Course Syllabus Course Name: Cell Biology Laboratory First Year Available: Year 3 (Junior) Course Code: SCNS480 Subject Area: Natural Science Class Style: Laboratory Number of Credits: 1 Term(s) Offered: Spring Semesters Prerequisites: SCNS210 or SCNS310 Instructor(s): Michael Wolfgang Lassalle Course Description In this course the function and structure of cells will be employed. We will use the Light Microscope to quantify the differences between prokaryotic and eukaryotic cells, measure an Enzyme for the Krebs cycle, isolate Chloroplast and measure the Hill Reaction for Photosynthesis. Furthermore, we will study the membrane and Chromatin structure. The most typical Cell Biology technologies will be explained by hands‐on experience. Course Objectives The students will be able to distinguish prokaryotic and eukaryotic cells, understand the basic components, know how the cell produces energy and are able to connect changes of cell structure and function to disease. Grading Criteria Preparation for the experiment (25%); laboratory report and experiment (35%); laboratory notes (10%); activity in the laboratory (20%); final Group Poster Presentation of gained skills and knowledge (10%) Lab Groups will consist of three students. Each group has to submit the preparation notes (experimental design and procedure, theory for the experiment) a day before the laboratory class. Organization Classes will start with a 10‐minute discussion followed by the experiment time. Required Materials and Textbooks Cell Biology: Edition 2 Thomas D. Pollard William C. Earnshaw Jennifer Lippincott‐Schwartz April 26, 2007 Elsevier Health Sciences Medical Cell Biology: Edition 3 Steven R Goodman November 26, 2007 Academic Press Other texts will be posted online or will be available as copy. Rules Safety is mandatory and disregarding safety rules not an option. Therefore, safety rules have to be strictly obeyed. If a student uses spectroscopic equipment, or chemicals without receiving prior guidelines from the Instructor, or violates any other safety rules, he/she has will be dismissed from the laboratory and will obtain no credit for this course. Approximate teaching material costs incurred to each student Lab coat and goggles: 3000‐5000 yen. Miscellaneous materials used for experiments (cauliflower, spinach leaves): 200‐400 yen. Experiments and Readings WEEK 1 Introduction to the Light Microscope Quantify differences between prokaryotic and eukaryotic cells. Cell counting by the Hemocytometer • Reading: Cell Biology: Edition 2 Section I Chapter 1 Introduction to Cells Medical Cell Biology: Chapter 1 1‐23 Tools of the Cell Biologist WEEK 2 Energy producing Organelles (Mitochondria) Part 1 Isolation of Mitochondria • Reading: Cell Biology: Edition 2 Section VI Chapter 19 Mitochondria, Chloroplasts, Peroxisomes Medical Cell Biology: Chapter 4 Organelles Structure and Function http://www.columbia.edu/~crg2133/Files/CambridgeIA/Biology/CitricAcidCycle.pdf Rockstroh M., Mueller ST.A., Jende C., Kerzhner A., von Bergen M., Tomm J.M. (2011). Cell fractionation ‐ an important tool for compartment proteomics. Journal of Integrated Omics 1, 135‐
143. WEEK 3 Energy producing Organelles (Mitochondria) Part 2 Measurement of Succinate Dehydrogenase Activity from Isolated Mitochondria • Reading: Cell Biology: Edition 2 Section VI Chapter 19 Mitochondria, Chloroplasts, Peroxisomes Medical Cell Biology: Chapter 4 Organelles Structure and Function http://www.nobelprize.org/nobel_prizes/medicine/laureates/1953/krebs‐lecture.pdf WEEK 4 Energy producing Organelles (Chloroplast) Part 1 Chloroplast Isolation • Reading: Cell Biology: Edition 2 Section VI Chapter 19 Mitochondria, Chloroplasts, Peroxisomes Medical Cell Biology: Chapter 4 Organelles Structure and Function http://cbm.msoe.edu/markMyweb/SUN‐chlorophyllEbookWorking/ WEEK 5 Energy producing Organelles (Chloroplast) Part 2 Separation of Chlorophyll‐Protein Complexes by Electrophoresis •Reading: Cell Biology: Edition 2 Section VI Chapter 19 Mitochondria, Chloroplasts, Peroxisomes Medical Cell Biology: Chapter 4 Organelles Structure and Function Sraehlin L.A. (2003). Chloroplast structure: from chlorophyll granulates to supra‐architeture of thylakoid membranes. Photosynthesis Research. 76, 185‐196. Markwell J.P., Thornber J.P., and Boggs R.T. (1979). Higher plant chloroplasts: Evidence that all chlorophyll exists as chlorophyll‐protein complexes. Proc. Natl. Acad. Sci. USA 76, 1233‐1235. WEEK 6 Energy producing Organelles (Chloroplast) Part 3 The Hill Reaction in isolated Chloroplasts •Reading: Cell Biology: Edition 2 Section VI Chapter 19 Mitochondria, Chloroplasts, Peroxisomes Medical Cell Biology: Chapter 4 Organelles Structure and Function http://soilove.com/db/12/Hill%20reaction.pdf WEEK 7 Cell Biology and Enzymes Michaelis‐Menten Kinetics •Reading: Goldbeter A. (2013). Oscillatory enzyme reactions and Michaelis‐Menten kinetics. FEBS Letters 587, 2778‐2784. WEEK 8 The Cell Membrane Experiments related to Diffusion and Osmosis •Reading: Cell Biology: Edition 2 Section III Membrane Structure and Function Medical Cell Biology: Chapter 2 Cell Membranes 27‐56 http://www.biologycorner.com/bio1/notes_diffusion.html WEEK 9 Cell‐Biology and Disease Part 1 Enzyme‐linked immunosorbent assay (ELISA) •Reading: Medical Cell Biology: Chapter 9 Cell Cycle and Cancer 273‐276 Lequin R.M. (2005). Enzyme Immunoassay (EIA)/Enzyme‐Linked Immunosorbent Assay (ELISA). Clinical Chemistry 51, 2415‐2418. http://www.idexx.com/pubwebresources/pdf/en_us/livestock‐poultry/elisa‐technical‐guide.pdf WEEK 10 Protein fingerprinting and Isozymes (Cell‐Biology and Disease Part 2) Isozymes will be detected by fingerprinting •Reading: Spyrakis F., Singh R., Cozzini P., Campanini B., Salsi E., Felici P., Raboni S., Benedetti P., Crudani G., Kellogg G.E., Cook, P.F., and Mozzarelli A. (2013). Isozyme‐Specific Ligands for O‐acetylserine sulfhydrylase, a Novel Antibiotic Target. PLOS One 8, 1‐13 WEEK 11 Final test: Poster Presentation of gained skills