Quantitative single?cell optical technologies

Cells are the fundamental building bricks for all life. Optical technologies have served as mainstream single-cell analysis, which may trace back to 1600s when Robert Hooke reported “cell” in his groundbreaking publication of Micrographia. In 1900s, novel such Raman spectroscopy, flow cytometry, and fluorescence microscopy emerge that widely open fields cell analysis [1-3]. It is worthy point out invention laser has a great impact on study, not only used microscopic scanning but also cytometry [3, 4]. The Nobel laureates chemistry 2014 super-resolution (SR) pushed imaging be far below optical diffraction limit. Closely watching those quantitative technologies, we track obtaining better resolution higher throughput while measuring single cells (Table 1). Flow measures physical or chemical properties fluidic stream. been biomedicine its capability offer high data cells. Dating first generation scattered lights measured [5]. then comes era multi-fluorescence labeling can up around 20 markers per [6]. With advancement sensor technology, microfluidic machine learning so on, recently developed label-free in-vivo shown applications capabilities [7-9]. To observe specific structure dynamical processes life activity within living more clearly, some SR methods were proposed early 21st century, including photoactivated localization microscopy, stochastic reconstruction stimulated emission depletion structured illumination (SIM) [10-13] These extend limits allowing observation delicate structures clathrin-coated pits, cytoskeleton actin microtubulin, forth. SIM requires fewer photons among various suitable live-cell imaging. Nevertheless, prone artifacts, affects identification quantification true structure. Moreover, with still strong excitation light generate phototoxicity. Hessian invented by optimizing path design hardware exploiting spatiotemporal continuity biological algorithm, reduces phototoxicity traditional an order magnitude [14]. Most above mentioned rely allows organelles their interactions. Here review Wang et al. summarizes fluorescent probes techniques visualizing biomolecules at cellular, subcellular molecular scale using Caenorhabditis elegans major model organism (Wang al., this issue, p. XXX). Due limit, microscopes weaker than 200 nm, hardly fine organelles, usually resolved electron (Zhou However, microscopies reach about 10 nm. microscopies, SIM, suffer from aberrations thick sample [15], leading serious deterioration. Zheng report development adaptive optics based deep learning, attempting solve problem (Zheng Unlike most resonance energy transfer (FRET) interactions between molecules proteins. Yin develop mTA-G method could simply accurately measure critical factors FRET (Yin Label-free attracted wide attention research [5, 8, 16, 17]. Compared conventional labeling, non-invasive, cost-less, easily performed without demand operation skills labeling. Su colleagues scattering pattern-specific convolution net cytometer (LSPS-net Cytometer) cervical integration 2D (Liu work, they spatially distributed onto two-dimensional (2D) sensor. information-rich patterns analyzed automatic feature extractions, accuracies 90% obtained classification terms normal cancerous samples subtyping lines. Huang investigated biochip long-term in-situ after on-chip capture isolation (Fu Motivated simultaneous culturing cells, chip compact incubator. was self-interference spectroscopy (QSS). Their system capable incubator 130 h. By adopting QSS technique, realized obtain refractive index distribution resolution. Metze applied lifetime unstained smears bone marrow (da Silva In-vivo vessels instead other fabricated tubes, thus advantages study biomedicine. Aiming cytometric circulating tumor (CTCs) detection clinics, Wei measurements fluorescently labeled deoxy-glucose (Weng 2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl) amino]-2-deoxy-d-glucose (2-NBDG) label CTCs blood vessels, detected non-invasively cytometry. They due intake glucose mice level 2-NBDG effectively in-vivo. Zhu radio-frequency ablation (RFA), shows dramatically increased during RFA (Zhu interest notice technique image reconstruction, having demonstrated special issue analysis. Integration microfluidics improved measurements. here multidisciplinary lead breakthroughs human health. Major Science Technology Innovation Project Shandong Province, Grant number: 2019JZZY011016; National Natural Foundation China (NSFC), 91859114; sponsor: ZR2018MH032. Xuantao Su: Funding acquisition; writing-original draft. Liangyi Chen: Writing-original