The Colourful World of Fluorescent Proteins
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The document summarizes fluorescent proteins, including green fluorescent protein (GFP) and its variants. It describes how fluorescent proteins enable detection of proteins, organelles and cells. Their applications include studying cancer, cell migration and connections. It also discusses photoconvertible proteins like EosFP, which changes fluorescence from green to red upon exposure to light, enabling detection of movement. Fluorescent proteins assist in creating high-resolution images using techniques like laser scanning confocal microscopy and photoactivated localization microscopy.
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- 1. The Colourful World of Fluorescent Proteins Presented by: Caroline Sepiol Federici et al., 2012
- 2. The Green Fluorescent Protein and Variants life.bio.sunysb.edu; brainwindows.wordpress.com; Chalfie et al., 1994; bcgsc.ca; Zimmer, 2009
- 3. A “Small” List of Variants http://www.uniprot.org/uniprot/?query=gfP&sort=score
- 4. The Applications of Fluorescent Proteins Arun Sampathkumar and Elliot Meyerowitz (unpublished) ; Livet et al., 2007 Fluorescent proteins enable the detection of proteins, organelles and cells Their applications are nearly endless. For instance they can be used to study cancer, cell migration and connections. They also can be used in both plants and animal cells.
- 5. The “Brainbow” Experiment Litchman et al. 2008
- 6. The “Brainbow” Experiment Litchman et al. 2008
- 7. Why Fluorescent Proteins Glow? Heim et al., 1994
- 8. Why Fluorescent Proteins Glow? Webb and Miller, 2012
- 9. Photoconvertible Fluorescent Proteins Shaner et al., 2007; Wiedenmann et al., 2006; Chudakov et al., 2004
- 10. Photoactivable Fluorescent Proteins Piatkevich and Verkhusha, 2010; Shaner et al., 2007; Victoria et al., 2010
- 11. Photoswitchable Fluorescent Proteins Shaner et al., 2007; Fedor et al., 2010
- 12. *NEW* Fluorescent Timers • Limited group of proteins • Slowly changes emission from green to red or blue to red over a time period Subach et al., 2009
- 13. The Ideal Candidate Checklist Expressed efficiently Bright enough to be detected and imaged Stable fluorescence and integrity No interaction with the protein of interest Minimal interference Shaner et al., 2005
- 14. EosFP and it’s two colours This fluorescent proteins is photoconvertible and it is found in stony coral Lobophyllia hemprichii . The protein in its native state exhibits a green fluorescence with an emission maximum at 516 nm. Following violet- blue light exposure, the chromophore undergoes an irreversible photoconvertible reaction and emits a red fluorescence at 581 nm Wiedenmann et al., 2004; Nienhaus et al., 2005
- 15. The Ideal Candidate Checklist For EosFP: Expressed efficiently Bright enough to be detected and imaged Stable fluorescence and integrity No interaction with the protein of interest Minimal interference
- 16. How EosFP Detects Movement and Creates Images Laser Scanning Confocal Microscopy McKinney et al., 2009
- 17. How EosFP Enables the Detection of Movement and Assists in Creating Images Fluorescent Recovery after Photo-bleaching Mathur et al., 2010
- 18. How EosFP Enables the Detection of Movement and Assists in Creating Images Photoactivated Localization Microscopy How it works: http://zeiss-campus.magnet.fsu.edu/articles/superresolution/palm/introduction.html
- 19. How EosFP Enables the Detection of Movement and Assists in Creating Images How EosFP Detects Movement and Creates Images Betzig et al., 2006
- 20. Acknowledgements Dr. Kohalmi Travis Howes Andra Jejeran Dr. Menassa
Editor's Notes
- 3D projection of a Z-stack of confocal images of fluorescent Arabidopsis thaliana leaf cells. A green fluorescent marker is used to mark cells in blue and autofluorescence from chloroplasts is shown in red.
- A depth color-coded set of aligned confocal Z-sections of microtubule arrays (mCherry:TUA5) in Arabidopsis leaf epidermal pavement cells, which exhibit complex polarity.
- Rabbit Kidneys Cells; PS-CFP-hDAT tracking within filopodia of HEK293 cells
- opossum kidney epithelial cells.
- opossum kidney cells. Dronpa The movie is represented by the row of images from left to right: red fluorescence (in red pseudocolor) image; yellow fluorescence (in green pseudocolor) image; and the overlay between the two. The movie contains 6 frames per one cycle in the total of 11 cycles. The data refer to the bacterial colony expressing rsTagRFP-EYFP fusion. Note that the rsTagRFP acceptor red fluorescence kindling is accompanied by the EYFP donor yellow fluorescence quenching.
- created three fluorescent timers based on mCherry with slow, medium and fast maturation rates. In this image, the clock numbers are colonies of bacteria that have been expressing the fast fluorescent timer protein for the number of hours indicated by the clock numeral, yielding a real-time display of the conversion from blue to red fluorescence
- HeLa cells