Biological Imaging Applications Research Articles

Construction of a novel isopinocamphone-based fluorescent probe for rapid and specific detection of ClO− and its application in environmental analysis and biological imaging

A “turn on” fluorescent probe DMTI was designed for the detection of ClO− based on isopinocamphone.

The Role of Terminal and Bridge Ligands on the Molecular Upconversion of Lanthanide(III) 1D Coordination Polymers

The upconversion of trivalent lanthanide ions (LnIII) has gained significant attention in recent decades due to its potential applications in energy conversion and biological imaging. Recently, the concept of LnIII...

In vivo biological imaging applications of Bessel two-photon light-sheet microscopy

In vivo biological imaging applications of Bessel two-photon light-sheet microscopy

Tunable achromatic metalens for generating focused vortex beam in the near-infrared range

Highly precise and controllable focusing of optical vortex beams in the NIR range is essential for applications in biological imaging, nanomanipulation, and other fields. However, achieving tunable vortex beams across a broad spectrum remains a significant challenge. Herein, we propose a varifocal and broadband achromatic metalens capable of effectively correcting chromatic aberration, achieving a maximum focusing efficiency of 40.0% over 1400–1700 nm. Furthermore, through adjustments to the crystalline fraction of Ge2Sb2Te5 (GST), it offers the ability to vary focal lengths from 6.90 to 10.73 μm. This study may further advance NIR communication and imaging systems.

Fourier ptychographic microscopy image stack reconstruction using implicit neural representations

Image stacks provide invaluable 3D information in various biological and pathological imaging applications. Fourier ptychographic microscopy (FPM) enables reconstructing high-resolution, wide field-of-view image stacks without z-stack scanning, thus significantly accelerating image acquisition. However, existing FPM methods take tens of minutes to reconstruct and gigabytes of memory to store a high-resolution volumetric scene, impeding fast gigapixel-scale remote digital pathology. While deep learning approaches have been explored to address this challenge, existing methods poorly generalize to novel datasets and can produce unreliable hallucinations. This work presents FPM-INR, a compact and efficient framework that integrates physics-based optical models with implicit neural representations (INRs) to represent and reconstruct FPM image stacks. FPM-INR is agnostic to system design or sample types and does not require external training data. In our experiments, FPM-INR substantially outperforms traditional FPM algorithms with up to a 25-fold increase in speed and an 80-fold reduction in memory usage for continuous image stack representations.

Novel imidazo[1,2,4] triazole derivatives: Synthesis, fluorescence, bioactivity for SHP1

The Src homology 2 domain-containing protein tyrosine phosphatase 1 (SHP1) is a convergent node for oncogenic cell-signaling cascades. Consequently, SHP1 represents a potential target for drug development in cancer treatment. The development of efficient methods for rapidly tracing and modulating the SHP1 activity in complex biological systems is of considerable significance for advancing the integration of diagnosis and treatment of the related disease. Thus, we designed and synthesized a series of imidazo[1,2,4] triazole derivatives containing salicylic acid to explore novel scaffolds with inhibitory activities and good fluorescence properties for SHP1. The photophysical properties and inhibitory activities of these imidazo[1,2,4] triazole derivatives (5a-5y) against SHP1PTP were thoroughly studied from the theoretical simulation and experimental application aspects. The representative compound 5p exhibited remarkable fluorescence response (P: 0.002) with fluorescence quantum yield (QY) of 0.37 and inhibitory rate of 85.21 ± 5.17% against SHP1PTP at the concentration of 100 μM. Furthermore, compound 5p showed obvious aggregation caused quenching (ACQ) effect and had high selectivity for Fe3+ ions, good anti-interference and relatively low detection limit (5.55 μM). Finally, the cellular imaging test of compound 5p also exhibited good biocompatibility and certain potential biological imaging application. This study provides a potential way to develop molecules with fluorescent properties and bioactivities for SHP1.

Exchangeable Self-Assembled Lanthanide Antennas for PLIM Microscopy.

