Analytical and Quantitative Light Microscopy

Course Description

A comprehensive and intensive course in light microscopy for researchers in biology, medicine, and material sciences. This course provides a systematic and in-depth examination of the theory of image formation and application of video and digital methods for exploring subtle interactions between light and the specimen. This course emphasizes the quantitative issues that are critical to the proper interpretation of images obtained with modern wide-field and confocal microscopes. This course is limited to 32 students.

Laboratory exercises, demonstrations, and discussions include: (1) geometrical and physical optics of microscope image formation including Abbe’s theory of the microscope and Fourier optics; (2) interaction of light and matter; (3) phase contrast polarization and interference microscopy for the nondestructive analysis of molecular and fine-structural organization in living cells; (4) fluorescence microscopy, quantification of fluorescence, and GFP; (5) principles and application of digital video imaging, recording, analysis, and display; (6) digital image processing and quantitative digital image deconvolution; (7) ratiometric measurement of intracellular ion concentrations; (8) confocal microscopy; and (9) new advances in light microscopy such as FRET, FLIM, TIRF, and patterned illumination.

The program is designed primarily for: (1) university faculty, professional researchers, postdoctoral fellows, and advanced graduate students in the life sciences who wish to expand their experience in microscopy and to understand the quantitative issues associated with analysis of data obtained with optical microscopes; (2) individuals well-grounded in the physical sciences, who wish to exploit microscopy techniques for analyzing dynamic fine-structural and chemical changes; and (3) industrial scientists and engineers interested in advancing the design of equipment and techniques involving video and digital microscopy.

Lectures are followed by small group laboratory sessions and demonstrations. As a result, students will have opportunities for extensive hands-on experience with state-of-the-art optical, electronic, and digital imaging equipment guided by an experienced staff from universities and industry.