Multi-Beam FLIM Illumination for Deep Tissue Penetration
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Solution Overview
Problem
Fluorescence Lifetime Imaging Microscopy (FLIM) technologies face challenges with low fluorescence intensity and poor tissue penetration, particularly in deeper tissue layers, leading to false results and misdiagnosis in cancer diagnosis.
Innovation Solution
A method of evenly illuminating tissue samples in wide field FLIM using multiple light beams or a square wave excitation, which distributes excitation photons more evenly across the sample, increasing the probability of deeper tissue penetration and improving diagnostic accuracy.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If traditional short pulse excitation techniques are used in wide field FLIM, then the equipment complexity is reduced and operation is simplified, but the fluorescence intensity is low and tissue penetration is poor, leading to false results and misdiagnosis
Solution Approach 1:
The illumination system is segmented into multiple independent laser beams (at least two beams) that can be directed at different angles to illuminate the tissue sample from multiple directions. This segmentation allows each beam to contribute to illuminating different depths of the tissue, improving overall penetration and diagnostic accuracy without requiring a single complex high-power source
Solution Approach 2:
The patent introduces angular diversity by illuminating the tissue sample from multiple angles rather than from a single direction. This dimensional change in illumination geometry enables deeper tissue penetration and improves fluorescence signal collection from various tissue depths, resolving the contradiction between diagnostic accuracy and system complexity
2Illumination intensity
If single beam illumination is used, then the system is simpler, but deeper tissue layers are not visible, causing false results and misdiagnosis
Solution Approach 1:
Multiple laser beams illuminating from different angles are merged in their effect on the tissue sample, with each beam contributing to the overall fluorescence excitation. The combination of beams from multiple angles produces enhanced fluorescence intensity and improved tissue penetration compared to a single beam, while maintaining manageable system complexity through the use of standard optical components
Solution Approach 2:
The patent employs periodic modulation of the laser beams at different frequencies, allowing for frequency-domain FLIM measurements. This periodic action enables the extraction of fluorescence lifetime information while maintaining adequate signal intensity from multiple illumination paths, resolving the contradiction between signal strength and system complexity
3Length of stationary object
If excitation light is directed at one side only, then the illumination setup is simpler, but penetration into deeper tissue layers is insufficient
Solution Approach 1:
The patent illuminates the tissue sample from opposite sides (first side and second side) simultaneously or sequentially. This inverted approach of bidirectional illumination allows excitation light to penetrate from both directions, effectively doubling the penetration depth capability and enabling visualization of deeper tissue layers that would be inaccessible with unidirectional illumination
Solution Approach 2:
Different regions of the tissue sample receive optimized illumination from specific directions. By directing beams at different angles, the system provides locally optimized excitation for different tissue depths and regions, improving overall penetration depth while maintaining a relatively simple illumination configuration using standard optical elements
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
This approach enhances the visibility of deeper tissue layers, increases fluorescence photon count, and provides more accurate and reliable medical diagnoses by capturing images from multiple surfaces or using extended excitation waves, overcoming limitations of traditional short pulse techniques.
Implementation Method 1
FLIM captures a fluorescence lifetime which reflects how long the dye stays in the excited state before returning to the ground state by emitting a fluorescence photon
Implementation Method 2
The emission of a fluorescence photon from a fluorophore does not, however, always occurs at an exact time after excitation and a distribution of time is observed, showing an exponential decay function
Data Source
AI summary
A method of evenly illuminating a tissue sample in a wide field Fluorescence Lifetime Imaging Microscopy (FLIM) and a system for practice thereof. Particularly provided here is a method of evenly illuminating a slide containing a tissue sample in a wide field light source by (i) illuminating the slide containing the tissue sample from more than one side, or (ii) illuminating the slide containing the tissue sample using a square wave excitation, or (iii) illuminating the slide containing the tissue sample using a combination of (i) and (ii) and capturing an individual wide field image or a plurality of wide field images of the tissue sample for accurate medical diagnosis of a medical condition.