FIND YOUR WAY THROUGH THE JUNGLE OF COMPETING FLUORESCENCE IMAGING TECHNOLOGIES
This seminar is intended to provide guidelines for finding the right microscope for your application through the following topics of discussion:
Sample preparation - Tissue clearing methods
High end fluorescent technologies and the latest developments and trends
Strengths and limitations of various systems available – which compromises do I face – speed vs resolution vs sensitivity
Data in context - Correlative approaches – combining data from different instruments from Light to electron microscopy.
Cell culture has become an everyday useful tool to study the effects of compounds on the normal physiology of cells. With recent technology advances, there is a drive to increase the physiological relevance of data by studying more complex cell models eg. Spheroids, organoids, whole brain, whole animal embryo models like drosophila and zebrafish. As all scientist who are venturing into these exciting new realms have very early on realized, the biggest challenge in working with these advanced cell models is to find the right tool to see deeper and get more data out.
Fluorescence microscopy has become the most popular and widely used imaging modality over the years. This is particularly due to the outstanding specificity fluorescent markers convey to a protein of interest. Among the most prominent of these tags are genetically encoded fluorescent proteins (FPs) or antibodies conjugated to organic dyes. With better fluorescent tags emerging many wide-field and confocal technologies have evolved to cope with the ever-increasing demands for increasing acquisition speed, for obtaining better resolution, for being higher in sensitivity and what is more, to reduce light induced phototoxicity at the same time. Alas, like in real life, a microscope cannot serve all masters. For example, if speed must be increased to resolve a fast-dynamic process, sensitivity will suffer as exposure times have to be reduced. If one wants to counteract sensitivity loss by increasing the light dosage, increasing phototoxicity can become an issue. Thus, all parameters cannot be optimized simultaneously, and no system exists that will excel in all of them.
Consequently, different specimens and applications will pose different challenges on the performance of a fluorescence microscope. The choice of the instrument that serves one’s needs best is not only essential for being successful for the designed experiments but also crucial for being cost and time effective. The plethora of methods and specialized systems, however, can render this task rather tedious and difficult.
ZEISS WORKSHOP PRESENTER
Dr Klaus Weisshart
CARL ZEISS MICROSCOPY GmbH | GERMANY
Dr Klaus graduated in Biology at the University of Constance and obtained his PhD from the German Cancer Research Center in Heidelberg. He continued his work on DNA replication during research stays at Harvard Medical School Boston and the Ludwig-Maximilian-University in Munich. In the year 2000 he joined Carl Zeiss where he supports the product lines for fluctuation analysis, super-resolution and light sheet microscopy.