Here, we extend our previous studies on imaging yeast cells to image subcellular architecture in human melanoma cells and melanocytes at resolutions as high as ∼6 and ∼20 nm in the directions parallel and perpendicular, respectively, to the direction of ion-beam milling.
With the binary images, a yeast candidate mask image can be constructed by combining the GF (green fluorescence) and RF (red fluorescence) binary images.
When we observe the histograms of intensities in the image of yeast samples, they are easily modeled as a Gaussian distribution.
doi: 10.7554/eLife.00367 Image Individual yeast cells (yellow) evolve to form multicellular clumps (magenta and green) We typically think of single-celled microbes as being solitary creatures endowed with an innate self-sufficiency.
doi: 10.7554/eLife.02630 Image Hybrid yeast (right) produce fewer viable spores and smaller colonies than either parent species (left, middle) Cheaters who act selfishly to prosper at the expense of others are commonplace in the natural world, and genomes are no exception.
A DeltaVision fluorescence microscopy system (Applied Precision), which is based on an Olympus wide-field IX71 fluorescence microscope equipped with an oil-immersion objective lens (Plan Apo 60×; NA = 1.4; Olympus) and CoolSNAP HQ2 CCD camera (Photometrics), was used to image the yeast cells.
Images of yeast culture plates with GFP fluorescence were obtained using a LAS4000 lumino-image analyzer (Fujifilm) and were processed using Photoshop CS2 software (Adobe).
Gel images from yeast fingerprints were recorded under UV light in a digital photo-documenting apparatus (Bio-Rad Laboratories, HerCAles, CA, USA).
Electron microscopy reveals high-quality images of yeast spliceosomes at defined functional stages, indicating that they are well-suited for three-dimensional structure analyses.
To assess nuclear (spindle) movement, images of yeast cells expressing GFP-Tub1p were taken at 30 sec intervals for a total of 60 min, with an exposure time of 500 msec at each focal plane.
