Exosomes are small extracellular vesicles with diameters of 30-150 nm. In both physiological and pathological conditions, nearly all types of cells can release exosomes, which play important roles in cell communication and epigenetic regulation by transporting crucial protein and genetic materials such as miRNA, mRNA, and DNA. Consequently, exosome-based disease diagnosis and therapeutic methods have been intensively investigated. However, as in any natural science field, the in-depth investigation of exosomes relies heavily on technological advances. Historically, the two main technical hindrances that have restricted the basic and applied researches of exosomes include, first, how to simplify the extraction and improve the yield of exosomes and, second, how to effectively distinguish exosomes from other extracellular vesicles, especially functional microvesicles. Over the past few decades, although a standardized exosome isolation method has still not become available, a number of techniques have been established through exploration of the biochemical and physicochemical features of exosomes. In this work, by comprehensively analyzing the progresses in exosome separation strategies, we provide a panoramic view of current exosome isolation techniques, providing perspectives toward the development of novel approaches for high-efficient exosome isolation from various types of biological matrices. In addition, from the perspective of exosome-based diagnosis and therapeutics, we emphasize the issue of quantitative exosome and microvesicle separation.

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Progress, opportunity, and perspective on exosome isolation - efforts for efficient exosome-based theranostics.

This paper describes the main protocols that are used for fabricating microfluidic devices from glass and silicon. Methods for micropatterning glass and silicon are surveyed, and their limitations are discussed. Bonding methods that can be used for joining these materials are summarized and key process parameters are indicated. The paper also outlines techniques for forming electrical connections between microfluidic devices and external circuits. A framework is proposed for the synthesis of a complete glass/silicon device fabrication flow.

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A practical guide for the fabrication of microfluidic devices using glass and silicon

Advances in transdermal insulin delivery.

Single-objective selective-plane illumination microscopy (soSPIM) is achieved with micromirrored cavities combined with a laser beam-steering unit installed on a standard inverted microscope. The illumination and detection are done through the same objective. soSPIM can be used with standard sample preparations and features high background rejection and efficient photon collection, allowing for 3D single-molecule-based super-resolution imaging of whole cells or cell aggregates. Using larger mirrors enabled us to broaden the capabilities of our system to image Drosophila embryos.

3D high- and super-resolution imaging using single-objective SPIM

Metallization and nanostructuring of semiconductor surfaces by galvanic displacement processes

Etching characteristics and properties of {1 1 0} silicon crystal planes used as 45° optical mirrors for deflecting optical beams from/to optical fibers were investigated. Fiber aligning grooves and passive mirror-like planes were realized by wet micromachining of (1 0 0) silicon in KOH–IPA and TMAH–IPA systems. Implementation of Triton-x-100 surfactant as an additive to 25% TMAH in anisotropic etching of {1 1 0} silicon passive mirror planes is reported and discussed. It was found that Triton-x-100 contents in the range of 10–200 ppm to the 25% TMAH–water etchant significantly increase the anisotropy mostly by decreasing the {1 1 0} etch rate and retaining the {1 0 0} etch rate. It is also shown that {1 1 0} surface roughness is substantially improved compared to two other etching systems. Furthermore, efficient convex corner underetching reduction is demonstrated. The results of optical characterization of passive mirrors with 632 nm incident light show reduced scattering of reflected optical beam due to improved microroughness for mirrors made by TMAH–Triton. For the reflection of the optical beam with 1.33 µm and 1.54 µm wavelengths, sputtered layer of gold is used as reflective coating on silicon mirrors thus increasing the reflected optical beam intensity by an additional 8%.

http://iopscience.iop.org/article/10.1088/0960-1317/15/6/007/pdf

The role of Triton surfactant in anisotropic etching of {1 1 0} reflective planes on (1 0 0) silicon

Exosomes, a type of nanovesicle, are distinct cellular entities specifically capable of carrying various cargos between cells. It has been hypothesized that exosomes, as an enriched source of biomolecules, may serve as biomarkers for various diseases. This review introduces general aspects of exosomes, presents the challenges in exosome research, discusses the potential of exosomes as biomarkers, and describes the contribution of microfluidic technology to enable their isolation and analysis for diagnostic and disease monitoring. Additionally, clinical applications of exosomes for diagnostic purposes are also summarized.

https://www.mdpi.com/2072-666X/10/6/392/pdf

Microfluidic Technology for Clinical Applications of Exosomes.

