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Introduction to Confocal Microscopy
& Image Analysis
BMS 524, 2 Credit Course Lectures: Spring 1997, Practicals Maymester 1997
Professor: J.Paul Robinson
Ideal for Graduate Students, Seniors - Faculty & Staff invited to monitor
Course 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 these 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 microcsopes, 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.
The course is STRICTLY LIMITED because of the need for practical participation.
This is the first such course offered
at Purdue University. Acceptance into the course will be at the instructor’s discretion; however, current graduate students
will be given priority. There are NO PREREQUISITES.
Course Structure:
Instructor: J. Paul Robinson
Lectures: 16 hours (Spring semester)
Practical Demonstrations: 10 hours (Maymester)
Practical Participation: 20 hours (Maymester)
Duration: January 13-June, 1996
Tentative Schedule (may change): Lectures will be Wednesdays 11:30 - 12:20 during Spring semester. Practicals will be given during Maymester: practical demonstrations will be Tuesday 1-3 pm; Thursday 1-3pm; practical (hands-on) sessions will be Tuesday and Thursday 3-5 pm.
This course is offered through the Purdue University Cytometry Laboratories which is
under the Department of Basic Medical
Sciences in the School of Veterinary Medicine. Funding for the Course is provided by
the Department, the Vet School, and via the
Vice President's Reinvestment Fund.
Lectures:
(Given by Dr. J. Paul Robinson unless otherwise indicated)
1. The Principles of Fluorescence Microscopy
2. The Principles of Confocal Microscopy
3. Fluorescent Probes
4. Sample Preparation for Confocal Microscopy
5. Image Formats and Image Manipulations
6. Introduction to the Bio-Rad MRC 1024 Confocal System
7. Introduction to the Bio-Rad DVC Viewscan System
8. Principles of 2D image analysis - Morphometry(I) (Dr. John Turek)
9. Applications of Morphometry (II) (Dr. John Turek)
10. 3D Reconstruction
11. Confocal Microscopy for Neuroscience (Dr. Terry Powley)
12. Live Cell Applications
13. Calcium Imaging Applications (Dr. Ken Robinson)
14. Applications in Immunology (TBA)
15. Advanced Applications of Imaging: 2 Photon Imaging (TBA)
16. Course Overview
Grading: Final Examination: 65% Practicals - preparation & analysis of appropriate images: 35%
Back to Purdue University Cytometry Labs Homepage
For information contact J.Paul Robinson, Ph.D.,
robinson@flowcyt.cyto.purdue.edu
Director PUCL, 1515 Hansen B050, Purdue University, West Lafayette, IN
47907-1515, U.S.A. Phone: (317) 494-0757; Fax: (317) 494-0517; robinson@flowcyt.cyto.purdue.edu
CD ROM Vol 2 was produced by staff at the Purdue University Cytometry Laboratories
and distributed free of charge as an educational service to the cytometry community.
If you have any comments please direct them to Dr. J. Paul Robinson, Professor & Director, PUCL,
Purdue University, West Lafayette, IN 47907. Phone:(317) 494-0757; FAX (317) 494-0517; Web http://www.cyto.purdue.edu
EMAIL robinson@flowcyt.cyto.purdue.edu