Lanthanides have unique photoluminescence (PL) emission properties, including very long PL lifetimes. This makes them ideal for biological imaging applications, especially using PL lifetime imaging microscopy (PLIM). PLIM is an inherently multidimensional technique with exceptional advantages for quantitative biological imaging. Unfortunately, due to the required prolonged acquisitions times, photobleaching of lanthanide PL emission currently constitutes one of the main drawbacks of PLIM. In this study, we report a small aqueous-soluble, lanthanide antenna, 8-methoxy-2-oxo-1,2,4,5-tetrahydrocyclopenta[de]quinoline-3-phosphonic acid, PAnt, specifically designed to dynamically interact with lanthanide ions, serving as exchangeable dye aimed at mitigating photobleaching in PLIM microscopy in cellulo. Thus, self-assembled lanthanide complexes that may be photobleached during image acquisition are continuously replenished by intact lanthanide antennas from a large reservoir. Remarkably, our self-assembled lanthanide complex clearly demonstrated a significant reduction of PL photobleaching when compared to well-established lanthanide cryptates, used for bioimaging. This concept of exchangeable lanthanide antennas opens new possibilities for quantitative PLIM bioimaging.

Exchangeable Self‐Assembled Lanthanide Antennas for PLIM Microscopy

AbstractLanthanides have unique photoluminescence (PL) emission properties, including very long PL lifetimes. This makes them ideal for biological imaging applications, especially using PL lifetime imaging microscopy (PLIM). PLIM is an inherently multidimensional technique with exceptional advantages for quantitative biological imaging. Unfortunately, due to the required prolonged acquisitions times, photobleaching of lanthanide PL emission currently constitutes one of the main drawbacks of PLIM. In this study, we report a small aqueous‐soluble, lanthanide antenna, 8‐methoxy‐2‐oxo‐1,2,4,5‐tetrahydrocyclopenta[de]quinoline‐3‐phosphonic acid, PAnt, specifically designed to dynamically interact with lanthanide ions, serving as exchangeable dye aimed at mitigating photobleaching in PLIM microscopy in cellulo. Thus, self‐assembled lanthanide complexes that may be photobleached during image acquisition are continuously replenished by intact lanthanide antennas from a large reservoir. Remarkably, our self‐assembled lanthanide complex clearly demonstrated a significant reduction of PL photobleaching when compared to well‐established lanthanide cryptates, used for bioimaging. This concept of exchangeable lanthanide antennas opens new possibilities for quantitative PLIM bioimaging.

Metal-free click and bioorthogonal reactions of aggregation-induced emission probes for lighting up living systems.

In 2002, two transformative research paradigms emerged: 'click chemistry' and 'aggregation-induced emission (AIE),' both leaving significant impacts on early 21st-century academia. Click chemistry, which describes the straightforward and reliable reactions for linking two building blocks, has simplified complex molecular syntheses and functionalization, propelling advancements in polymer, material, and life science. In particular, nontoxic, metal-free click reactions involving abiotic functional groups have matured into bioorthogonal reactions. These are organic ligations capable of selective and efficient operations even in congested living systems, therefore enabling in vitro to in vivo biomolecular labelling. Concurrently, AIE, a fluorogenic phenomenon of twisted π-conjugated compounds upon aggregation, has offered profound insight into solid-state photophysics and promoted the creation of aggregate materials. The inherent fluorogenicity and aggregate-emission properties of AIE luminogens have found extensive application in biological imaging, characterized by their high-contrast and photostable fluorescent signals. As such, the convergence of these two domains to yield efficient labelling with excellent fluorescence images is an anticipated progression in recent life science research. In this review, we intend to showcase the synergetic applications of AIE probes and metal-free click or bioorthogonal reactions, highlighting both the achievements and the unexplored avenues in this promising field.

