Renishaw is pleased to introduce fluorescence lifetime imaging microscopy (FLIM) functionality to its inVia™ confocal Raman microscope. In partnership with Becker & Hickl GmbH, the pioneers in time-correlated single photon counting (TCSPC), the system combines each Raman and FLIM in a single instrument. This integration can overlay each FLIM and Raman images with pixel-to-pixel correlation, thus increasing understanding of samples across a spread of applications.

Image Credit: Renishaw

FLIM imaging

Within the system, when light from a pulsed laser interacts with a fluorophore, the fluorescence lifetime is measured using a complicated detector and high-speed electronics. This fluorescence lifetime is the characteristic time a fluorophore stays in an excited electronic state before returning to the bottom state by emitting a photon. The lifetime of fluorescence decay is sensitive to the molecular environment and the resulting changes in molecular conformation. The FLIM technique might be used to review a spread of properties from pH/ion/oxygen concentration to molecular binding.

Combined FLIM with Raman microscopy

FLIM and Raman microscopy are complementary methods for label-free imaging of biological cells and tissue sections. FLIM provides insight into molecular interactions and might be performed not less than 10× faster than Raman imaging. Raman spectroscopy boasts high chemical specificity and high-resolution chemical images. This mix of techniques is good for rapid identification of regions of interest by FLIM, followed by chemical and structural characterisation by Raman imaging.

For FLIM data evaluation, comprehensive software from Becker & Hickl is included with multi-exponential, incomplete and shifted-component evaluation models. The software also includes proprietary phasor evaluation to tell apart lifetime populations. Fluorescence lifetime and Raman images might be directly overlaid because of using the highly accurate Renishaw MS30 high speed encoded stage for each imaging modalities.

“We’re pleased to announce an extra enhancement to our award-winning inVia Raman microscope. Our collaboration with the leaders in FLIM technology, Becker and Hickl, has produced a combined FLIM Raman microscope resulting in exciting latest application areas.” 

David Reece, Renishaw Spectroscopy Business Development Manager

For further information on fluorescence lifetime imaging microscopy (FLIM): Fluorescence lifetime imaging microscopy (FLIM) (renishaw.com)

For further information on Renishaw Raman Spectroscopy: www.renishaw.com/raman