Image analysis algorithms for computer vision to instruct microscope operations in real-time (crops, change in frame rate, photobleaching, z-stack imaging)
Moreover, Image analysis is not a magic tool that outputs quantitative information from any image. It is intimately linked to the imaging process. My experience with microscopy will help you optimise the planning of your imaging experiment to maximise the quality of the image analysis quantitative output.
Centroid tracking of an endocytic coat marker (Sla1) tagged with EGFP and imaged as a diffraction limited spot in wide-field microscopy. Sla1 dynamic acts as a proxy for the endocytic invagination growth (Picco, et al., 2015). The CLEM image on the right shows a mature endocytic invagination for comparison (Kukulski, et al., 2012). Scale bar is 100 nm and applies to both images.Cell tracking of a Fonticula Alba Lobose Amoeba imaged with phase contrast microscopy
(Toret, et al., 2022).Stoichiometry of the ERMES complex computed by ratiometric comparison of fluorescence intensity of diffraction-limited puncta of EGFP-tagged Cse4 (grey), Mmm1 (orange), Mdm12 (purple) and Mdm34 (green), represented as dot plots as well as half-violin plots in (C). Left column indicates number of analyzed puncta. Right column indicates numbers of EGFP molecules/punctum derived from fluorescence intensities using Cse4-EGFP as reference (median values with MAD are given).
(Wozny, et al., BiorXiv, 2022, Wozny, et al., Nature, 2023). For the complete legend see the Stoichiometry of the ERMES complex.
