Practical considerations of objective lenses for application in cell biology.

For nearly a century, examination of biological phenomena on a microscopic level has depended on carefully calibrated optical systems, with the objective lens being regarded as the critical determinant of image quality. In this modern day, a wide variety of high-quality objectives exist, many with highly specialized functions and all requiring at least a certain base level of knowledge on the part of the imager in order to realize their full potential. A good working knowledge of objective construction, proper use, specialized applications, and care goes a long way toward obtaining quality results. Presented here is a practical guide to choosing, using, and maintaining the proper objective for a given biological imaging application.

Structured illumination microscopy for superresolution.

The ability to image beyond the diffraction limit is the central tenet of the burgeoning field of superresolution fluorescence microscopy, also referred to as optical nanoscopy. The advent of superresolution has revolutionized biological fluorescence microscopy and the field at large. However, much of that excitement has been tempered by prohibitive imaging requirements. Achieving superresolution entails certain sacrifices, namely imaging speed, choice of fluorophore, ease of multicolor and three-dimensional imaging, and generally more complex instrumentation as compared to standard widefield imaging techniques. Several techniques utilizing structured illumination occupy an intriguing middle ground between the ease of use associated with traditional fluorescence microscopies and the unprecedented resolving power of modern superresolution methods, resulting in undeniably robust imaging techniques. Presented here is a review of the conceptual basis of structured illumination and its implementation, including its performance in comparison to other nanoscopies and the most recent developments in the field.

Sample preparation for single molecule localization microscopy.

Single molecule localization-based optical nanoscopy was introduced in 2006, surpassing traditional diffraction-limited resolutions by an order of magnitude. Seven years later, this superresolution technique is continuing to follow a trend of increasing popularity and pervasiveness, with the proof-of-concept work long finished and commercial implementations now available. However one important aspect that tends to become lost in translation is the importance of proper sample preparation, with very few resources addressing the considerations that must be made when preparing samples for imaging with single molecule level sensitivity. Presented here is a an in-depth analysis of all aspects of sample preparation for single molecule superresolution, including both live and fixed cell preparation, choice of fluorophore, fixation and staining techniques, and imaging buffer considerations.

Phenylbutyrates as potent, orally bioavailable vitronectin receptor (integrin alphavbeta3) antagonists.

In our continuing efforts to identify small molecule vitronectin receptor antagonists, we have discovered a series of phenylbutyrate derivatives, exemplified by 16, which have good potency and excellent oral bioavailability (approximately 100% in rats). This new series is derived conceptually from opening of the seven-membered ring of SB-265123.