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Confocal Laser-scanning Microscopy in Filamentous Fungi

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Advanced Microscopy in Mycology

Part of the book series: Fungal Biology ((FUNGBIO))

Abstract

Confocal microscopy is an advanced fluorescence microscopy technique that utilizes laser illumination, optical devices, pinholes, computers, and cameras/detectors to perform cell and subcellular visualization. Confocal means “having the same focus” in terms of microscopy; the imaged region captured by the detector will be the same (i.e., in focus) as the in-focus illuminated region in the specimen. Thus, the out-of-focus light emanating from above and below the focal plane of the specimen is not included in the final image and only a thin optical plane of the sample is in focus. This “optical sectioning” is the principal advantage of confocal microscopy and is achieved without physically sectioning the specimen. Confocal images have improved resolution and contrast compared to those obtained with a conventional wide-field epifluorescence microscopy. Such an advantage is achieved by the use of monochromatic lasers to illuminate and scan across a specimen, and pinholes to eliminate out-of-focus light. Confocal microscopy has been successfully applied to answer a wide variety of fungal biology questions including cell and cytoplasmic dynamics and structure, fungus–host interactions, and organelle structure and function using different reporters simultaneously. This chapter focuses primarily on confocal microscopy used to image fungal cells. As with any technique, in spite of its great advantages, the pros and cons of confocal microscopy must be considered when determining its potential applications for specific fungal samples.

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Authors and Affiliations

  1. Department of Microbiology, Center of Scientific Research and Higher Education of Ensenada, 3918 Carretera Ensenada-Tijuana, 22860, Ensenada, B. C., Mexico

    Dr. Rosa R. Mouriño-Pérez

  2. School of Life Sciences, Arizona State University, Tempe, AZ, USA

    Dr. Robert W. Roberson

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  1. Dr. Rosa R. Mouriño-Pérez
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Correspondence to Rosa R. Mouriño-Pérez .

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Editors and Affiliations

  1. University of Regina, Regina, Canada

    Tanya E. S. Dahms

  2. North American Applications and Labs, Carl Zeiss Microscopy, LLC, Thornwood, New York, USA

    Kirk J. Czymmek

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Mouriño-Pérez, R., Roberson, R. (2015). Confocal Laser-scanning Microscopy in Filamentous Fungi. In: Dahms, T., Czymmek, K. (eds) Advanced Microscopy in Mycology. Fungal Biology. Springer, Cham. https://doi.org/10.1007/978-3-319-22437-4_1

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