Quantitative analysis of yeast internal architecture using soft X-ray tomography.

We used soft X-ray tomography (SXT)--a high-resolution, quantitative imaging technique--to measure cell size and organelle volumes in yeasts. Cell size is a key factor in initiating cell division in yeasts, whereas the number and volume of the organelles have a profound impact on the function and viability of a cell. Consequently, determining these cell parameters is fundamentally important in understanding yeast biology.

Live-cell imaging of microtubule dynamics in hyphae of Neurospora crassa.

Due to the large number of microtubules in the wild-type strain, total internal reflection fluorescence (TIRF) microscopy was used to study cortical microtubule dynamics in leading hyphae of Neurospora crassa expressing beta-tubulin-GFP. Detection of plus-end dynamics of individual microtubule was much improved with this approach compared to the other commonly used methods such as confocal and widefield fluorescence microscopy. In order to address the roles of motor proteins in microtubule dynamics, microtubule-motor mutant strains, deltankin and ro-1 were examined.

Soft X-ray tomography of phenotypic switching and the cellular response to antifungal peptoids in Candida albicans.

The opportunistic pathogen Candida albicans can undergo phenotypic switching between a benign, unicellular phenotype and an invasive, multicellular form that causes candidiasis. Increasingly, strains of Candida are becoming resistant to antifungal drugs, making the treatment of candidiasis difficult, especially in immunocompromised or critically ill patients. Consequently, there is a pressing need to develop new drugs that circumvent fungal drug-resistance mechanisms.

Soft X-ray tomography and cryogenic light microscopy: the cool combination in cellular imaging.

Soft X-ray tomography (SXT) is ideally suited to imaging sub-cellular architecture and organization, particularly in eukaryotic cells. SXT is similar in concept to the well-established medical diagnostic technique computed axial tomography (CAT), except SXT is capable of imaging with a spatial resolution of 50nm, or better. In SXT, cells are imaged using photons from a region of the spectrum known as the 'water window'.

Microtubule dynamics and the role of molecular motors in Neurospora crassa.

Live-cell imaging methods were used to study microtubule dynamics in the apical regions of leading hyphae and germ tubes of Neurospora crassa expressing beta-tubulin-GFP. Microtubule polymerization rates in hyphae of N. crassa were much faster than those previously reported in any other eukaryotic organism.

Application of electron tomography to fungal ultrastructure studies.

Access to structural information at the nanoscale enables fundamental insights into many complex biological systems. The development of the transmission electron microscope (TEM) has vastly increased our understanding of multiple biological systems. However, when attempting to visualize and understand the organizational and functional complexities that are typical of cells and tissues, the standard 2-D analyses that TEM affords often fall short.

In vivo effects of the fungicide ethaboxam on microtubule integrity in Phytophthora infestans.

The effects of ethaboxam (LGC-30473), a novel thiazole carboxamide, on microtubule organization was investigated in Phytophthora infestans (Mont) de Bary (Oomycota) at the light microscope level using standard immunofluorescence and laser scanning confocal microscopy. Hyphal cells were exposed to various concentrations ranging from 0.01 to 1.