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Automated detection of fluorescent cells in in‐resin fluorescence sections for integrated light and electron microscopy
Author(s) -
DELPIANO J.,
PIZARRO L.,
PEDDIE C.J.,
JONES M.L.,
GRIFFIN L.D.,
COLLINSON L.M.
Publication year - 2018
Publication title -
journal of microscopy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.569
H-Index - 111
eISSN - 1365-2818
pISSN - 0022-2720
DOI - 10.1111/jmi.12700
Subject(s) - fluorescence , fluorescence microscope , tracking (education) , computer science , microscopy , data acquisition , materials science , process (computing) , computer vision , microscope , resolution (logic) , automation , fluorescence lifetime imaging microscopy , electron microscope , artificial intelligence , biomedical engineering , optics , physics , engineering , mechanical engineering , psychology , pedagogy , operating system
Summary Integrated array tomography combines fluorescence and electron imaging of ultrathin sections in one microscope, and enables accurate high‐resolution correlation of fluorescent proteins to cell organelles and membranes. Large numbers of serial sections can be imaged sequentially to produce aligned volumes from both imaging modalities, thus producing enormous amounts of data that must be handled and processed using novel techniques. Here, we present a scheme for automated detection of fluorescent cells within thin resin sections, which could then be used to drive automated electron image acquisition from target regions via ‘smart tracking’. The aim of this work is to aid in optimization of the data acquisition process through automation, freeing the operator to work on other tasks and speeding up the process, while reducing data rates by only acquiring images from regions of interest. This new method is shown to be robust against noise and able to deal with regions of low fluorescence.