Fuorescence And Confocal Microscopy Flashcards
0
Q
Explain stokes shift
A
The energy difference between the peaks of the absorbance and the peak of the emission spectrum
1
Q
Advantages of fluorescence microscopy
A
- very sensitive detection of labeling,
- high structural resolution
- multiple labeling is easy
- ideal for studying dynamic processes
2
Q
Problems with fluorescence microscopy
A
- bleed through
- blur
- bleaching
- photo toxicity
- background fluorescence
3
Q
How to remove blur
A
Optically
Hybrid
Computationally
Combination
4
Q
Methods for countering photobleaching
A
Scan for shorter time Higher magnification Wide emission filters Reduce excitation intensity Use anti fade reagents
5
Q
Describe 2nd harmonic imaging
A
Irradiation with femtosecond laser results in forward scatter of light of double the frequency of the excitation light
Able to image structures closely index matched to surrounding media invisible to phase-contrast
6
Q
Applications of fluorescent proteins
A
- organelle specific proteins
- sub cellular localization
- co localization analysis of signaling molecules
- whole tissue analysis
- time lapse studies of live specimens
- Physiological measurements
7
Q
FRAP
A
Fluorescence recovery after photobleaching
8
Q
FRET
A
Fluorescence resonance energy transfer
