FLIM

Abberior Instruments introduce FLIM upgrade for STED microscopes

 Headline FIG small 650

Abberior Instruments STED microscope now available with full Fluorescence Lifetime IMaging (FLIM) functionality!

The new Abberior Instruments comprehensive FLIM package upgrade offers the following key features:

  • STED FLIM imaging in combination with our pulsed STED lasers
  • Online display of STED FLIM images during the measurement
  • Online lifetime calculation and/or dye separation based on lifetime data
  • Simultaneous FLIM acquisition in up to 4 spectral channels (based on Becker & Hickl SPCM hardware)
  • Full software integration into Imspector including evaluation functionality (also live evaluation of data)

Continue below to see a selection of images generated using the Abberior STED FLIM technique.

Tubu VIM Confocal 600Tubu VIM STED FLIM 1 600

Confocal and STED FLIM image of mammalian cells labelled with tubulin/ Atto647N and vimentin/ Abberior STAR 635P.  

 DNA Lam Conf FLIM 600DNA Lam STED FLIM small 600

Confocal FLIM and STED FLIM image of mammalian cells labeled with antibodies against lamin/ Atto647N and DNA/ Abberior STAR635P.  

 TOM NUP STED 600

STED FLIM image of mammalian cells labeled with antibodies against Tom20/ Atto647N and NUP153/ Abberior STAR635P. Note, that nuclear pore (NUP) complex subunits appear to be localized in the cytoplasm, which is due to the NUP import pathway into the nuclear membrane.

 Tubu VIM STED FLIM 2 600Tubu VIM STED Separation small 600

STED FLIM image of tubulin labeled with Atto647N and vimentin labeled with Abberior STAR 635P. The two labels (Atto647N and Abberior STAR 635P) have the same excitation spectra, but different lifetimes. Tubulin (red, short lifetime) and vimentin (green, long lifetime) can successfully be separated in the STED FLIM recording via their lifetimes.  

 Fig0 Conf FLIM SMALL b

Confocal FLIM image of a Convallaria sp., rhizome cross section.  

Fig1 2cConf FLIM SMALL small

Two color confocal FLIM images of a Convallaria sp., rhizome cross section. Note that the
channels were recorded simultaneously.

Pollen1Pollen2 b

Pollen3 bPollen4 b

Confocal FLIM images of pollen grains.

Abberior specifications:

  • FLIM (Fluorescence lifetime imaging)
  • STED FLIM: FLIM imaging in combination with STED imaging
  • TCSPC photon streams can be auto-saved in OBF or B+H format. OBF streams can be loaded into MATLAB, Python (Numpy), C/C++, ImageJ including meta-data for off-line analysis
  • FCS (fluorescence correlation spectroscopy), scanning-FCS and other single-molecule spectroscopy applications through off-line analysis
  • Online lifetime calculation and/or dye separation based on lifetimes
  • Online display of lifetime images during the measurement
  • Software gating in combination with STED
  • Simultaneous FLIM/TCSPC acquisition in up to 4 spectral channels
  • 10 MHz combined saturated count rate (per module)
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