Laboratory Techniques for Light Microscopy

At the end of the course, students should be able to: 1. discuss the basic principles and practical aspects of light microscopy; 2. demonstrate the use of the microscope with artificial or natural light; 3. explain the need for sample preparation and be able to mount and stain a specimen on a slide and prepare it for microscope viewing; 4. identify parts of a compound microscope and operate it effectively; 5. compare the terms magnification, contrast, and resolution; 6. make proper biological drawings of both and an animal and plant cell; 7. outline the processes involved in the preparation of tissue sections and explain the purpose of each of these processes; 8. review the principles of light microscopy and identify the major parts of the microscope; and 9. use the microscope to view slides of several different cell types, including the use of the oil immersion objective lens.

History of microscopy, composition of the microscope, contrast and resolution concepts, the practical step by step method of tissue processing for light microscope study shall be taught and demonstrated, transmitted light microscopy: brightfield, phase contrast, dark field, DIC and polarization microscopy, koller illumination, image acquisition, digital image acquisition, cameras, binning gain, principles of fluorescence microscopy, emission and excitation spectrum, stokes shift, basic techniques of optics, light & fluoresence microscopy - fluorescence filter cubes acquisition of simple and complex images, principles of confocal microscopy, the principles and techniques for the use of advance light microscopes will be taught and where possible demonstrated i.e. polarizing microscope, phase contrast microscope, interference microscope, dark-field Microscope, and Ultraviolet Microscope. Spectral microscope/spectral unmixing Sample preparation for microscopy: fixation, permeabilization, reducing auto fluorescence, making samples transparent, image processing.