Microscopy: methods of visualisation in micro- and nano-scale

The continuous development of various approaches to the visualization of micro- and nanoscale objects is one of the most powerful drivers for the advancement of learning and scientific progress.  The fields like cell biology, microbiology, virology, biomedical studies, chemistry, material sciences, microelectronics, and nanotechnology benefit the most from these improvements. The ruling class of the instruments that provides researchers the data considering the structure of the systems that are too small to be discerned by the unaided eye are the microscopes of various types. The key feature of the widely speaking optical, encompassing both light and electron microscopy, is that the scientist employing the microscope obtains the data in a most comprehensive form - as an image. 

Within “Microscopy: Methods of Micro- and Nanoscale Visualization” course the students get acquainted with the physical principles underlying the functioning, possibilities and fundamental limitations of the broad spectrum of modern research microscopes.   The program includes wide-field transmitted light microscopes, wide-field, and laser-scanning fluorescent microscopes, optical super-resolution instruments of various designs, and also transmission and scanning electron microscopes. The program is primarily focused but not limited to the applications of microscopy to the studies of biological samples. Also, the course deals with the basic principles of acquiring of the microscopic images using modern digital light detectors and the ways of acquisition optimization for further extraction of quantitive data from “just the picture.” Also included are the application of deconvolution and approaches to the creation of three-dimensional models of the samples. 

The program integrates laboratory practice. During this stage, the students obtain practical skills required for the employment of the microscopes. This part includes the following topics: basic principles of manual microscope operation, initial adjustments, setting up Kohler illumination, application of the phase contrast and DIC, good habits, and safety requirements for the successful operation of the instruments. Also, students are introduced to the fluorescent techniques, digital imaging with the microscope and image calibration. The practical tasks also include the studies of the biological samples using a laser scanning confocal microscope. This part comprises the 3D imaging and dynamic FRAP experiments in a model system. The practical part also includes the experiments with simple methods of sample preparation for transmission electron microscopy and work with the microscope.   

The goal of the program is to introduce the students to the essential principles and necessary skills for studies micro- and nanoscale objects using modern microscopes.

The practical aims

1. Explain to the students the essential physical principles of the microscopes’ functioning, possibilities, and limitation of various types of instruments and the sample requirements demanded by those instruments.
2. Familiarize the students with the sample preparation techniques, so that they can make an informed choice and select the adequate approach if their research interest would require microscopy. 
3. Teach the students the most important practical aspects of the microscopes’ operation in the laboratory environment.
4. Provide students with the «keys» for the interpretation of microscopic images.

The position of the course within the program of the graduate 

The course “Microscopy: Methods of Micro- and Nanoscale Visualization” is a discipline form the variable part of the discipline block (modules) within the master training within 03.04.02 "Phisycs" (master program "Fumctional nanomaterials and advanced technologies"). The course is conducted during the second semester of the first term.

After attending the course, the students are expected to

Know underlying physics of the microscopes and sample requirements for the instruments, discussed within the course. 

Be able to determine the type of microscope most suited for solving the particular research problems and balance the possibilities and limitations of the instrument with the experimental requirements. 

Possess the necessary skills of wide-field transmitted light and fluorescent microscope manual operation and adjustment.