This course provides an introduction to the principles that govern the acquisition of 3D images with optical microscopes. Specifically, it provides attendees with practical knowledge to understand the limitations of conventional microscopes when imaging 3D samples, as well as the principles of different emerging microscopy techniques with optical-sectioning capacity The course will include three parts. In the first part, we describe the fundamentals of 2D imaging processes in conventional microscopes, and why they are not well adapted for imaging 3D samples. In the second part, we will focus on different optical-sectioning microscopy techniques, such as confocal, 4Pi, multi-photon, and structured illumination microscopy. In the third part, we will focus on emerging approaches, including one-shot 3D microscopes, digital holographic microscopes, etc. The attendee will benefit from a concise and realistic overview of microscopy procedures, which may help them to select the adequate microscope for various applications. The course will provide discussions of optical hardware and various practical applications of 3D optical microscopy. Also, discussions and examples will be presented on the benefits of 3D optical microscopy over conventional 2D optical microscopy.

The course will review the following fundamentals which are necessary to understand, design, and analyze 3D imaging and display systems. The course helps the students to understand how 3D imaging works, what are the fundamentals, how to use optics to implement 3D displays, how to improve performance in 3D imaging systems and/or product lines and new product development programs, how to increase optical performance, or simply find new solutions to existing technological problems. Fundaments of Geometrical Optics: propagation of rays in transparent materials, refraction of rays in plane and spherical diopters, image formation with lenses, combination of lenses, aperture and field limitation, law of lenses: image position and magnification, examples and applications. Wave theory of image formation: the plane wave and the spherical waves, the wavefield as linear superposition of spherical waves, propagation of wavefields though converging lenses, waves through telecentric optical systems, image formation analyzed in terms of wave optics: the concepts of PSF, spatial resolution, OTF and frequency cut-off, light diffracted through periodic screens, examples and applications. Wave and ray theory of 3D optical capture and display systems such as plenoptic systems: capture of lightfield with an array of digital cameras: the synthetic aperture method, capture of lightfield with a plenoptic camera working in the 1.0 mode, capture of lightfield with a plenoptic camera working in the 2.0 mode, algorithms for the calculation of views and for the reconstruction in depth, examples and applications, implementation of synthetic-aperture setup, and implementation of a plenoptic camera.