High-speed compressed-sensing fluorescence lifetime imaging microscopy of live cells

Ma, Yayao; Lee, Young Jae; Best-Popescu, Catherine; Gao, Liang

doi:10.1101/2020.07.16.205161

Cited by 6 publications

(8 citation statements)

References 60 publications

(39 reference statements)

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“…Such an end-to-end training and datadriven approach prevents any empirical bias that may negatively impact the design. We envision that such an approach will enable many challenging applications such as superresolution imaging [23,[42][43][44][45][46], 3D imaging [41,[47][48][49], and high-speed computational camera [50][51][52].…”

Section: F Advantage Of End-to-end Optimized Auto-encoder and Its Application In Future Imaging Systemsmentioning

confidence: 99%

Deep compressed imaging via optimized pattern scanning

Zhang

Yang

2021

Photon. Res.

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The need for high-speed imaging in applications such as biomedicine, surveillance and consumer electronics has called for new developments of imaging systems. While the industrial effort continuously pushes the advance of silicon focal plane array image sensors, imaging through a single-pixel detector has gained significant interests thanks to the development of computational algorithms. Here, we present a new imaging modality, Deep Compressed Imaging via Optimized-Pattern Scanning (DeCIOPS), which can significantly increase the acquisition speed for a singledetector-based imaging system. We project and scan an illumination pattern across the object and collect the sampling signal with a single-pixel detector. We develop an innovative end-to-end optimized auto-encoder, using a deep neural network and compressed sensing algorithm, to optimize the illumination pattern, which allows us to reconstruct faithfully the image from a small number of samples, and with a high frame rate. Compared with the conventional switching-mask based single-pixel camera and point scanning imaging systems, our method achieves a much higher imaging speed, while retaining a similar imaging quality. We experimentally validated this imaging modality in the settings of both continuous-wave (CW) illumination and pulsed light illumination and showed high-quality image reconstructions with a high compressed sampling rate. This new compressed sensing modality could be widely applied in different imaging systems, enabling new applications which require high imaging speed.

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Section: F Advantage Of End-to-end Optimized Auto-encoder and Its Application In Future Imaging Systemsmentioning

confidence: 99%

Deep compressed imaging via optimized pattern scanning

Zhang

Yang

2021

Photon. Res.

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Section: Introductionmentioning

confidence: 99%

“…Studies to date involving smFRET and FLIM either exploited the measurements of FLT in multiple color channels in order to enhance the robustness or accuracy of the measurement or used this information to gain knowledge on the photophysical behavior of the dyes (Kaye et al, 2017) (Ho ¨fig et al, 2014 (Sheng et al, 2008). Other studies used laser scanning-based FLT measurements or large-area FLIM for live cell FLIM (Oleksiievets et al, 2020;Raspe et al, 2016) (Blacker et al, 2014;Grecco et al, 2010;Lagarto et al, 2020;Ma et al, 2021;Scipioni et al, 2021). FLIM, optical nanopore sensing, as well as other applications of FLT may nevertheless benefit from the ability to resolve multiple fluorescence species in each of the excitation/emission channels, as a way to further increase the methods' multiplexibility, while preserving single-molecule resolution.…”

Section: Introductionmentioning

confidence: 99%

Lifetime-based analysis of binary fluorophores mixtures in the low photon count limit

Nasser

Meller²

2022

iScience

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Summary Single biomolecule sensing often requires the quantification of multiple fluorescent species. Here, we theoretically and experimentally use time-resolved fluorescence via Time Correlated Single Photon Counting (TCSPC) to accurately quantify fluorescent species with similar chromatic signatures. A modified maximum likelihood estimator is introduced to include two fluorophore species, with compensation of the instrument response function. We apply this algorithm to simulated data of a simplified two-fluorescent species model, as well as to experimental data of fluorophores' mixtures and to a model protein, doubly labeled with different fluorophores' ratio. We show that 100 to 200 photons per fluorophore, in a 10-ms timescale, are sufficient to provide an accurate estimation of the dyes' ratio on the model protein. Our results provide estimation for the desired photon integration time toward implementation of TCSPC in systems with fast occurring events, such as translocation of biomolecules through nanopores or single-molecule burst analyses experiments.

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“…An improved CUP variant was proposed, called the trillion-frame-per-second CUP (TCUP) [28], working at a time resolution of 100 fs by including an unsheared view recorded by an additional camera. Recently, CUP has been further improved for various applications, including high-dimensional optical imaging [29], ultraviolet imaging [30], ultrafast electron diffraction imaging [31], phase-sensitive imaging [32] and fluorescence lifetime imaging [33][34][35]. For more information about CUP, please refer to the review paper [36].…”

Section: Introductionmentioning

confidence: 99%

Theoretical investigations of a modified compressed ultrafast photography method suitable for single-shot fluorescence lifetime imaging

Tian

2021

Appl. Opt.

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A single-shot fluorescence lifetime imaging (FLIM) method based on the compressed ultrafast photography (CUP) is proposed, named space-restricted CUP (srCUP). srCUP is suitable for imaging objects moving slowly (< ∼ 150/M mm/s, M is the magnification of the objective lens) in the field of view with the intensity changing within nanoseconds in a measurement window around 10 ns. We used synthetic datasets to explore the performances of srCUP compared with CUP and TCUP (a variant of CUP). srCUP not only provides superior reconstruction performances, but its reconstruction time is also two-and threefold faster than CUP and TCUP, respectively. The lifetime determination performances were assessed by estimating lifetime components, amplitude-and intensity-weighted average lifetimes (τ A and τ I ) with the reconstructed scenes using the least square method based on a bi-exponential model. srCUP has the best accuracy and precision for lifetime determinations with a relative bias less than 7% and a coefficient of variation less than 7% for τ A and a relative bias less than 10% and a coefficient of variation less than 11% for τ I .

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High-speed compressed-sensing fluorescence lifetime imaging microscopy of live cells

Cited by 6 publications

References 60 publications

Deep compressed imaging via optimized pattern scanning

Deep compressed imaging via optimized pattern scanning

Lifetime-based analysis of binary fluorophores mixtures in the low photon count limit

Theoretical investigations of a modified compressed ultrafast photography method suitable for single-shot fluorescence lifetime imaging

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