of VAMP3, Rab11, and transferrin on recycling endosomes in mammalian cells, revealing diversity of endosome labelling and overlap within individual recycling endosomes.

Methodology:

Segmentation of organelles followed by detailed colocalisation analysis on each segmented object.