Spectral lifetime imaging microscopy (SLIM) simultaneously measures fluorescence lifetime and spectra for intrinsic and extrinsic fluorophores. This technique discriminates between multiple fluorophores and provides information about the microenvironment – ionic strength, bound state, oxygen concentration, etc. SLIM provides a “fingerprint” of a fluorophore allowing more accurate tracking by researchers, and fingerprints of intrinsic fluorophores such as NADH can provide metabolic maps of cells. The SLIM system at LOCI is fitted on a Nikon Eclipse TE2000U base. An excitation laser is tunable from approximately 700-980 nm. Three detectors allow for recording of emitted fluorescence. The Becker-Hickl PML-16 is a 16-channel spectral lifetime detector. Each channel acquires roughly 10 nm bins of the spectrum. The Hamamatsu H7422 gather traditional multiphoton intensity images and fluorescent lifetime data. The Hamamatsu MCP is an extremely sensitive detector used for fluorescent lifetime imaging. The system also contains a galvanometer scanning system from Cambridge Technologies, and all scans follow a raster pattern.
- Becker-Hickl PML-16
- Hamamatsu H7422
- Hamamatsu MCP
Semrock internal filters:
Maximum field of view: 10x objective, zoom 1.304 mm