RESUMO
This study presents a novel label-free approach for characterizing cell death states, eliminating the need for complex molecular labeling that may yield artificial or ambiguous results due to technical limitations in microscope resolution. The proposed holographic tomography technique offers a label-free avenue for capturing precise three-dimensional (3D) refractive index morphologies of cells and directly analyzing cellular parameters like area, height, volume, and nucleus/cytoplasm ratio within the 3D cellular model. We showcase holographic tomography results illustrating various cell death types and elucidate distinctive refractive index correlations with specific cell morphologies complemented by biochemical assays to verify cell death states. These findings hold promise for advancing in situ single cell state identification and diagnosis applications.
Assuntos
Morte Celular , Holografia , Imageamento Tridimensional , Tomografia , Holografia/métodos , Tomografia/métodos , Imageamento Tridimensional/métodos , Humanos , Refratometria/métodosRESUMO
Data acquisition and processing is a critical issue for high-speed applications, especially in three-dimensional live cell imaging and analysis. This paper focuses on sparse-data sample rotation tomographic reconstruction and analysis with several noise-reduction techniques. For the sample rotation experiments, a live Candida rugosa sample is used and controlled by holographic optical tweezers, and the transmitted complex wavefronts of the sample are recorded with digital holographic microscopy. Three different cases of sample rotation tomography were reconstructed for dense angle with a step rotation at every 2°, and for sparse angles with step rotation at every 5° and 10°. The three cases of tomographic reconstruction performance are analyzed with consideration for data processing using four noise-reduction techniques. The experimental results demonstrate potential capability in retaining the tomographic image quality, even at the sparse angle reconstructions, with the help of noise-reduction techniques.
Assuntos
Holografia/instrumentação , Holografia/métodos , Tomografia/instrumentação , Tomografia/métodos , Aprendizado Profundo , Processamento de Imagem Assistida por Computador , Imageamento Tridimensional , Pinças Ópticas , Rotação , Saccharomycetales , Razão Sinal-RuídoRESUMO
The noninvasive monitoring of protein secretion of cells responding to drug treatment is an effective and essential tool in latest drug development and for cytotoxicity assays. In this work, a surface functionalization method is demonstrated for specific detection of protein released from cells and a platform that integrates highly sensitive optical devices, called whispering-gallery mode biosensors, with precise microfluidics control to achieve label-free and real-time detection. Cell biomarker release is measured in real time and with nanomolar sensitivity. The surface functionalization method allows for antibodies to be immobilized on the surface for specific detection, while the microfluidics system enables detection in a continuous flow with a negligible compromise between sensitivity and flow control over stabilization and mixing. Cytochrome c detection is used to illustrate the merits of the system. Jurkat cells are treated with the toxin staurosporine to trigger cell apoptosis and cytochrome c released into the cell culture medium is monitored via the newly invented optical microfluidic platform.
Assuntos
Técnicas Biossensoriais , Microfluídica , Dispositivos Ópticos , Proteínas/metabolismo , Calibragem , Sobrevivência Celular , Citocromos c/metabolismo , Humanos , Células Jurkat , Proteínas Recombinantes/metabolismo , Propriedades de SuperfícieRESUMO
Transfer of liquid from one surface to another plays a vital role in printing processes. During liquid transfer, a liquid bridge is formed and subjected to substantial extension but incomplete liquid transfer can produce defects that are detrimental to the operation of printed electronic devices. One strategy for minimizing these defects is to apply an electric field, a technique known as electrostatic assist (ESA). However, the physical mechanisms underlying ESA remain a mystery. To better understand these mechanisms, slender-jet models are developed for both perfect dielectric and leaky dielectric axisymmetric Newtonian liquid bridges with moving contact lines. Nonlinear partial differential equations describing the evolution of the bridge radius and interfacial charge are derived and then solved using finite element methods. For perfect dielectrics, application of an electric field enhances liquid transfer to the more wettable surface over a wide range of capillary numbers. The electric field modifies the pressure differences inside the liquid bridge and, as a consequence, drives liquid toward the more wettable surface. For leaky dielectrics, charge can accumulate at the liquid-air interface. Application of an electric field can augment or oppose the influence of wettability differences, depending on the direction of the electric field and the sign of the surface charge. Flow visualization experiments reveal that when an electric field is applied, more liquid is transferred to the more wettable surface because of a modified bridge shape that causes depinning of the contact line. The measured values of the amount of liquid transferred are in good agreement with predictions of the perfect dielectric model.
RESUMO
Transfer of liquid from one surface to another plays a key role in printing processes. During liquid transfer, a liquid bridge is formed and then undergoes significant extensional motion while its contact lines are free to move on the bounding solid surfaces. In this work, we develop one-dimensional (1D) slender-jet and two-dimensional (2D) axisymmetric models of this phenomenon and compare the resulting predictions with previously published experimental data. For very low capillary numbers (Ca) (quasi-static stretching), predictions from both models of the amount of liquid transferred agree well with the experimental data, provided that the difference in receding contact angles (|Δθr|) between the two surfaces is sufficiently large. Notably, the amount of liquid transferred is primarily governed by the overall bridge shape and is not significantly influenced by contact-line motion toward the end of bridge stretching. For O(1) values of Ca, the models predict that each surface receives half the liquid, in agreement with experimental observations. For intermediate values of Ca (and very low values of Ca when |Δθr| is small enough), predictions from each model can deviate substantially from the experimental data, which we speculate is due to the influence of surface heterogeneities that are not included in the models. In this regime, there can be significant differences between the predictions of the 1D and 2D models, which is due to the tendency of the contact line to slip more in the 1D model. The models are also used to understand the influence of initial bridge shape on liquid transfer and to rationalize related experimental observations. The results from these fundamental studies will aid the optimization of gravure and other printing processes for manufacturing of printed electronic devices.
RESUMO
Rose petals exhibit superhydrophobicity with strong adhesion to pin water drops, known as the 'petal effect.' It is generally believed that the petal effect is attributed to dual-scale roughness, that is, the surface possesses both a nanostructure and a microstructure (Feng et al 2008 Langmuir 24 4114). In this study, we demonstrate that the dual-scale roughness is not a necessary condition for a surface of the petal effect. A surface of single-scale roughness, either at the nanoscale or the microscale alone, within a certain roughness region may also exhibit the petal effect. The surface roughness plays the essential role on the wetting behavior and governs the contact angle in the Wenzel or Cassie state, as well as the contact angle hysteresis. A water drop on the surface of the petal effect under the condition of the advancing and receding contact angle would fall into, respectively, the Cassie and Wenzel state, which leads to a contact angle hysteresis large enough to pin the water drop. On both single and dual textured hydrophobic surfaces, a sequence of wetting transitions: Wenzel state â petal state (sticky superhydrophobic state) â lotus state (slippery superhydrophobic state) is consistently observed by simply increasing the surface roughness.