Label-free dynamic segmentation and morphological analysis of subcellular optical scatterers

Area of Science:

• Cell biology
• Biophysics
• Optical imaging

Background:

• Label-free imaging avoids artifacts from fluorescent proteins or dyes.
• Studying living cells requires minimal sample preparation and preserves cellular integrity.

Purpose of the Study:

• To develop a label-free imaging technique for quantifying cellular injury.
• To analyze dynamic changes in subcellular structures during induced cell injury.

Main Methods:

• Combined label-free optical scatter imaging with digital segmentation and processing.
• Created dynamic subcellular masks to highlight scattering objects.
• Quantified organelle morphology and redistribution during calcium overload-induced cell injury.

Main Results:

• Object aspect ratio and orientedness decreased in calcium-overloaded cells.
• Textural features of scatterer distribution changed significantly (>30%) in injured cells.
• Results correlated with fluorescence-based observations of mitochondrial fission and rounding.

Conclusions:

• Label-free morphological signatures can effectively indicate cellular injury.
• Dynamic analysis of subcellular scatterers offers a method to quantify cellular state changes.
• This technique provides a sensitive, artifact-free approach to cell injury assessment.