Principles and Applications of Fluorescence Microscopy
Instructor: Marin Manuel, PhD, University of Rhode Island
Location: URI
Session 1: June 5 – June 7 ( Lab: Room 405 / Lecture Room: Avedisian Rm 244L)
Session 2: June 17 – June 19 ( Lab: Room 405 / Lecture Room: Avedisian Rm 244L)
Course Overview
This WDT Module aims to train students in the use of fluorescence microscopy, confocal microscopy and image analysis. The module includes lectures on the theory behind each of these
techniques, as well as hands-on observation of biological samples on a state-of-the-art microscope. The module is intended for students who are new to microscopy and will provide foundational knowledge that can be directly applied to many fields of research where fluorescence microscopes are in use. The module will be completed over 2.5 days and participants who complete the module will receive an RI-INBRE Certificate of Completion.
Learning Outcomes
- Learn the principles of fluorescence microscopy
- Understand the difference between excitation and emission filters
- Choose the appropriate fluorochromes for each application
- Learn the theory behind confocal microscopy
- Learn operation of the Nikon Eclipse Ti2 inverted confocal microscope
- Analyze imaging data using the commonly used image processing software, ImageJ
Lab Report
Students will be expected to maintain detailed laboratory notes to include:
- Material and fluorophores used
- Imaging parameters
- Data analysis protocols (e.g., workflows, software used, etc.)
- Additional Information
Resources Used in this Module:
- Slide Decks for Lectures
- A computer running Windows/macOS/Linux for data processing
- Image processing program ImageJ
Timeline
| Day 1 | Day 2 | Day 3 | |
|---|---|---|---|
| 9:00 AM-10:00 AM | Introduction to Fluorescence and fluorophores (lecture) | Introduction to confocal microscopy (lecture) | Data Analysis - General Principles (Lecture/Hands-On) |
| 10:00 AM-11:00 AM | Visit of the INBRE microscopy facility | Introduction to confocal microscopy (lecture) | Data Analysis - use of ImageJ to analyze images acquired during the first two days (Hands-on) |
| 11:00 AM-12:00 PM | Introduction to widefield microscopy (lecture) | Hands-on manipulation on the Nikon Eclipse Ti2 microscope | Data Analysis - use of ImageJ to analyze images acquired during the first two days (Hands-on) |
| 12:00 - 1:00 PM | Break | Break | WDT Survey and Certificate Distribution |
| 1:00 PM-2:00 PM | Hands-on manipulation on the Nikon Eclipse Ti2 microscope | Hands-on manipulation on the Nikon Eclipse Ti2 microscope | |
| 2:00 PM-3:00 PM | Hands-on manipulation on the Nikon Eclipse Ti2 microscope | Hands-on manipulation on the Nikon Eclipse Ti2 microscope | |
| 3:00 PM-4:00 PM | Hands-on manipulation on the Nikon Eclipse Ti2 microscope | Hands-on manipulation on the Nikon Eclipse Ti2 microscope) |