Learning Outcomes

Core Module

The core module focuses on fluorescence microscopy because it is the central tool of our lab.

At the end of this training, a student should be able to:

Identify the components of an epifluorescence microscope

• The students should know the names of the components when pointed to on the training microscope
• The students should be able to identify (approximately for hidden components) where on a real microscope the same components are located
• The students should be able to state the purpose of each component
• The students should be able to sketch a basic epifluorescence microscope from memory

Assess the relative brightnesses of fluorescence signals

• The students should be able to acquire flourescence images at high SNR without saturation by tuning the microscope parameters while at the scope
• The students should know how to remove the effects of camera offset
• The students should know how to perform background subtraction and identify when it is necessary to do so
• The students should be able to estimate how much brighter the signal is in one fluorescence channel vs. another
• The students should be able to estimate relative amounts of proteins, etc. by correcting fluorescence brightness for quantities such as quantum yield and absorption cross sections

Measure the degree of fluorescence cross talk between channels

• The students should be able to define what fluorescence cross talk is and explain why it is problematic
• The students should be able to perform the necessary measurements to assess cross talk
• The students should be able to explain what steps one might take to alleviate the effects of cross talk