Kidney microscopy is a mainstay in studying the morphological structure, physiology and pathology of kidney tissues, as histology provides important results for a reliable diagnosis. A microscopy modality providing at same time high-resolution images and a wide field of view could be very useful for analyzing the whole architecture and the functioning of the renal tissue. Recently, Fourier Ptychography (FP) has been proofed to yield images of biology samples such as tissues and in vitro cells while providing high resolution and large field of view, thus making it a unique and attractive opportunity for histopathology. Moreover, FP offers tissue imaging with high contrast assuring visualization of small desirable features, although with a stain-free mode that avoids any chemical process in histopathology. Here we report an experimental measuring campaign for creating the first comprehensive and extensive collection of images of kidney tissues captured by this FP microscope. We show that FP microscopy unlocks a new opportunity for the physicians to observe and judge renal tissue slides through the novel FP quantitative phase-contrast microscopy. Phase-contrast images of kidney tissue are analyzed by comparing them with the corresponding renal images taken under a conventional bright-field microscope both for stained and unstained tissue samples of different thicknesses. In depth discussion on the advantages and limitations of this new stain-free microscopy modality is reported, showing its usefulness over the classical light microscopy and opening a potential route for using FP in clinical practice for histopathology of kidney.

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Beyond conventional microscopy: Observing kidney tissues by means of fourier ptychography

: Quantitative phase imaging (QPI) is an ideal method to non-invasively monitor cell populations and provide label-free imaging and analysis. QPI offers enhanced sample characteri-zation and cell counting compared to conventional label-free techniques. We demonstrate this in the current study through a comparison of cell counting data from digital phase contrast (DPC) imaging and from QPI using a system based on Fourier ptychographic microscopy (FPM). Our FPM system offers multi-well, parallel imaging and a QPI-specific cell segmentation method to establish automated and reliable cell counting. Three cell types were studied and FPM showed improvement in the ability to resolve fine details and thin cells, despite limitations of the FPM system incurred by imaging artifacts. Relative to manually counted fluorescence ground-truth, cell counting results after automated segmentation showed improved accuracy with QPI over DPC.

Cellular analysis using label-free parallel array microscopy with Fourier ptychography

: All-optical ultrasound (OpUS) has emerged as an imaging paradigm well-suited to minimally invasive imaging due to its ability to provide high resolution imaging from miniaturised fibre optic devices. Here, we report a fibre optic device capable of concurrent laser interstitial thermal therapy (LITT) and real-time in situ all-optical ultrasound imaging for lesion monitoring. The device comprised three optical fibres: one each for ultrasound transmission, reception and thermal therapy light delivery. This device had a total lateral dimension of < 1 mm and was integrated into a medical needle. Simultaneous LITT and monitoring were performed on ex vivo lamb kidney with lesion depth tracked using M-mode OpUS imaging. Using one set of laser energy parameters for LITT (5 W, 60 s), the lesion depth varied from 3.3 mm to 8.3 mm. In all cases, the full lesion depth could be visualised and measured with the OpUS images and there was a good statistical agreement with stereomicroscope images acquired after ablation (t = 1.36, p = 0.18). This work demonstrates the feasibility and potential of OpUS to guide LITT in tumour resection.

Miniaturised dual-modality all-optical ultrasound probe for laser interstitial thermal therapy (LITT) monitoring

: Experimental and theoretical studies of colloidal nanoparticles have primarily focused on accurate characterization and simulation of observable characteristics, such as resonant wavelength. In this paper, we tackle the optimal design of colloidal-nanoparticle ensembles: what is the largest possible per-volume optical cross-section, which designs might achieve them, and can such response be experimentally demonstrated? We combine theory and experiment to answer each of these questions. We derive general bounds on the maximum cross-sections per volume, and we apply an analytical antenna model to show that resonant nanorods should nearly achieve such bounds. We use a modified seed-mediated synthesis approach to synthesize ensembles of gold nanorods with small polydispersity, i.e., small variations in size and aspect ratio. Polydispersity is the key determinant of how closely such ensembles can approach their respective bounds yet is difficult to characterize experimentally without near-field measurements. We show that a certain “extinction metric,” connecting extinction cross-section per volume with the radiative efficiencies of the nanoparticles, offers a quantitative prediction of polydispersity via quantities that can be rapidly measured with far-field characterization tools. Our predictions apply generally across all plasmonic materials and offer a roadmap to the largest possible optical response of nanoparticle ensembles.

Towards maximum optical efficiency of ensembles of colloidal nanorods

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Cellular analysis using label-free parallel array microscopy with Fourier ptychography

Miniaturised dual-modality all-optical ultrasound probe for laser interstitial thermal therapy (LITT) monitoring

Towards maximum optical efficiency of ensembles of colloidal nanorods