Presentation is loading. Please wait.

Presentation is loading. Please wait.

Powerpoint Templates Page 1 Powerpoint Templates Spectroscopic Microscopy.

Published byMorgan Carson Modified over 8 years ago

Similar presentations

Presentation on theme: "Powerpoint Templates Page 1 Powerpoint Templates Spectroscopic Microscopy."— Presentation transcript:

1 Powerpoint Templates Page 1 Powerpoint Templates Spectroscopic Microscopy

2 Powerpoint Templates Page 2

3 Powerpoint Templates Page 3 Limitation of light microscopy 5  m limit of resolution blue

4 Powerpoint Templates Page 4 Confocal Microscopy

5 Powerpoint Templates Page 5

6 Powerpoint Templates Page 6 Absorption Spectra of Fluors Commonly Conjugated to Secondary Antibodies Fluorochrome AbsorptionEmission Cascade Blue400420 Fluorescein494518 Rhodamine570590 Texas Red595615 Cy5650670

7 Powerpoint Templates Page 7 Colocalization of insulin and calcitonin receptor-like receptor Insulin-Cy5 CRLR-FITC Glucagon-Rhodamine

8 Powerpoint Templates Page 8

9 Powerpoint Templates Page 9

10 Powerpoint Templates Page 10 Fluorescence Resonance Energy Transfer FRET is the non-radioactive transfer of photon energy from an excited fluorophore (the donor) to another fluorophore (the acceptor) when both are located within close proximity (1-10nm). Using FRET one can resolve the realtive proximity of molecules beyond the optical limit of a light microscope to reveal (1) molecular interactions between two protein partners, (2) structural changes within one molecule (eg. enzymatic activity or DNA/RNA conformation), (3) ion concentrations using special FRET-tools like the CFP-YFP cameleon

11 Powerpoint Templates Page 11 No FRET Signal FRET Signal CFP is excited by light and emits light CFP is more than 10nm from YFP YFP is not excited and does not emit light CFP is excited by light and emits light CFP is in close proximity to YFP YFP emits light

12 Powerpoint Templates Page 12

13 Powerpoint Templates Page 13 FLIM (Fluorescence Lifetime Imaging Microscopy) measures the lifetime of the excited state (delay between excitation and emission) every fluorophore has a unique natural lifetime lifetime can be changed by the environment, such as: Ion concentration Oxygen concentration pH Protein-protein interactions ∆t=lifetime

14 Powerpoint Templates Page 14 FRAP (Fluorescence Recovery After Photobleaching)

15 Powerpoint Templates Page 15

Download ppt "Powerpoint Templates Page 1 Powerpoint Templates Spectroscopic Microscopy."

Similar presentations

Part 1: The root of all evil Part 2: Fluorescence microscopy

Part 1: The root of all evil Part 2: Fluorescence microscopy
Fluorescence and Confocal Microscopy

Fluorescence and Confocal Microscopy
Page 1 Klaus Suhling Department of Physics Kings College London Strand London WC2R 2LS UK Fluorescence Lifetime Imaging (FLIM) of molecular rotors maps.

Page 1 Klaus Suhling Department of Physics Kings College London Strand London WC2R 2LS UK Fluorescence Lifetime Imaging (FLIM) of molecular rotors maps.
Victor Sourjik ZMBH, University of Heidelberg

Victor Sourjik ZMBH, University of Heidelberg
Flow Cytometric Analysis of FRET to Study the Interaction Between CFP- and YFP-Tagged Proteins David Stepensky.

Flow Cytometric Analysis of FRET to Study the Interaction Between CFP- and YFP-Tagged Proteins David Stepensky.
Fluorescence Microscopy Wolfgang Graier F-actin NFkB (activation by H 2 O 2 ) Pictures: W.F Graier, MBC & MB, Graz, Austria.

Fluorescence Microscopy Wolfgang Graier F-actin NFkB (activation by H 2 O 2 ) Pictures: W.F Graier, MBC & MB, Graz, Austria.
Fluorescence and Confocal Microscopy Yvona Ward Cell and Cancer Biology Branch.

Fluorescence and Confocal Microscopy Yvona Ward Cell and Cancer Biology Branch.
Live cell imaging. Why live cell imaging? Live cell analysis provides direct spatial and temporal information Planning your experiment – The markers/fluorophores.

Live cell imaging. Why live cell imaging? Live cell analysis provides direct spatial and temporal information Planning your experiment – The markers/fluorophores.
Fluorophores bound to the specimen surface and those in the surrounding medium exist in an equilibrium state. When these molecules are excited and detected.

Fluorophores bound to the specimen surface and those in the surrounding medium exist in an equilibrium state. When these molecules are excited and detected.
Gert-Jan Kremers FRET and Live Cell Imaging Wednesday, May 21, QFM 2014.

Gert-Jan Kremers FRET and Live Cell Imaging Wednesday, May 21, QFM 2014.
Special Applications in Fluorescence Spectroscopy Miklós Nyitrai; 2007 March 14.

Special Applications in Fluorescence Spectroscopy Miklós Nyitrai; 2007 March 14.
Methods: Fluorescence Biochemistry 4000 Dr. Ute Kothe.

Methods: Fluorescence Biochemistry 4000 Dr. Ute Kothe.
Chemical Structure of the Chromophore Biosynthesis of the Chromophore Critcial dehyrogenation reaction to juxtapose aromatic group with imidazlinone.

Chemical Structure of the Chromophore Biosynthesis of the Chromophore Critcial dehyrogenation reaction to juxtapose aromatic group with imidazlinone.
Oligonucleotides – Primers and Probes by … as quality counts! Competence and Service in Molecular Biology metabion´s history.

Oligonucleotides – Primers and Probes by … as quality counts! Competence and Service in Molecular Biology metabion´s history.
The use of fluorescence resonance energy transfer (FRET) to study the interaction between CD147 and MCT1 The principle of FRET MCT1 CD147 s s s s N- terminus.

The use of fluorescence resonance energy transfer (FRET) to study the interaction between CD147 and MCT1 The principle of FRET MCT1 CD147 s s s s N- terminus.
Immunofluorescence Microscopy. Immunofluorescence Microscopy: When an antibody, or the antiimmunoglobulin antibody used to detect the antibody is labeled.

Immunofluorescence Microscopy. Immunofluorescence Microscopy: When an antibody, or the antiimmunoglobulin antibody used to detect the antibody is labeled.
Microscopy is about a combination of resolution (seeing smaller and smaller things), and contrast (seeing what you want to see). Both aspects have recently.

Microscopy is about a combination of resolution (seeing smaller and smaller things), and contrast (seeing what you want to see). Both aspects have recently.
Microscopy is about a combination of resolution (seeing smaller and smaller things), and contrast (seeing what you want to see). Both aspects have recently.

Microscopy is about a combination of resolution (seeing smaller and smaller things), and contrast (seeing what you want to see). Both aspects have recently.
Spectroscopy of Proteins. Proteins The final product of the genes, translated form genes (mutation in gene leads to a mutated protein) Made of a verity.

Spectroscopy of Proteins. Proteins The final product of the genes, translated form genes (mutation in gene leads to a mutated protein) Made of a verity.
Study of Protein Association by Fluorescence-based Methods Kristin Michalski UWM RET Intern In association with Professor Vali Raicu.

Study of Protein Association by Fluorescence-based Methods Kristin Michalski UWM RET Intern In association with Professor Vali Raicu.

Similar presentations

Ads by Google