Fluorescence Microscope Illumination Filter with Segmented Passbands
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Solution Overview
Problem
Current illumination systems for fluorescence microscopes, which use single bandpass filters, limit image quality by making non-fluorescent tissues appear black and restrict observation to a single fluorophore, requiring filter set switching for multiple fluorophores.
Innovation Solution
An illumination system with an emission spectrum including fluorescence excitation wavelengths and visible-light background illumination wavelengths, using an illumination filter with higher transmissivity for excitation wavelengths than background illumination wavelengths, allowing simultaneous observation of fluorophores and tissue with attenuated background illumination.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If single bandpass filters are used for illumination, then fluorescence excitation is achieved with high contrast, but non-fluorescent tissues appear black and only single fluorophore observation is possible
Solution Approach 1:
The illumination filter is segmented into multiple passbands: a first passband for fluorescence excitation wavelengths and a second passband for visible-light background illumination wavelengths. This segmentation allows simultaneous transmission of excitation light for fluorescence and visible light for tissue illumination, enabling both fluorophore observation and non-fluorescent tissue visualization without requiring filter switching.
Solution Approach 2:
The illumination filter performs multiple functions simultaneously: it transmits fluorescence excitation wavelengths to excite fluorophores, transmits visible-light background illumination wavelengths to illuminate non-fluorescent tissues, and blocks other wavelengths. This multi-functionality eliminates the need to switch between different filter sets for different observation modes.
2Reliability
If single bandpass filters are used for illumination, then fluorescence excitation is optimized, but the system requires filter set switching for multiple fluorophores
Solution Approach 1:
The illumination filter enables continuous observation without filter switching by providing uninterrupted transmission of both fluorescence excitation wavelengths and visible-light background illumination wavelengths. The system maintains continuous illumination and observation capability, eliminating the interruptions and time loss associated with switching between different filter sets for different fluorophores.
3Illumination intensity
If visible-light background illumination is added to fluorescence excitation, then tissue illumination is improved, but background light intensity may overwhelm fluorescence signal
Solution Approach 1:
The illumination filter applies different transmissivity characteristics to different wavelength regions: the first passband has high transmissivity for fluorescence excitation wavelengths to ensure strong fluorescence excitation, while the second passband has lower transmissivity for visible-light background illumination wavelengths to prevent overwhelming the fluorescence signal. This local quality differentiation optimizes both tissue illumination and fluorescence detection.
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
Enables the simultaneous observation of fluorescent and non-fluorescent tissues with comparable intensity, improving image quality and allowing observation of multiple fluorophores without filter switching.
Implementation Method 1
an illumination device (9), the illumination device having an emission spectrum which comprises fluorescence excitation wavelengths of the at least one fluorophore
Implementation Method 2
an illumination filter (41) for arrangement between the illumination device and the object, the illumination filter having at least one fluorescence excitation passband (93) which comprises at least the fluorescence excitation wavelengths and at least one background illumination passband (95) which comprises the visible-light background illumination wavelengths
Data Source
AI summary
The invention relates to an illumination system (10) for a fluorescence microscope (3) for observation of an object (17) containing at least one fluorophore (19), to a microscope (1) and to a microscope method for illumination of an object (17) comprising at least one fluorophore (19). Solutions of the art have the disadvantage that orientation within an object (17) is difficult and visibility of fluorescing regions of the object (17) is non satisfying. The inventive illumination system (10) improves the visibility of an object (17) under study by comprising an illumination device (9), with an emission spectrum (22) which includes fluorescent excitation wavelengths (23) of the at least one fluorophore (19) and visible-light background wavelengths (25), further comprising a illumination filter (41) having at least one fluorescence excitation passband (93) and at least one background illumination passband (95), wherein the transmissivity (88)/width (92) of the fluorescence excitation passband (93) is larger/smaller than the transmissivity (88)/width (92) of the background illumination passband (95). The inventive illumination system (10) is adapted to perform the inventive microscope method.