The course will focus on three common methods used in biology: tissue culture, protein analysis by Western blot and basic light microscopy. The principles of each method will be discussed as well as the applications and important parameters to consider. Students will have a hands on practice on each method, and will get useful links and details on available facilities for advanced applications of these methods.
-Background on tissue culture: What is tissue culture and what type of questions can be addressed by cultured cells, advantages and limitations. Different types of cultures will be discussed, and the main characteristics of each will be explained (cell lines, primary cells, adherent and non-adherent cells, mammalian vs. insect and bacteria)
-General handling of cultured cells: How to grow, maintain, subculture, count, freeze and thaw. Principles and important practicalities of working with cultured cells.
-Manipulation to culture cell: Expression and deletion of specific genes, tagging proteins, and protein production by cultured cells.
-Hands-on: Work in a sterile environment, subculture cells, live cell labeling, and counting cells.
-Background: Principles of protein separation on gels, different types of gels, staining/labeling that can be done directly on a gel (TCE, coomassie blue, fluorescent labeling)
-Western blot: Uses, benefits and limitations. Different types of transfer and membranes, how to choose which one to use. How to visualize target protein, and ways of imaging. Links to available scanners in Weizmann.
-Hands-on: Run cell lysate on SDS-PAGE gel, practice different gel staining methods (TCE vs. coomassie blue), compare wet and semi dry transfer, and western blot.
Light microscopy: (I will not cover topics that are covered in the advanced course)
-Background: Basic principles of bright field microscopy and current uses. Principles of fluorescent microscopy, microscope components and optics.
-Cell labeling: Live vs. fixed cell labeling. Tagged proteins, fluorescently labeled antibodies and cellular markers. Limitations of light microscopy and common uses.
-Review what types of microscopes are available in Weizmann, specificities of each and links to each.
-Hands-on: Operating basic bright field/fluorescent microscope. Imaging of unlabeled live cells, work with different lenses (long vs. short distance), imaging fluorescently labelled cells.