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Trends in imaging fungal pathogens for cell biological studies of plant disease

Trends in imaging fungal pathogens for cell biological studies of plant disease

Phytopathology 91(6 Supplement): S162

Advances in instrumentation and technique have accelerated the evolution of imaging in cell biology. This presentation will include examples of our recent work, and will cite examples from other laboratories, highlighting several new imaging applications in studies of fungal plant pathogens. The use of cryo-based preparative procedures remains the standard for fixed specimens at the tissue and cellular levels, for both light and electron microscope studies. For example, we have used immunocytochemistry to map the microtubule cytoskeleton in whole, intact, cryo-fixed hyphae, documented in three dimensions using digital imaging and confocal laser scanning microscopy. CryoSEM, as well as TEM after high-pressure freezing, continue to be important tools for investigations of host-pathogen ultrastructure. CLSM brings another dimension to the study of pathogenesis, in allowing for 3-D documentation of spatial relationships throughout the disease process. We demonstrate the utility of this approach for imaging infection hyphae in (a) fixed, cleared tissues, and (b) living or fixed tissues invaded by fungal transformants expressing spectral variants of green fluorescent protein. We anticipate an ever-increasing role for laser imaging, including multiphoton microscopy, and the use of living specimens to achieve maximum spatial and temporal resolution in studies of fungi.

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