Fluorescent principle. Fluorescent microscope DAPI.

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Presentation transcript:

Fluorescent principle

Fluorescent microscope

DAPI

FITC

Alexa 488

Tetramethylrhodamine/TRITC

Rhodamin

Rhodamin Red-X

Alexa 568

Alexa 647

Texas-Red and Texas-Red-X

Alexa 594

Cy-Series

GFP and GFPuv

E-GFP's and DsRed

Table of Fluorescent Proteins

What is green fluorescent protein (GFP) ? GFP is a 238-amino acid protein originally isolated from the bioluminescent jellyfish Aequoria victoria Can be expressed in any organism without additional cofactors

Structurally, GFP represents a new motif which has been named the “β-can” The “can” consists of 11 β-strands and a central α-helix Structure of GFP

GFP-Fluorophore Fluorescence is mediated by a fluorophore located at the center of the protein

The fluorophore is derived from the tripeptide -Ser 65 -Tyr 66 - Gly 67 - Formation of the fluorophore occurs in an autocatalytic manner Structure of the GFP-fluorophore

Light sources for Fluorescence

Filtersets and GFP's

Quantum efficiency of Chip

New Filterset

New Filterset-other flourochromes

Old Filtersets

Old filtersets- other flurochromes

Antibodies used for fluorescence

Laser and Fluorochromes