Large field-of-view phase and fluorescence mesoscope with microscopic resolution

Kernier, Isaure De; Ali-Cherif, Anaïs; Rongeat, Nelly; Cioni, Olivier; Morales, Sophie; Savatier, Julien; Monneret, Serge; Blandin, Pierre

doi:10.1117/1.jbo.24.3.036501

Cited by 13 publications

(11 citation statements)

References 44 publications

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“…Surprisingly, this extremely simple way to produce QPI in microscopy is relatively new [28][29][30][31] . Mandula et al derived phase from defocus 28 as a simple and compact phase imaging microscope for long-term observation of non-absorbing biological samples as well as its combination with fluorescence microscopy in a single dual-mode imaging platform using a standard bright-field objective 29 .…”

Section: Abbreviationsmentioning

confidence: 99%

Single-shot wavelength-multiplexed phase microscopy under Gabor regime in a regular microscope embodiment

Micó

Rogalski

Picazo-Bueno

et al. 2023

Sci Rep

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Phase imaging microscopy under Gabor regime has been recently reported as an extremely simple, low cost and compact way to update a standard bright-field microscope with coherent sensing capabilities. By inserting coherent illumination in the microscope embodiment and producing a small defocus distance of the sample at the input plane, the digital sensor records an in-line Gabor hologram of the target sample, which is then numerically post-processed to finally achieve the sample’s quantitative phase information. However, the retrieved phase distribution is affected by the two well-known drawbacks when dealing with Gabor’s regime, that is, coherent noise and twin image disturbances. Here, we present a single-shot technique based on wavelength multiplexing for mitigating these two effects. A multi-illumination laser source (including 3 diode lasers) illuminates the sample and a color digital sensor (conventional RGB color camera) is used to record the wavelength-multiplexed Gabor hologram in a single exposure. The technique is completed by presenting a novel algorithm based on a modified Gerchberg–Saxton kernel to finally retrieve an enhanced quantitative phase image of the sample, enhanced in terms of coherent noise removal and twin image minimization. Experimental validations are performed in a regular Olympus BX-60 upright microscope using a 20X 0.46NA objective lens and considering static (resolution test targets) and dynamic (living spermatozoa) phase samples.

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Section: Abbreviationsmentioning

confidence: 99%

Single-shot wavelength-multiplexed phase microscopy under Gabor regime in a regular microscope embodiment

Micó

Rogalski

Picazo-Bueno

et al. 2023

Sci Rep

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show abstract

“…The issue gets even worse owing to the non-ideal optical performance of a scanning system. Particularly for a large imaging area in a mesoscopic imaging system (Sofroniew et al, 2016;Bumstead et al, 2018;Pacheco et al, 2017;Kernier et al, 2019), the optical aberrations (Egner and Hell, 2006) become prominent toward the edges and corners, unavoidably leading to non-uniform excitation and detection efficiencies across the FOV. This essentially further reduces the signal strengths of the weaker structures residing at the off-axis locations.…”

Section: Introductionmentioning

confidence: 99%

A rapid denoised contrast enhancement method digitally mimicking an adaptive illumination in submicron-resolution neuronal imaging

Borah

Sun

2022

iScience

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Optical neuronal imaging often shows ultrafine structures, such as a nerve fiber, coexisting with ultrabright structures, such as a soma with a substantially higher fluorescence-protein concentration. Owing to experimental and environmental factors, a laser-scanning multiphoton optical microscope (MPM) often encounters a high-frequency background noise that might contaminate such weak-intensity ultrafine neuronal structures. A straightforward contrast enhancement often leads to the saturation of the brighter ones, and might further amplify the high-frequency background noise. We report a digital approach called rapid denoised contrast enhancement (DCE), which digitally mimics a hardware-based adaptive/controlled illumination technique by means of digitally optimizing the signal strengths and hence the visibility of such weak-intensity structures while mostly preventing the saturation of the brightest ones. With large field-of-view (FOV) two-photon excitation fluorescence (TPEF) neuronal imaging, we validate the effectiveness of DCE over state-ofthe-art digital image processing algorithms. With compute-unified-device-architecture (CUDA)-acceleration, a real-time DCE is further enabled with a reduced time complexity.

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“…The issue gets even worse due to non-ideal optical performance of a scanning system. Particularly for a large imaging area in a mesoscopic imaging system [12][13][14][15], the optical aberrations [16] become prominent towards the edges and corners, unavoidably leading to non-uniform excitation and detection efficiencies across the FOV. This essentially further reduces the signal strengths of the weaker structures residing at the off-axis locations.…”

Section: Introductionmentioning

confidence: 99%

Real-time Noise-suppressed Wide-Dynamic-Range Compression in Ultrahigh-Resolution Neuronal Imaging

Sun

2021

Preprint

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With a limited dynamic range of an imaging system, there are always regions with signal intensities comparable to the noise level, if the signal intensity distribution is close to or even wider than the available dynamic range. Optical brain/neuronal imaging is such a case where weak-intensity ultrafine structures, such as, nerve fibers, dendrites and dendritic spines, often coexist with ultrabright structures, such as, somas. A high fluorescence-protein concentration makes the soma order-of-magnitude brighter than the adjacent ultrafine structures resulting in an ultra-wide dynamic range. A straightforward enhancement of the weak-intensity structures often leads to saturation of the brighter ones, and might further result in amplification of high-frequency background noises. An adaptive illumination strategy to real-time-compress the dynamic range demands a dedicated hardware to operate and owing to electronic limitations, might encounter a poor effective bandwidth especially when each digitized pixel is required to be illumination optimized. Furthermore, such a method is often not immune to noise-amplification while locally enhancing a weak-intensity structure. We report a dedicated-hardware-free method for rapid noise-suppressed wide-dynamic-range compression so as to enhance visibility of such weak-intensity structures in terms of both contrast-ratio and signal-to-noise ratio while minimizing saturation of the brightest ones. With large-FOV aliasing-free two-photon fluorescence neuronal imaging, we validate its effectiveness by retrieving weak-intensity ultrafine structures amidst a strong noisy background. With compute-unified-device-architecture (CUDA)-acceleration, a time-complexity of <3 ms for a 1000x1000-sized 16-bit data-set is secured, enabling a real-time applicability of the same.

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Large field-of-view phase and fluorescence mesoscope with microscopic resolution

Cited by 13 publications

References 44 publications

Single-shot wavelength-multiplexed phase microscopy under Gabor regime in a regular microscope embodiment

Single-shot wavelength-multiplexed phase microscopy under Gabor regime in a regular microscope embodiment

A rapid denoised contrast enhancement method digitally mimicking an adaptive illumination in submicron-resolution neuronal imaging

Real-time Noise-suppressed Wide-Dynamic-Range Compression in Ultrahigh-Resolution Neuronal Imaging

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