Functional chromophores synthesized via multicomponent Reactions: A review on their use as cell-imaging probes

This review aims to provide a comprehensive overview of recent advancements and applications of fluorescence imaging probes synthesized via MCRs (multicomponent reactions). These probes, also known as functional chromophores, belong to a currently investigated class of fluorophores that are presently being successfully applied in bioimaging experiments, especially in various living cell lineages. We describe some of the MCRs that have been employed in the synthesis of these probes and explore their applications in biological imaging, with an emphasis on cellular imaging. The review also discusses the challenges and future perspectives in the field, particularly considering the potential impact of MCR-based fluorescence imaging probes on advancing this field of research in the coming years. Considering that this area of research is relatively new and nearly a decade has passed since the first publication, this review also provides a historical perspective on this class of fluorophores, highlighting the pioneering works published between 2011 and 2016.

Receptor for advanced glycation end-products: Biological significance and imaging applications.

The receptor for advanced glycation end-products (RAGE or AGER) is a transmembrane, immunoglobulin-like receptor that, due to its multiple isoform structures, binds to a diverse range of endo- and exogenous ligands. RAGE activation caused by the ligand binding initiates a cascade of complex pathways associated with producing free radicals, such as reactive nitric oxide and oxygen species, cell proliferation, and immunoinflammatory processes. The involvement of RAGE in the pathogenesis of disorders such as diabetes, inflammation, tumor progression, and endothelial dysfunction is dictated by the accumulation of advanced glycation end-products (AGEs) at pathologic states leading to sustained RAGE upregulation. The involvement of RAGE and its ligands in numerous pathologies and diseases makes RAGE an interesting target for therapy focused on the modulation of both RAGE expression or activation and the production or exogenous administration of AGEs. Despite the known role that the RAGE/AGE axis plays in multiple disease states, there remains an urgent need to develop noninvasive, molecular imaging approaches that can accurately quantify RAGE levels in vivo that will aid in the validation of RAGE and its ligands as biomarkers and therapeutic targets. This article is categorized under: Diagnostic Tools > In Vivo Nanodiagnostics and Imaging Diagnostic Tools > Biosensing.

A versatile AIE probe with two cross-talk-free emissions for dual detection of SO2 and viscosity and its application in food and biological imaging

Sulfur dioxide (SO2) and viscosity play important physiological roles in organisms, and real-time detection of their content changes is critical for organisms and human health. Herein, we developed a fluorescence probe NPNB with aggregation-induced emission (AIE) performance and mitochondrial localization effect for simultaneous detection of viscosity and SO2. NPNB can realize the efficient and specific detection of SO2 and viscosity at different emission wavelengths with a cross-talk-free effect, respectively. NPNB showed good sensing performance against SO2, including fast response (<7 min), and low detection limit (8.4 nM). Bioimaging experiments indicated that NPNB has good mitochondrial targeting and can achieve cellular imaging of both exogenous and endogenous SO2 and viscosity. Importantly, NPNB enables real-time imaging of changes in SO2 and viscosity during mitochondrial dysfunction. Moreover, the resulting probe has been applied to detect SO2 in food and water samples with a satisfactory recovery (84.46–105.57 %), further demonstrating its compatibility and practicality.

Single-shot wide-field full-stokes polarization imaging

Polarization is an intrinsic property of light. With the advancements in nanophotonics, it is now possible to achieve arbitrary polarization transformations in space. Incorporating special functions in different spatial dimensions expands the design freedom of metasurfaces and other optical devices. To understand and utilize the metasurface, we design a metasurface optical element to achieve single-shot, wide-field polarization imaging. Our demonstration exhibits a compact, full Stokes wide-field polarization imaging device without requiring conventional optical elements, such as moving parts or wide-field lens sets. Wide-field polarization imaging devices are potential candidates for machine vision and biological tissue imaging applications.

