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Quantitative Fluorescence Microscopy

A national techniques course in fluorescence microscopy and imaging

Date:  May 16–22, 2014
Location:   MDI Biological Laboratory

Overview

fluorescence

This one week, intensive microscopy course will cover all aspects of the technology from the principals of fluorescence imaging to multidimensional imaging in living cells. Lectures dealing with the theory, mechanics, and application of fluorescent imaging methods will be intermingled with extended laboratory practicals in which students will be encouraged to use their own specimens, to optimize the utility of the course. Our goal will be to provide students with the knowledge and expertise to implement cutting edge microscopic methods within their own laboratories.

Registration

Apply Now

Contact

Simon C. Watkins, Ph.D., Course Director
Professor, Cell Biology & Physiology
University of Pittsburgh School of Medicine
412-648-3051
swatkins@pitt.edu

 

Judi Medlin
Education Coordinator
207-288-9880 x102

Tuition

$3,100 (room and board included)

Rationale

The power and capability of the light microscope has increased dramatically over the last few years. In fact light microscopy is now becoming of central utility in all fields of biological research including cell biology, physiology and molecular biology. This change has been driven by the need to study increasingly sophisticated problems at high spatial and temporal resolution. The advances, principally in the field of fluorescence microscopy have been dependant on the development of entirely new microscopic methodologies coupled to the use of fluorescent proteins, new fluorescent dye technologies, highly sensitive detectors and inexpensive powerful computers.

Our goal will be to provide students with the knowledge and expertise to implement cutting edge microscopic methods within their own laboratories. In contrast with other currently available courses it will focus exclusively on the theory, potential application and implementation of fluorescent imaging techniques, and perhaps more importantly this will be couched in the real world application of biological experimentation.

This one week, intensive course will cover all aspects of the technology from the principals of fluorescence imaging to multidimensional imaging in living cells. Lectures dealing with the theory, mechanics, and application of fluorescent imaging methods will be intermingled with extended laboratory practicals in which students will be encouraged to use their own specimens, to optimize the utility of the course.

Teaching Faculty

Simon C. Watkins, Ph.D.
Professor, Cell Biology & Physiology
University of Pittsburgh School of Medicine
Pittsburgh, PA 15261 USA
Phone:(412)648-3051
Fax: 648-8330
swatkins@pitt.edu

David W. Piston, Ph.D.
Professor of Molecular Physiology and Biophysics
Associate Professor of Physics
Vanderbilt Medical Center
1211 22nd Ave S
Nashville, TN 37232

Claudette M. St. Croix, Ph.D.
Assistant Professor
Department of Environmental and Occupational Health University of Pittsburgh Graduate School of Public Health
s220 BST 3500 Terrace Street
Pittsburgh, PA 15219

Anne Kenworthy, Ph.D.
Assistant Professor
Vanderbilt University
Nashville, TN

Gert-Jan Kremers, Ph.D.
Imaging Centre Erasmus MC
Rotterdam, The Netherlands

Industrial Faculty

 
lecture/lab topics and times subject to change
Friday, May 16, 2014
4:00 pm Arrival
Check in to on or off-campus housing, keys at MDIBL Dining Hall

6:00-7:30pm

 

Welcome dinner and social
MDIBL DIning Hall

7:30pm Course introduction
MDIBL Conference Center
Saturday, May 17, 2014
(self serve breakfast every day in the Dining Hall)
9:00-9:30 am Introduction
Welcome, Introductions, Define Course Goals
Watkins/Piston
9:30-10:30 am The Microscope 1
From Van Leeuwenhoek to Digital Imaging
Watkins
10:30-11:00 am Break
11:00-12:00 pm The Microscope 2
Transmitted Light Contrast in Microscopy
Piston
12:00-1:00 pm Lunch
1:00-3:00 pm LAB 1
TRANSMITTED LIGHT MICROSCOPY
3:00-4:00 pm The Microscope 3
Fluorescence Microscopy and Quantitation
Piston
4:00-4:30 pm
The Microscope 4
Optical Filters for Flourescemce Microscopy
Stanley
4:30-5:00 pm Break
5:00-6:00 pm Digital Imaging 1
Digital Imaging, Resolution and Detectors
Watkins
6:00-7:00 pm Dinner
7:00-8:00 pm Digital Imaging 2
Noise and Brightness Demonstration
Piston/Watkins
8:00-11:00 pm LAB 2
STANDARD FLUORESCENCE MICROSCOPY
Sunday, May 18, 2014
9:00-9:45 am Optical Sectioning 1
Introduction to Confocal Microscopes
St. Croix
9:45-10:30 am

