BIOLOGY 557: ULTRASTRUCTURAL TECHNIQUES Fall 2010 Use of the Facility is predicated on successful completion of a portfolio of images utilizing these techniques and instruments. All images generated as part of this class are the property of the EM Facility. This laboratory is intended to train students in electron and light microscopic techniques and will take a minimum average of 8 hours per week in the laboratory. Students are not permitted to work in the lab outside of 8:30 am-5 p.m. M-F without express permission of Dr. Barlow. Students will learn how to prepare grids to support samples in the TEM negatively stain small samples preserve and embed samples for slicing prepare knives used in slicing create surface replicas of samples record and process images align and use the light and transmission electron microscope slice both thick and thin slices for light and electron microscopic examination Text: ELECTRON MICROSCOPY, J. Bozzola and L.Russell 2nd edition, Jones & Bartlett (1999) EM Facility instrument tutorials GRADING: Late assignments incur a grading penalty: Time Sheets All work hours in lab 5% Every 2nd and 4th Friday, 3 pm (Includes weekly meetings) Lab report I Negative staining 15% Friday noon, October 15 Lab report II Thick sections 15% Friday noon, October 29 Lab report III Thin slicing 20% Tuesday 1pm, November 30 Lab report IV Shadowing 15% Friday noon, December 3 Lab report V Class presentation 10% Tuesday class, December 7 Lab report VI Tomography 20% Friday noon, December 10 Lab Practicals TEM solo exam Trimmed Blocks Thick sections Light scopes & cameras Credit Credit Credit Credit No later than October 1 No later than October 8 No later than October 15 No later than October 22 Lab activities will consist primarily of labeled micrographs highlighting each of the techniques discussed. Additional guidelines will be given out in class. Lab work will be graded on attendance, quality, timeliness, and adherence to guidelines All techniques have sections in the Bozollo and Russel Book. Read the pertinent sections in the book BEFORE you come to the lab.Tentative schedule: Week Date Exercise To be performed BEFORE lab Week 1 8/31 Introduction to laboratory Week 2 9/7 Grids, negative stain, TEM operation Watch grid loading insertion, neg. stain, grids videos Read TEM operation handout Week 3 9/14 TEM operation, camera Read TEM/camera operation handout Week 4 9/21 Knives, trimming blocks, thick sections Watch Sectioning videos Week 5 9/28 Thick sections Prepare 2 trimmed blocks Week 6 10/5 Light microscopes, digital cameras Week 7 10/12 Fixation Read Fixation handout Week 8 10/19 Thin sectioning Week 9 10/26 Tomography Read Tomo handouts Week 10 11/2 Tomography Read IMOD handouts Week 11 11/9 Shadowing Read Shadowing handout Week 12 11/16 SEM/STEM demonstration Week 13 11/23 Open Lab Week 14 11/30 Open Lab Week 15 12/7 Class