Gd3+-sensitized rare earth fluoride scintillators for High-resolution flexible X-ray imaging

X-ray imaging technology is widely used in the medical, industrial, and scientific fields, where highly sensitive, flexible, and cost-effective scintillation screens are crucial prerequisites. In this work, YF3:Gd3+/Tb3+ microparticles (MPs) were prepared via an ultrafast chemical precipitation method and exhibited excellent luminescent properties under the excitation of ultraviolet light and X-ray. It was found that Gd3+ doping could effectively enhance the photoluminescence (PL) and X-ray activated radioluminescence (RL) of Tb3+ owing to an efficient energy transfer process from Gd3+ to Tb3+. The prepared YF3:30%Gd3+/10%Tb3+ MPs exhibited fine environmental stability and superb X-ray RL, which was stronger than that of commercial scintillators Bi4Ge3O12 (BGO) and even comparable to that of CsI:TI single crystal. Highly flexible transparent YF3:Gd3+/Tb3+/PDMS scintillation screens were prepared to record high-resolution X-ray images of planar/nonplanar objects, where the imaging spatial resolution was 16.8 lp mm−1, which was far higher than the standard spatial resolutions required for medical computed tomography (CT) scan (2 lp mm−1) and X-ray imaging (3.5 lp mm−1). The excellent scintillation performance, superior flexibility and environmental stability of YF3:Gd3+/Tb3+/PDMS scintillation films endow them with the potential application in high-resolution industrial nondestructive inspection and biological imaging.

Enhancing quantum efficiency of Mn4+-doped fluoride phosphors through reverse doping strategy toward potential biological detection

Mn4+-doped fluoride red-emitting materials are renowned for their distinct characteristics, including fixed emission peaks and narrow spectral emission, rendering them highly favored in photoluminescent light-emitting diodes (LEDs). However, achieving ultra-high-concentration Mn4+ doping in fluoride phosphors is imperative to enhance quantum efficiency. Nevertheless, attaining such doping levels in metastable compound systems poses considerable challenges. In this study, we introduce an innovative reverse doping strategy, leading to the synthesis of Mn4+-doped cubic K2SiF6 phosphor (11.38 mol %) from hexagonal K2MnF6. Remarkably, this approach significantly improves the external quantum efficiency (EQE) to 66.7 % upon 450 nm excitation. Through systematic comparisons with conventional co-precipitation and ion exchange methods, our findings unequivocally demonstrate the superior efficacy of this approach in achieving high-concentration Mn4+ ion doping within relatively insoluble matrices. The successful application of our optimized phosphor enabled the fabrication of a red light-emitting diode (LED) that exhibits an exceptional photoelectric efficiency of 31.67 % at 350 mA, surpassing the performance of a commercial red LED chip. Furthermore, the stable red-light emission from the phosphor-converted LED (pc-LED) proves instrumental in rapid-response vascular imaging, showcasing the potential of our findings in advancing light sources for biological detection applications. This research provides valuable insights into enhancing red emission efficiency in slightly-soluble fluoride phosphors via novel synthetic strategy, unlocking possibilities for the development of highly efficient light sources in biological detection and imaging applications.

A spatially uniform illumination source for widefield multi-spectral optical microscopy.

Illumination uniformity is a critical parameter for excitation and data extraction quality in widefield biological imaging applications. However, typical imaging systems suffer from spatial and spectral non-uniformity due to non-ideal optical elements, thus require complex solutions for illumination corrections. We present Effective Uniform Color-Light Integration Device (EUCLID), a simple and cost-effective illumination source for uniformity corrections. EUCLID employs a diffuse-reflective, adjustable hollow cavity that allows for uniform mixing of light from discrete light sources and modifies the source field distribution to compensate for spatial non-uniformity introduced by optical components in the imaging system. In this study, we characterize the light coupling efficiency of the proposed design and compare the uniformity performance with the conventional method. EUCLID demonstrates a remarkable illumination improvement for multi-spectral imaging in both Nelsonian and Koehler alignment with a maximum spatial deviation of ≈ 1% across a wide field-of-view.

Light field measurement of specular surfaces by multi-polarization and hybrid modulated illumination.

