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Shadow Technique Algorithm (STA) Sheds a New Light on Differential Interference Contrast (DIC) Microscopy

Dave Trinel, Pauline Vandame, Magalie Hervieu, Emilie Floquet, Marc Aumercier, Emanuele G Biondi, Jean-François Bodart and Corentin Spriet

Diversity of biological samples is still partially considered by conventional Differential Interference Contrast (DIC) microscopy approaches. Here we propose a new algorithm (developed as an ImageJ macro), the STA (Shadow Technique Algorithm), whose originality relies in the 3D/4D visualization of a large range of biological objects, from bacteria, vegetal tissues to living cells in culture. This new approach does not need extensive calculations, systems modifications or in-depth knowledge of acquisition optics. STA, providing 3D DIC reconstruction every hundredth of ms, can be applied to dissect various cellular phenomena. In addition, we propose different methods of graphic representations, which unable to enlighten the specificities of each category of questioning. Specifically we here addressed: i) tissue imaging, ii) cell cycle and cell death imaging iii) vesicle tracking.