Novel Multifocal fluorescence lifetime imaging platform


Super-resolution and localization fluorescence microscopy techniques have attracted considerable attention in the past decade in particular (including the Nobel Prize in Chemistry, 2014), as they allow for localization of fluorophores on length scales below the optical diffraction limit, and elucidation of nanometer scale structural features in biological samples.


Academics at King’s College London have developed a novel imaging prototype with multi-beam excitation and detection capabilities for use in confocal  and multiphoton fluorescence imaging which can be attached to any existing microscope.

This development we believe is a paradigm shift over existing fluorescence lifetime imaging tools and provides the user with the ability to monitor protein-protein interactions in a high content screening environment.

Fluorescent microscopy has proved to be an invaluable tool for live scientists in the understanding of biological interactions. It is currently a multi-billion pound industry which continues to expand and evolve rapidly as new requirements and imaging technologies are being developed. Compared the number of life scientists performing intensity based fluorescent imaging, the Fluorescent lifetime imaging community is composed of a relatively small number of dedicated groups. This is due in part to the intricacy involved in biological sample preparation as well as an understanding in the complex analysis required to analyse acquired datasets.



Technical Status


Proof of principle: The technology has been demonstrated experimentally. 



IP Status


UK priority application



Commercial Status


Industry partners are being sought for commercial development of the technology with a view to licensing this technology.



Key features

  • Retro fit  - It can be attached onto any existing microscope system.
  • Speed – The acquisition rates faster than any other FLIM based system.
  • Accuracy -  This provides unparalleled temporal measurement accuracy provided by TCSPC to measure lifetime.
  • Cost Effective – This technology has the potential to be much less expensive than existing FLIM based systems.



Market Opportunity


This system is primarily being developed for high content screening of pathological tissue samples to monitor protein-protein interactions but could be used in general microscopy applications  and in flow cytometry. 

The system would be primarily used to image FRET interactions for high speed pathological screening but can be used either in in-vivo, in-vitro and ex-vivo imaging situations. This includes protein-protein homo- or hetero-dimer interactions and with FRET biosensors. The system can also be used with fluorescence lifetime probes to monitor localised environment variations. We envisage the device as a self contained module could be attached onto a number of existing microscopy systems such as confocal, multiphoton, endoscopic, High content screening and flow imaging.

The set-up provides a platform for future improvements in speed and signal-to-noise by increasing the number of beams or using smaller area SPADs. Such advances have the potential to transform time-resolved multiphoton imaging applications in a range of biological systems. Now we can utilise unparalleled temporal measurement accuracy provided by TCSPC to measure lifetime with high frame rates to image complex live cell interactions dynamically.


There are a number of commercially developed systems currently available which operate in either the time-domain (using TCSPC or time gating) or the frequency domain (phase modulation). Becker & Hickl and Picoquant offer complete TCSPC based laser scanning confocal FLIM systems which can be attached onto any existing microscope. Several companies (including Princeton instruments, Stanford computer optics)  market time-gated image intensified Charge Coupled Devices (ICCD) for both widefield fluorescence and spinning disk confocal set-ups. To date all of these techniques are expensive (>£100k) and as such are cost prohibitive for many life scientists. Typical laser scanning TCSPC, whilst unrivalled in temporal and spatial accuracy, are simply too slow to monitor dynamic biological events. Time-gating or phase modulation techniques offer much greater speeds but suffer from systematic error due to measurement methodologies incorporated.


With our multifocal TCSPC based FLIM technique there is no longer a need to compromise between temporal accuracy, speed and spatial resolution. Translating this technology into a self-contained cost-effective imaging platform for use in high content screening is an attractive proposition and has the potential to expand the existing market due to its ease of use and practicality.


Patent Information:
Physical Sciences
For Information, Contact:
Mugdha Joshi
IP & Licensing Manager
King's College London
Simon Ameer-Beg
James Levitt