Specular highlights present a challenge in light field microscopy imaging fields, leading to loss of target information and incorrect observation results. Existing highlight elimination methods suffer from computational complexity, false information and applicability. To address these issues, an adaptive multi-polarization illumination scheme is proposed to effectively eliminate highlight reflections and ensure uniform illumination without complex optical setup or mechanical rotation. Using a multi-polarized light source with hybrid modulated illumination, the system achieved combined multi-polarized illumination and physical elimination of specular highlights. This was achieved by exploiting the different light contributions at different polarization angles and by using optimal solution algorithms and precise electronic control. Experimental results show that the proposed adaptive illumination system can efficiently compute control parameters and precisely adjust the light source output in real time, resulting in a significant reduction of specular highlight pixels to less than 0.001% of the original image. In addition, the system ensures uniform illumination of the target area under different illumination configurations, further improving the overall image quality. This study presents a multi-polarization-based adaptive de-highlighting system with potential applications in miniaturization, biological imaging and materials analysis.

Construction of novel unsymmetrical rhodamine analogue and its application as nitric oxide probe

Traditional rhodamine dyes' absorption and emission wavelengths are in the visible region and Stokes shifts are less than 30 nm, which limit their biological imaging applications. To regulate the photophysical properties of xanthene chromophores, we reported a novel mitochondria-targeted unsymmetrical fluorescent thiophene-hybridized xanthene dye DTPO with deep-red emission wavelength (640 nm) and improved Stokes shift (58 nm) in water. DTPO exhibited multiple excellent properties, such as high molar extinction coefficients, large fluorescence quantum yields, tolerance to photobleaching. Meanwhile, the dye is lower toxic to living cell, and could accumulate in mitochondria. By using DTPO as a fluorophore, a highly selective fluorescent probe DTPO-OPD for the fast detection of mitochondrial NO was synthesized based on the NO-triggered the spiro-ring opening mechanism.

Poly(tetrasubstituted-aryl imidazole)s: A Way to Obtain Multi-Chromophore Materials with a Tunable Absorption/Emission Wavelength

Some original poly(tetrasubstituted imidazole)s incorporating different units were synthesized and characterized. These materials were obtained via a cascade polycondensation process assisted by microwave irradiation that was developed by our team. This time, we integrated two well-known chromophore structures into the macromolecular backbone, which were benzothiadiazole (BTD) and diketopyrrolopyrrole (DKPP). These new polymers were fully characterized: their chemical structures were confirmed using NMR spectroscopy and their thermal, optical and electrochemical properties were investigated and compared with a reference polymer containing a phenyl spacer instead of the mentioned chromophore units. These materials were found to exhibit a large Stokes shift of up to 350 nm. Furthermore, a polymer presenting large absorption on the UV–visible range and an emission close to the near-infrared region was obtained by coupling the mentioned moieties. According to the established properties of this latter polymer, it presents a potential for applications in biological imaging or optoelectronic devices.

Controlling ESIPT-based AIE effects for designing optical materials with single-component white-light emission

Aggregation-induced emission (AIE) molecules have gained significant importance in various fields such as biological imaging and organic light-emitting diodes. Here, we focused on exploring the process of the induction and control of AIE effects by modifying intramolecular hydrogen bonds. Firstly, inspired by the AIE characteristics of our reported amino-type excited-state intramolecular proton transfer (ESIPT)-based molecules, qualitative and quantitative relationships between the acidity of the amino group and the ESIPT process were investigated. They were successfully revealed through artificial intelligence modeling and quantum chemical calculations. Based on this, eight compounds, (2-(2′-aminophenyl) benzothiazole (1) and its derivatives), were synthesized. Then, their mechanism of structural and optical phenomena (AIE or aggregation-caused quenching (ACQ)) was further verified by quantum chemical calculations and experiments. It has been proved that multistage photochemical reactions could lead to the AIE phenomenon. Finally, the eight compounds were applied to biological imaging and white light material applications, benefiting from tunable dual-emission spectroscopic properties of amino compounds with ESIPT properties. This study provided extensive guidance on developing molecules with AIE properties and opened up new avenues for AIE-based principles, as well as single-component white-light emission optical materials.