Instructor: Dr. Bob Youker (Western Carolina University)

In this lecture/lab course, students will learn how to prepare their field samples for brightfield and fluorescence microscopic examination. Students will also learn how microscopes generate images, the experimental conditions needed for optimal imaging of their specimens, and learn basic image processing techniques for analysis. The course will be a combination of lecture and hands on specimen preparation and imaging (lab). Students can either provide specimens from their field research, or work with the instructor to collect rotifers and other aquatic micro-invertebrates from the field to use in the lab portion.   

Lab time will be centered on learning the parts and functions of modern brightfield and fluorescence (widefield & confocal) microscopes, preparing specimens (e.g., staining with fluorescent dyes), and acquiring images of their specimens for analysis. Students will learn to acquire images using a widefield fluorescence microscope and take a “field trip” to WCU to use the state-of-the-art Leica Stellaris 5 confocal microscope. We will also discuss some other modern approaches to fluorescence microscopy, such as electron and super-resolution microscopies. 

Each student will prepare a final report that compares brightfield to fluorescence images taken of their samples. The student will explain in detail the features observed using both methodologies and the advantages and disadvantages of each method. The student will also give a short oral presentation on one microscopic technique not covered in the course and this topic will be selected from a list provided by the instructor.

Pre-requisites: Introductory biology sequence