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Article Abstract
This review is intended as a guideline for beginners in confocal laser scanning microscopy. It combines basic theoretical concepts, such as fluorescence principles, resolution limits, and imaging parameters with practical guidance on sample preparation, staining strategies, and data acquisition using confocal microscopy. The aim is to combine technical and methodological aspects in order to provide a comprehensive and accessible introduction. The main application is in microbiology, with a focus on biofilms and bio-aggregates; although other researchers in biology may benefit from this survey. In this primer, we compiled 25 years of experience with confocal microscopy, four generations of instruments, diverse national and international projects, and many different samples of PhD students, PostDocs, and senior scientists from various countries.
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