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Eva-Maria Weiss, Aneta Petruskova, Anna Fejtova (Department Psychiatry and Psychotherapy, University Hospital Erlangen,Erlangen, Germany;Charles University, Third Faculty of Medicine, Prague, CZ;National Institute of Mental Health, Klecany, CZ)
Synaptic transmission and synaptic plasticity areessential processes for learning and memory and determined by post- and pre-synaptic interplay.Appropriate antibodies allowlabeling pre- and postsynaptic puncta on fluorescence microscopy images. However, especially for methods that rely on detection of centroids or on creation of a binary image to localize synapsesand therefore require an overall intensity threshold, detectionis challenging due to low signal to noise ratio and heterogeneous brightness of synaptic puncta.We developed a Matlab-based synaptic detection tool based on an edge detection filter followed by separation of regions by a marker controlled watershed segmentation. Edge filter based pre-segmentation is free ofan overall intensity cut-off threshold andenablesinclusion of darkpuncta since gradient values are crucial. After flood filling and dilation,a marker-controlled watershed segmentation algorithm separates connected synaptic puncta and is robust against oversegmentation. Moreover, the tool provides calculations for relevant parameters for pre- and post-synapses properties, overlapping and co-localization.Here we provide a fast and unsupervised method for 2D flu