BioFilmAnalyzer

BioFilmAnalyzer quantifies sub-population fractions in biofilms and adherent tissue samples from fluorescent microscopic and confocal laser scanning microscopy (CLSM) images to enable analysis of cells that cannot be dissociated for flow cytometry.

Key Features:

• Fluorescent and CLSM image analysis: Processes fluorescent microscopic images and confocal laser scanning microscopy (CLSM) images to evaluate cell sub-population fractions.
• Two-step algorithm: Implements a two-step algorithm where single-cell selection uses objective cell size distribution analysis followed by statistical quantification of sub-population fractions.
• Batch sequential analysis: Performs sequential analyses on homogeneous series of similar microscopic images without further intervention.
• Validation against manual counting and flow cytometry: Validated with experimental measurements on bacterial and eukaryotic cells showing agreement with manual microphotograph counting and flow cytometry, including live/dead staining assessments.

Scientific Applications:

• Biofilm and adherent cell analysis: Quantifying sub-populations in bacterial biofilms and adherent eukaryotic cell lines from microphotographs.
• Live/dead staining quantification: Measuring live/dead cell fractions within biofilm-embedded and surface-adherent cells.
• Comparative image-based validation: Providing image-derived estimates comparable to flow cytometry and manual counts for studies where cell dissociation is impractical.

Methodology:

Computational analysis of fluorescent and CLSM microphotographs using a two-step algorithm: objective cell size distribution–based single-cell selection followed by statistical quantification of sub-population fractions.