2 Photon Basics
What are the limitations to confocal imaging?
Light scatter, photobleaching and photodamage
What animal was GFP isolated in?
Jellyfish
Name one of the motivations listed for the paper?
Relationship between firing and detection is unclear
Undetected APs are common
Requires large neuron population
Calcium dependent fluorescence transfer can vary neuron to neuron
What is 2 photon Fluorescence?
Imaging technique that uses two photons of longer wavelength (lower light energy) to generate higher energy fluorescence
What year was GFP discovered? What year did scientists started using it in labs commercially?
1971. 1990s.
Name one type of neurons were used in this paper?
Emx1-s
tetO-s
Cux2-f
Emx1-f
What is the unit of time used to trap photons?
A femto second
These scientists were awarded the Nobel price in 2008 for the discovery of GFP
Roger Y. Tsien, Osamu Shimomura, and Martin Chalfie
How many mouse lines were used in this paper?
6
What type of diagram shows energy transitions between excitation and emission?
Jablonski Diagram
What is the structure of GCaMP?
GFP- Calmodulin (CaM)-M13 (peptide from myosin light chain Kinase)
What was one of the algorithms used for AP detection in this paper?
NND, L0, MLspike
Name 2 of the 4 benefits to 2 photon imaging?
Is GCamp intensiometric? Or Ratiometric? Why?
Intensiometric –
Measures one event during any given time
Which algorithm performed slightly better?
MLspike