BMS 534

Introduction to Confocal Microscopy & Image Analysis

2 credit hours 
1 credit offered in Spring (theory)
1 credit offered Maymester (tutorial/practical)

instructor
	J. P. Robinson

prerequisites
	None

description
	Confocal Microscopy provides a powerful tool for evaluation of the
 3D structure of a variety of tissues and biological samples, as well as some
 non-biological samples.  In addition, using the multiple excitation capacity
 of these instruments it is possible to identify multiple sites of interest
 within a single sample and to reproduce theses data using image reconstruction
 techniques.  Live cells can also be evaluated using these technologies to
 monitor calcium fluxes, pH changes or production of oxygen and nitrogen 
radicals using fluorescent probes.
	Participants will become familiar with the rudiments of confocal 
and fluorescent microscopes, and associated technologies.  They will learn
 the fundamentals of fluorescence detection and develop a keen understanding
 of the use of and availability of a variety of fluorescent probes across 
the UV and visible spectrum.  They will also be introduced to basic image
 analysis techniques.  Participants will learn to use both the UV/visible 
confocal scope and the associated operating software.  They will have the 
opportunity to run the instrument in small groups and will be required to 
produce images demonstrating their capability.  They will also learn the 
use of the real-time linescan confocal scope using video acquisition software
 including Universal Imaging Metamorph software.  At the end of the course,
 all participants should be capable of understanding the theory and should
 be capable of essentially operating instrumentation currently housed in 
the Cytometry Laboratories at Purdue University.

text

outline
	    Lecture Topic (spring 1 lecture per week)
1. The Principles of Fluorescence Microscopy
2. The Principles of Confocal Microscopy
3. Intro to the Bio-Rad MRC 1024 Confocal System
4. Fluorescent Probes
5. Sample Preparation for Confocal Microscopy
6. Images Formats and Image Manipulations
7. Principles of 2D image analysis - Morphometry (I)
8. Applications of Morphometry (II)
9. Intro to the Bio-Rad DVC Viewscan System
10. 3D reconstruction of biological events
11. Edge Detection & Object Classification
12. Calcium Imaging Applications
13. Visualization techniques
14. confocal Microscopy of Neuroscience
15. Advanced Applications of Imaging
                   -------------------------------------------



Practical Course: Maymester
8 hours per week for 4 weeks
-------------------------
1. Introduction to the Confocal System
2. Imaging using transmitted light
3. Imaging using fluorescence
4. Time course measurements
5. 3D imaging and reconstruction
---------------------------------------

For further information regarding the Biomedical Engineering Program at Purdue University contact the Biomedical Engineering Graduate Office at (317) 494-5730 bmeprogram@ecn.purdue.edu