FLIM and Beyond: A Multimodal Approach to Confocal Spectroscopy

Fluorescence microscopy is a cornerstone of biophotonics, and fluorescence lifetime imaging microscopy (FLIM) offers powerful contrast by mapping variations in fluorescence lifetime across a sample. Because lifetime is independent of fluorophore concentration and highly sensitive to the local microenvironment, FLIM is valuable for detecting chemical and conformational changes, probing molecular interactions, and distinguishing species with overlapping emission spectra. As a result, FLIM has become an increasingly important tool in biological imaging.

Integrating FLIM with complementary spectroscopic techniques further enhances the analytical capability. Spectral photoluminescence (PL) imaging and Raman microscopy probe electronic and vibrational structure and are widely used on confocal Raman systems. Extending these systems include nonlinear and time resolved modalities, such as second harmonic generation (SHG), two photon excited fluorescence (2PEF), and phosphorescence lifetime imaging, enables multimodal workflows that capture structural, chemical, electronic, and dynamic information within one instrument.

This talk will demonstrate how FLIM can be seamlessly integrated into a confocal Raman microscope and how a suite of complementary spectroscopic techniques can be combined to support comprehensive multimodal imaging. The optical and detection design strategies enabling pulsed and continuous-wave excitation, time resolved photon counting, and spectrally resolved detection within a single instrument will be discussed. Using biological examples, we will highlight how multimodal spectroscopic imaging enhances contrast mechanisms, expands sample compatibility, and maximizes the information gained from every measurement.

About the presenter
Iona Hill, Ph.D., completed her doctorate at the University of Strathclyde, UK, where she specialized in optical medical imaging, using Raman spectroscopy to assess radiation therapy treatment response. She then continued her work in cancer treatment response during a postdoctoral position at UTHealth Houston, USA, investigating combination radiation therapy approaches. In 2024, Iona joined Edinburgh Instruments as an applications scientist, where she supports researchers using the company’s Raman and multimodal confocal microscopy platforms and applies her expertise in spectroscopic imaging across a wide range of biological and materials science applications.