NEWSLETTER 05-10-2006: whiteTIRF -Total Internal Reflection Fluorescence and FLIM: the new combination!
The Netherlands, 05-10-2006
TIRF -Total Internal Reflection Fluorescence and FLIM -Fluorescence Lifetime Imaging Microscopy can be combined,
even on the widefield fluorescence microscope!
TIRF -Total Internal Reflection Fluorescence microscopy facilitates extremely high-contrast visualisation and thereby high sensitivity of fluorescence near the cover glass (up to 100nm). The combination of TIRF -Total Internal Reflection Fluorescence with frequency domain FLIM -Fluorescence Lifetime Imaging Microscopy enables the study of the dynamic activity and interactions between different proteins in e.g. membranes of cells.
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| Lifetime image with TIRF: the details of proteins that are close to the coverslip are better visible than in the image without TIRF. | Lifetime image without TIRF (just epifluorescence): the image is more fuzzy than by TIRF, because the out-of-focus fluorescence is also visible. |
The images shown here are taken with the Nikon TE2000-U widefield microscope with white-TIRF illuminator, combined with the Lambert Instruments LIFA -Fluorescence lifetime imaging Attachment. The samples were kindly provided by Ms. S.E. Ledevedec, Leiden University, The Netherlands. As light source the modulated LED was used and as demonstrated here, enough intensity was generated to obtain fluorescence lifetime images with TIRF -Total Internal Reflection Fluorescence.
More information on TIRF -Total Internal Reflection Fluorescence and FLIM -Fluorescence Lifetime Imaging Microscopy can be found at our website: APPLICATIONS > CELL BIOLOGY > TIRF (Total Internal Reflection Fluorescence) -FLIM
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