Excitation-scan mirror array system advancements to hyperspectral imaging applications

Parker, Marina; Browning, Craig M.; Mayes, Sam; Deal, Joshua; Gunn-Mayes, Samantha; Rich, Thomas C.; Leavesley, Silas J.

doi:10.1117/12.2546485

Cited by 2 publications

(1 citation statement)

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“…Compared with the previous TFTF-based approach, the LED-based HIFEX system provides greatly improved wavelength switching speeds. An optical geometry was developed 21 , 22 to allow for up to 16 predetermined excitation wavelengths while providing output through a standard liquid-light guide that was coupled to the microscope. To our knowledge, this is the first time that a multifaceted mirror geometry has been used to combine output from up to 16 single-band LEDs for spectral illumination.…”

Section: Introductionmentioning

confidence: 99%

Multifaceted mirror array illuminator for fluorescence excitation-scanning spectral imaging microscopy

et al. 2023

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. Significance Hyperspectral imaging (HSI) technologies offer great potential in fluorescence microscopy for multiplexed imaging, autofluorescence removal, and analysis of autofluorescent molecules. However, there are also associated trade-offs when implementing HSI in fluorescence microscopy systems, such as decreased acquisition speed, resolution, or field-of-view due to the need to acquire spectral information in addition to spatial information. The vast majority of HSI fluorescence microscopy systems provide spectral discrimination by filtering or dispersing the fluorescence emission, which may result in loss of emitted fluorescence signal due to optical filters, dispersive optics, or supporting optics, such as slits and collimators. Technologies that scan the fluorescence excitation spectrum may offer an approach to mitigate some of these trade-offs by decreasing the complexity of the emission light path. Aim We describe the development of an optical technique for hyperspectral imaging fluorescence excitation-scanning (HIFEX) on a microscope system. Approach The approach is based on the design of an array of wavelength-dependent light emitting diodes (LEDs) and a unique beam combining system that uses a multifurcated mirror. The system was modeled and optimized using optical ray trace simulations, and a prototype was built and coupled to an inverted microscope platform. The prototype system was calibrated, and initial feasibility testing was performed by imaging multilabel slide preparations. Results We present results from optical ray trace simulations, prototyping, calibration, and feasibility testing of the system. Results indicate that the system can discriminate between at least six fluorescent labels and autofluorescence and that the approach can provide decreased wavelength switching times, in comparison with mechanically tuned filters. Conclusions We anticipate that LED-based HIFEX microscopy may provide improved performance for time-dependent and photosensitive assays.

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Section: Introductionmentioning

confidence: 99%