Reflection and rotation for three dimensional microscopy of live cells

Abstract

Confocal scanning laser microscopy and multiphoton microscopy provide 3D data from biological specimens, but with limited z-axis precision. Multiple microscale mirrors can be used to obtain more accurate 3D data on living cells while using classical widefield microscopy. Etched silicon wells coated with aluminum were used to obtain 3D images of budding yeast cells, with information along the z-axis provided by reflections from the angled sides of the well. To supplement measurements obtained with this method, work was also conducted on a system to allow for rotation of a cell attached to a pulled glass tip. Images of a yeast cell obtained from either method can be fit to a simple 3D surface due to the cell's roughly spheroidal shape, which should provide accurate measurements of the volume of an individual budding yeast cell as it progresses through the cell cycle.