Raman fingerprinting of leukemic cells has potential applications in diagnosis and in vitro chemosensitivity assessment. A biochemical map of the contents of leukemic cells can not only help distinguish cancer patients from healthy ones but also shed light on different subtypes of leukemia such as ALL, AML, etc. Certain important requirements need to be fulfilled to effectively measure the Raman map of a single leukemic cell. Firstly, since the leukemia cells are suspension cells, it is preferred to keep them in a free solution rather than attached to a fixed surface during signal acquisition. Secondly, the cells need to be immobilized for several seconds, for the acquisition of the weak Raman signal even when using stimulated Raman Spectroscopy (SRS) which provides relatively stronger Raman signal. Thus, a device capable of sequentially flowing, holding, and releasing individual leukemia cells in a robust, efficient and high-throughput manner is required. We present an optofluidic fiber tweezers device comprised of a novel combination of 3D hydrodynamic flow focusing and optical fiber in a microfluidic chip. By exploiting the interplay between the optical and hydrodynamic forces acting on the cell, we demonstrate rapid, efficient, sequential delivery and trapping of single leukemic cells in a flow cytometer format followed by SRS imaging of the trapped cell. The specific Raman vibration bands corresponding to the lipids, nucleic acids, and proteins in the trapped cells were analyzed to distinguish cancerous cells from healthy cells. Our device is also capable of isolating cells with unique Raman signatures for further processing using techniques like gene sequencing etc.
Detection and quantification of bacterial populations in droplets is a fundamental prerequisite for the application of droplet microfluidic technology in antibiotic susceptibility assays, paving the way for single-cell profiling and quantifying hetero-resistance. While several label-free detection approaches have been proposed, limited detection sensitivity and the ability to quantify bacterial populations in droplets accurately remain challenging. Furthermore, these approaches are prone to a high number of false positives resulting in low accuracy and requiring highly monodisperse droplets. This study presents a speckle image-based detection technique for the quantification of the bacterial population in droplets. Speckles are generated by scattering laser light from bacteria-loaded droplets in a flow cytometric approach. Spatial segregation of the scattering signal from the droplet surface as well as its contents allows the detection of encapsulated bacteria with a high signal-to-noise ratio. This results in a detection sensitivity of ⁓100 CFU/droplet, the highest achieved by any label-free detection technique in a flow format thus far. It also allows the identification of false-positive signals, thereby increasing the accuracy of detection and enabling operation with polydisperse droplets (diameter: 10–500 µm). The properties of the speckle image generated from droplets, such as speckle grain size and density, can be used to quantify the population of bacteria in droplets. This detection approach applies to a wide range of bacteria species of clinical and industrial importance, creating avenues for innovation in bacteria analysis using droplet microfluidics.
The flourishing field of light-powered micro/nanorotors provides promising strategies for manufacturing and biomedical needs. However, the torque of optical rotors typically arises from the momentum exchange with photons, which limits the geometries and materials of objects that can be rotated and requires intense laser beams with designed intensity profile and polarization. These factors inhibit the light-powered rotation of highly symmetric or isotropic targets. Herein, we developed an optothermal micro/nanorotors platform that enables the rotation of various colloids with diverse sizes, materials, and various shapes, including live cells and micro/nanoparticles with high symmetry and isotropy. The long-sought-after out-of-plane rotation has been achieved by a single plane-polarized Gaussian laser beam with an ultralow power. This simple rotor approach is foreseen to open new horizons in colloidal and life sciences by offering a non-invasive and universal manipulation.
We present opto-thermoelectric speckle tweezers (OTEST) for large-scale and high-throughput trapping of particles. OTEST combine optical speckle with plasmonic substrate to generate a thermal speckle field that consists of many random thermal hotspots to trap a large number of particles using thermoelectric forces. We demonstrate trapping of dielectric and metallic particles with sizes as small as 100 nm in the speckle field. Finally, we integrate OTEST with microfluidic systems to demonstrate filtration of the smaller-sized particles from a mixed solution of 200 nm and 1 µm particles.
Convection- and bubble-assisted nanoaperture-based plasmonic tweezers are presented to overcome the diffusion-limited trapping. Opto-thermally generated convection and bubble-induced flows rapidly transported particles from large spatial extent to plasmonic nanoapertures without relying on diffusion. The trapping time was reduced by more than order of magnitude. Moreover, the trapping time was brought within practical time limits at ultralow particle concentrations for which it could take several hours to trap a single particle.
Nanoaperture based trapping has developed as a significant tool among the various optical tweezer systems for trapping of very small particles down to the single nanometer range. The double nanohole aperture based trap provides a method for efficient, highly-sensitive, label-free, low-cost, free-solution single molecule trapping and detection. We use the double nanohole tweezer to understand biomolecular phenomena like protein unfolding, binding, structural conformation of DNA, protein-DNA interactions, and protein small molecule interactions.
The nanoplasmonic properties of apertures in metal films have been studied extensively; however, we have recently
discovered surprising new features of this simple system with applications to super-focusing and super-scattering.
Furthermore, apertures allow for optical tweezers that can hold onto particles of the order of 1 nm; I will briefly
highlight our work using these apertures to study protein - small molecule interactions and protein - DNA binding.
In this paper we describe the double nanohole laser tweezer system used to trap single nanoparticles. We cover the basic theory behind the DNH and what makes it more powerful than traditional laser tweezers commonly used for larger particles. We outline the basic setup used to reliably trap several different types of particles ranging in size from 1 nm to 40 nm. Data from several experiments is shown which displays exactly how a particle is confirmed to be trapped. We will discuss the use of autocorrelation as well as other information that can be extracted from the optical transmission in our setup and how it has been applied to the identification of protein small molecule interactions and protein binding. Other uses of the data collected from our setup will be discussed including the observation of protein folding. Finally we discuss the current developments of the process and its possible uses as a drug discovery tool, a new type of single particle nanopipette and new bio-sensors.
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