Microscopy: Imaging Systems and Techniques

This lecture introduces the fundamentals of microscopy and imaging systems, explaining the need for microscopes to magnify objects for better visibility. It covers topics such as the functioning of lenses, the concept of near point, and the use of magnifying glasses. The lecture delves into the principles of geometrical optics, discussing convex lenses, focal distances, and the creation of real and virtual images. It also explores compound microscopes, infinity microscopes, and microscope objectives, emphasizing the importance of quality objectives for image clarity. Additionally, it touches on darkfield and phase contrast microscopy, fluorescence microscopy, and super resolution techniques like structured illumination microscopy and STED. The lecture concludes by highlighting the significance of super resolution microscopy in overcoming the diffraction barrier.

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The course deals with the concept of measuring in different domains, particularly in the electrical, optical, and microscale domains. The course will end with a perspective on quantum measurements, wh

Microscopy is the technical field of using microscopes to view objects and areas of objects that cannot be seen with the naked eye (objects that are not within the resolution range of the normal eye). There are three well-known branches of microscopy: optical, electron, and scanning probe microscopy, along with the emerging field of X-ray microscopy. Optical microscopy and electron microscopy involve the diffraction, reflection, or refraction of electromagnetic radiation/electron beams interacting with the specimen, and the collection of the scattered radiation or another signal in order to create an image.

The optical microscope, also referred to as a light microscope, is a type of microscope that commonly uses visible light and a system of lenses to generate magnified images of small objects. Optical microscopes are the oldest design of microscope and were possibly invented in their present compound form in the 17th century. Basic optical microscopes can be very simple, although many complex designs aim to improve resolution and sample contrast. The object is placed on a stage and may be directly viewed through one or two eyepieces on the microscope.

Confocal microscopy, most frequently confocal laser scanning microscopy (CLSM) or laser scanning confocal microscopy (LSCM), is an optical imaging technique for increasing optical resolution and contrast of a micrograph by means of using a spatial pinhole to block out-of-focus light in image formation. Capturing multiple two-dimensional images at different depths in a sample enables the reconstruction of three-dimensional structures (a process known as optical sectioning) within an object.

NOTOC Phase-contrast microscopy (PCM) is an optical microscopy technique that converts phase shifts in light passing through a transparent specimen to brightness changes in the image. Phase shifts themselves are invisible, but become visible when shown as brightness variations. When light waves travel through a medium other than a vacuum, interaction with the medium causes the wave amplitude and phase to change in a manner dependent on properties of the medium.

A microscope () is a laboratory instrument used to examine objects that are too small to be seen by the naked eye. Microscopy is the science of investigating small objects and structures using a microscope. Microscopic means being invisible to the eye unless aided by a microscope. There are many types of microscopes, and they may be grouped in different ways.

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