Optical Sectioning 2
Confocal Detectors and "Pseudo-confocals"
Piston

10:30-11:00 pm Break
11:00-12:00 pm Optical Sectioning 3
FRAP, Photoactivation, TIRF
Kenworthy
12:00-1:00 pm Lunch
1:00-4:30 pm
LAB 3
CONFOCAL MICROSCOPY
4:30-5:30 pm Tips and Tricks
Digital Filters
Watkins
6:00-7:00 pm Dinner
7:00-11:00 pm LAB 4
IMAGE PROCESSING & FEATURE EXTRACTION
Monday, May 19, 2014
9:00-9:30 am Image Processing 1
Practical Digital Imaging
Watkins
9:30-10:30 am Image Processing 2
Deconvolution
Piston
10:30-11:00 am Break
11:00-12:00 pm Optical Sectioning 4
3D Methods, Microfluorimetry Spectral Imaging
St. Croix
12:00 noon Course Photo
12:00-1:00 pm Lunch
1:00pm-4:00 pm LAB 5
MULTICOLOR 3-D IMAGING
4:00-5:30 pm Tips and Tricks
Cell Labeling, Antibodies, Practical Aspects
Watkins
6:00 pm
Dinner, evening off
Tuesday, May 20, 2014
9:00-10:00 am Live Cell Imaging 1
Structured Illumination, Super-Resolution
Kenworthy
10:00 am - 110:30 am
Image Processing 3
TConsiderations for Image Quantitation
Piston
10:30-11:00am Break
11:00-12:00 Live cell Imaging 2
GFP
Kremers
12:00-1:00 pm Lunch
1:00-5:00 pm LAB 6
DECONVOLUTION, APOTOME, FRAP
5:00-6:00 pm Tips and Tricks
Live Cell Imaging Design
Watkins
6:00-7:00 pm Dinner
7:00-8:00 pm LAB 7
TIRF, FRAP, PHOTOACTIVATION
Wednesday, May 21, 2014
9:00-9:45 am Live Cell Imaging 3
Multicolor and Ratio Imaging
St. Croix
9:45-10:30 AM

Optical Sectioning 5
Multi-Photon Microscopy
Piston

10:30-11:00 pm Break
11:00-12:00 pm Live Cell Imaging 4
FRET and Live Cell Imaging
Kenworthy
12:00-1:00 pm Lunch
1:00-5:00 pm LAB 8
FRET IMAGING: FIXED AND LIVE CELLS
5:00-5:30 pm LAB 9
LIVE CELL CHAMBERS, SETUP COLLABORATIVE GROUPS BASED ON STUDENT INTERESTS
6:00-7:00 pm Dinner
7:00-11:00 pm LAB 10
TIME-LAPSE LIVE CELL IMAGING, QUANTITATION, RATIO IMAGING METHODS TRACKING, STUDENT EXPERIMENTS
Thursday, May 22, 2014
9:00-9:30 am Live Cell Imaging 5
Comparison of Optical Sectioning Approaches
Piston
9:30-10:30 am Image Processing 4
Advanced Image Processing
Watkins
10:30-11:00 am Break
11:00-2:00 pm LAB 11
Advanced Image Analysis, COLLECT DATA, PREPARE PRESENTATIONS
12:00-1:00 pm Lunch
2:00-4:00 pm Results!
Student Presentations
4:00-4:30 pm Mechanics
How Do I Get Money to Pay for All This Stuff?
Watkins
4:30-5:00 pm Summing up
Q&A, Student Feedback
Faculty
5:30 pm Lobster Bake
Lab Beach, weather permitting
Friday, May 23, 2014
9:00 am Departure
Check out of on-campus housing by 9 am

Housing: On-campus housing is available in single and double-accommodation dormitory rooms and cottages. The course fee includes the cost of on-campus housing.

Travel, Shuttles, and Public Transportation: Detailed travel information, including airport and ground transportation suggestions, is available on the MDIBL Travel page.

Most course participants will be housed on-campus and daily transportation will not be necessary.  Directions to MDIBL are also available.

The Bar Harbor Chamber of Commerce has information regarding local attractions, including Acadia National Park.

Full Tuition fellowships have been generously provided by Chroma Technologies, Nikon, Carl Zeiss Microimaging, and the Maine INBRE.

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