Fuorescence And Confocal Microscopy Flashcards Preview

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Flashcards in Fuorescence And Confocal Microscopy Deck (9):
0

Advantages of fluorescence microscopy

- very sensitive detection of labeling,
- high structural resolution
- multiple labeling is easy
- ideal for studying dynamic processes

1

Explain stokes shift

The energy difference between the peaks of the absorbance and the peak of the emission spectrum

2

Problems with fluorescence microscopy

- bleed through
- blur
- bleaching
- photo toxicity
- background fluorescence

3

How to remove blur

Optically
Hybrid
Computationally
Combination

4

Methods for countering photobleaching

Scan for shorter time
Higher magnification
Wide emission filters
Reduce excitation intensity
Use anti fade reagents

5

Describe 2nd harmonic imaging

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

Applications of fluorescent proteins

- organelle specific proteins
- sub cellular localization
- co localization analysis of signaling molecules
- whole tissue analysis
- time lapse studies of live specimens
- Physiological measurements

7

FRAP

Fluorescence recovery after photobleaching

8

FRET

Fluorescence resonance energy transfer