Introduction:To study the cell and its components, it is necessary to visualize and visualize them in detail. In this hands-on lesson we will examine several microscopic techniques that visualize cellular structures and identify their characteristics. Because most cells are very small, they cannot be seen with the naked eye and therefore must be magnified. Light microscopy was first used to magnify the image of cells using dyes. However, some tissues and subcellular structures are too small to be seen even under a light microscope. Therefore another technique was found to view the cell in more detail. To study the smallest features of the cell, electron microscopy is used. Electron microscopy uses a beam of electrons to view the sample. Electron microscopy can only magnify thin structures, so fluorescence microscopy is used to visualize thicker structures. Fluorescence microscopy visualizes light-emitting structures by allowing light to pass through the sample. Methods Question 1: Briefly describe how immune staining with an antibody is performed. Immune staining with an antibody is performed by binding the antibody to a specific antigen. This is done by adding a fluorescent dye to a thin section of a tissue or cell. A secondary antibody is coupled to a fluorescent label (1) that can attach to the primary antibody and enhance the light signal. Then, increase visibility.Question 2: Fluorescence/Immunofluorescence Complete Figure 1 below (i.e. draw the light paths).Figure 1. Diagram showing the light path for both A. Transmitted light illumination and B. Fluorescent illumination ( Epi-illumination) in an inverted microscope. The red lines indicate the path of the light that reaches the center of the sheet and its functions. References: (1) Dr. Potonik S, Lecture 2 Notes, BIOL2319 “Visualizing cells”, 4 March 2014(2) http://orthomolecular.org/nutrients/glicogen.html(3) http://www.plosone. org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0077166(4) http: //www.princeton.edu/~achaney/tmve/wiki100k/docs/Barr_body.html(5) http://cellmontage2 .cira.kyoto-u.ac.jp/cgi-bin/cellinf.cgi?cellname=basophil (6) http://www.google.com.au/url?sa=t&rct=j&q=&esrc=s&frm=1&source =web&cd=3&ved=0CD8QFjAC&url=http%3A%2F%2Fwww.ocr.org.uk%2FImages %2F140893-unit-r074-calculating-magnification-lesson-element-teacher-instructions.pdf&ei=wwUpU63QGMqQlQXn8oGgBg&usg=AFQjCNGRnI5IFykp3 Qso-eihUA0_x7gl_w(7 ) https://www.jic.ac.uk/microscopy/intro_LM.html(8) http ://www.ammrf.org.au/myscope/sem/ background/(9) http://en.wikipedia. org/wiki/High-length_transmission_electron_microscopy