Design, fabrication and characterization of thin film Ti/Pt heaters and integrated temperature sensors on a Si microfluidic platform are presented. Ti/Pt heaters and sensors provide controlled heating of microchannels realized on the opposite side of the Si platform. Ti/Pt heaters and sensors were fabricated simultaneously by a dc sputtering method and a lift-off process. Thermal annealing of deposited Ti/Pt layers in the temperature range of 300?700 ?C was investigated revealing a strong impact on the Ti/Pt resistivity and, consequently, on the final resistance of fabricated heaters and sensors. Furthermore, it was determined that the temperature coefficient of resistance (TCR) for Ti/Pt temperature sensors and the heater increased with the annealing temperature. Microstructural analysis of deposited and annealed Ti/Pt layers carried out by AES and AFM revealed that recrystallization followed by a grain growth process of heat-treated Ti/Pt layers started at around 500 ?C and correlated well with the behavior of electrical properties, but not with the TCR behavior of annealed layers. To reduce the heat losses of the heated Si platform, the heater and temperature sensors were covered hermetically by anodically bonded Pyrex glass with a prefabricated insulating cavity. According to this approach the power consumption was reduced by more than 25% due to the improved thermal insulation. Additional insulation steps implemented during thermal characterization of the assembled microfluidic platform further reduced the power consumption, but also increased the time response of the microfluidic reactor.

https://iopscience.iop.org/article/10.1088/0960-1317/21/2/025025/pdf

Experimental study of heat-treated thin film Ti/Pt heater and temperature sensor properties on a Si microfluidic platform

An experimental study of in vivo insulin delivery through microinjection by using hollow silicon microneedle array is presented. A case study was carried out on a healthy human subject in vivo to determine the influence of delivery parameters on drug transfer efficiency. As a microinjection device, a hollow microneedle array (13 × 13 mm2) having 100 microneedles (220 µm high, 130 µm-outer diameter and 50 µm-inner diameter) was designed and fabricated using classical microfabrication techniques. The efficiency of the delivery process was first characterized using methylene blue and a saline solution. Based on these results, the transfer efficiency was found to be predominantly limited by the inability of viable epidermis to absorb and allow higher drug transport toward the capillary-rich region. Two types of fast-acting insulin were used to provide evidence of efficient delivery by hollow MNA to a human subject. By performing blood analyses, infusion of more-concentrated insulin (200 IU/mL, international units (IU)) exhibited similar blood glucose level drop (5–7%) compared to insulin of standard concentration (100 IU/mL), however, significant increase of serum insulin (40–50%) with respect to the preinfusion values was determined. This was additionally confirmed by a distinctive increase of insulin to C-peptide ratio as compared to preinfusion ratio. Moreover, we noticed that this route of administration mimics a multiple dose regimen, able to get a “steady state” for insulin plasma concentration.

In Vivo Experimental Study of Noninvasive Insulin Microinjection through Hollow Si Microneedle Array

Investigation was focused on the formation of {311} planes by wet anisotropic etching of (100) silicon and, in particular, on the characterization by means of surface roughness, etch rates and related convex and concave corner dynamic behaviour during maskless etching. KOH and TMAH water solutions were tested for their influence on previously mentioned parameters as well as the effect of isopropyl alcohol (IPA). It was found that convex corner undercutting is significantly reduced if {311} bounding planes are utilized instead of {111} bounding planes. For shallow structures a self-compensation can be obtained with KOH and when certain conditions are met, also with TMAH. The rounding of the concave corner that arises through prolonged etching is reported, which is particularly emphasized in KOH and less in TMAH etchant. Addition of IPA in maskless mode is experimentally investigated, showing minor influence on etching conditions and on reducing the undercut of convex corners. Etch rates and dimensional control of some microstructures are discussed and presented comparatively for different etching systems in a temperature range of 50-100 °C. By evaluation of surface quality with a surface profiler and SEM, it was found that the smoothest surface was achieved by etching in TMAH. The role of solution temperature in surface roughness was found to be of minor importance, as well as the stirring of the solution. It was determined that the IPA additive increases roughness when used with KOH, while with TMAH, the influence on roughness of the {311} planes is insignificant.

http://iopscience.iop.org/article/10.1088/0960-1317/10/3/319/pdf

Danilo Vrtačnik

Papers

The role of Triton surfactant in anisotropic etching of {1 1 0} reflective planes on (1 0 0) silicon

Microfluidic Technology for Clinical Applications of Exosomes.

Experimental study of heat-treated thin film Ti/Pt heater and temperature sensor properties on a Si microfluidic platform

In Vivo Experimental Study of Noninvasive Insulin Microinjection through Hollow Si Microneedle Array

Morphological study of {311} crystal planes anisotropically etched in (100) silicon: role of etchants and etching parameters