High-Throughput 3D Imaging Flow Cytometry of Suspended Adherent 3D Cell Cultures.

3D cell cultures are indispensable in recapitulating in vivo environments. Among the many 3D culture methods, culturing adherent cells on hydrogel beads to form spheroid-like structures is a powerful strategy for maintaining high cell viability and functions in the adherent states. However, high-throughput, scalable technologies for 3D imaging of individual cells cultured on the hydrogel scaffolds are lacking.

Label-free mid-infrared photothermal live-cell imaging beyond video rate.

Advancement in mid-infrared (MIR) technology has led to promising biomedical applications of MIR spectroscopy, such as liquid biopsy or breath diagnosis. On the contrary, MIR microscopy has been rarely used for live biological samples in an aqueous environment due to the lack of spatial resolution and the large water absorption background. Recently, mid-infrared photothermal (MIP) imaging has proven to be applicable to 2D and 3D single-cell imaging with high spatial resolution inherited from visible light.

Adaptive dynamic range shift (ADRIFT) quantitative phase imaging.

Quantitative phase imaging (QPI) with its high-contrast images of optical phase delay (OPD) maps is often used for label-free single-cell analysis. Contrary to other imaging methods, sensitivity improvement has not been intensively explored because conventional QPI is sensitive enough to observe the surface roughness of a substrate that restricts the minimum measurable OPD. However, emerging QPI techniques that utilize, for example, differential image analysis of consecutive temporal frames, such as mid-infrared photothermal QPI, mitigate the minimum OPD limit by decoupling the static OPD contribution and allow measurement of much smaller OPDs.

Sequentially timed all-optical mapping photography boosted by a branched 4f system with a slicing mirror.

We present sequentially timed all-optical mapping photography (STAMP) with a slicing mirror in a branched 4f system for an increased number of frames without sacrificing pixel resolution. The branched 4f system spectrally separates the laser light path into multiple paths by the slicing mirror placed in the Fourier plane. Fabricated by an ultra-precision end milling process, the slicing mirror has 18 mirror facets of differing mirror angles.

Label-free detection of cysts using a deep learning-enabled portable imaging flow cytometer.

We report a field-portable and cost-effective imaging flow cytometer that uses deep learning and holography to accurately detect Giardia lamblia cysts in water samples at a volumetric throughput of 100 mL h-1. This flow cytometer uses lens free color holographic imaging to capture and reconstruct phase and intensity images of microscopic objects in a continuously flowing sample, and automatically identifies Giardia lamblia cysts in real-time without the use of any labels or fluorophores. The imaging flow cytometer is housed in an environmentally-sealed enclosure with dimensions of 19 cm × 19 cm × 16 cm and weighs 1.

Quantitative phase imaging with molecular vibrational sensitivity.

Quantitative phase imaging (QPI) quantifies the sample-specific optical-phase-delay enabling objective studies of optically transparent specimens such as biological samples but lacks chemical sensitivity, limiting its application to a morphology-based diagnosis. We present wide-field molecular vibrational (MV) microscopy realized in the framework of QPI utilizing a mid-infrared (MIR) photothermal effect. Our technique provides MIR spectroscopic performance comparable to that of a conventional infrared spectrometer in the molecular fingerprint region of 1450-1640  cm and realizes wide-field molecular imaging of a silica-polystyrene bead mixture over a 100  μm×100  μm area at 1 frame per second with the spatial resolution of 430 nm and 2-3 orders of magnitude lower fluence of ∼10  pJ/μm compared to other high-speed label-free molecular imaging methods, reducing photodamages to the sample.

Molecular contrast on phase-contrast microscope.

An optical microscope enables image-based findings and diagnosis on microscopic targets, which is indispensable in many scientific, industrial and medical settings. A standard benchtop microscope platform, equipped with e.g.

A deep learning-enabled portable imaging flow cytometer for cost-effective, high-throughput, and label-free analysis of natural water samples.

We report a deep learning-enabled field-portable and cost-effective imaging flow cytometer that automatically captures phase-contrast color images of the contents of a continuously flowing water sample at a throughput of 100 mL/h. The device is based on partially coherent lens-free holographic microscopy and acquires the diffraction patterns of flowing micro-objects inside a microfluidic channel. These holographic diffraction patterns are reconstructed in real time using a deep learning-based phase-recovery and image-reconstruction method to produce a color image of each micro-object without the use of external labeling.

Edge sparsity criterion for robust holographic autofocusing.

Autofocusing is essential to digital holographic imaging. Previously used autofocusing criteria exhibit challenges when applied to, e.g.

Design for sequentially timed all-optical mapping photography with optimum temporal performance.

A recently developed ultrafast burst imaging method known as sequentially timed all-optical mapping photography (STAMP) [Nat. Photonics8, 695 (2014)10.1038/nphoton.