<Prof. Don Lamb>

Prof. Don Lamb

Advanced Fluorescence Microscopy Techniques

Development of the microscope is probably the single most important physical innovation for the life sciences. In the last couple of decades, new optical microscopy have been developed including the development of fluorescence fluctuation spectroscopies and super-resolution methods that have a broad range of applications in the physical, chemical and biological sciences.

In this talk, I will give a basic overview of some of these advanced methods and highlight their application with examples. I will give a brief introduction to fluorescence fluctuation spectroscopy (FFS) and how we can extract quantitative information out of fluctuations. FFS can be used to investigate molecular mobility and interactions.

In the second part of this talk, I will introduce the different types of super-resolution spectroscopy, structure illumination spectroscopy, stochastic optical reconstruction microscopy (STORM) (or photoactivation localization spectroscopy, PALM), and stimulated emission depletion (STED) microscopy. The advantages, limitations and accuracies of the different methods will be discussed. We will also discuss super-resolution optical fluctuation imaging (SOFI), which utilizes fluorescence fluctuation spectroscopy to create super-resolution images.

In the last couple of minutes, time permitting, I will introduce a novel orbital tracking microscope that we have developed that can track single particles in real time with millisecond resolution and nanometer accuracy over centimeters and hours. We have used this microscope to track mitochondria transport in Zebra fish embryos.