Development of a compressed sensing platform for High-speed Hyperspectral FLIM

Session

Poster Session Two

Authors

Mr Harry Bakhshi (1), Dr. Anneliese Jarman (1), Dr. Simon Poland (1)

Affiliations

1. King’s College London

Keywords

FLIM, SPAD, Hyperspectral, Compressed Sensing, 3D imaging, Cancer imaging

Abstract text

We propose a hyperspectral FLIM platform utilising a SPAD CMOS line sensor detection architecture, in parallel with compressed sensing, for increased image acquisition speed over a broad wavelength range. This platform is well-suited to multi-label samples prepared to investigate cancer cell biology, in both 2D and 3D, with complex fluorescence spectral signatures. 

Fluorescence lifetime imaging (FLIM) is a key microscopy technique used to measure local variations in biological cells and tissues and can report on many characteristics within the cellular environment. When used in conjunction with multispectral acquisition, it enables precise characterisation and quantification of both extrinsic fluorescent probes and intrinsic fluorescent metabolic markers from complex overlapping fluorescence emission spectra (Datta, R. et al., 2020). We report on the development of a hyperspectral FLIM microscopy system which may acquire multiple frames per second, with lower saturation effects and higher lifetimes detected, for deeper, faster and more selective 3D cancer model interrogation. This is achievable due to advances in histogramming on-chip (Erdogan, A. T. et al., 2019) and via superposition of binary holograms using DMD (Digital Micromirror Device) projection. Projected DMD patterns such as a [Walsh-] Hadamard may then be exploited by a compressed sensing algorithm, to increase resolution for the generated frame (Ochoa, M. et al., 2018), and speed of acquisition. 

Hyperspectral FLIM with compressed sensing meets the need to monitor dynamic biological events in 3D cancer models, in real time, over physiologically-relevant short timescales.

References

Datta, Rupsa, Tiffany M. Heaster, Joe T. Sharick, Amani A. Gillette, and Melissa C. Skala. “Fluorescence Lifetime Imaging Microscopy: Fundamentals and Advances in Instrumentation, Analysis, and Applications.” Journal of Biomedical Optics 25, no. 7 (July 2020): 071203. https://doi.org/10.1117/1.JBO.25.7.071203.

Erdogan, Ahmet T., Richard Walker, Neil Finlayson, Nikola Krstajić, Gareth Williams, John Girkin, and Robert Henderson. “A CMOS SPAD Line Sensor With Per-Pixel Histogramming TDC for Time-Resolved Multispectral Imaging.” IEEE Journal of Solid-State Circuits 54, no. 6 (June 2019): 1705–19. https://doi.org/10.1109/JSSC.2019.2894355.

Ochoa, M., Q. Pian, R. Yao, N. Ducros, and X. Intes. “Assessing Patterns for Compressive Fluorescence Lifetime Imaging.” Optics Letters 43, no. 18 (September 15, 2018): 4370–73. https://doi.org/10.1364/OL.43.004370.