Springer Science and Business Media LLC

A portable, fully automated analyzer that provides actuation and flow control to a disposable, self-contained, microfluidic cassette (“chip”) for point-of-care, molecular testing is described. The analyzer provides mechanical actuation to compress pouches that pump liquids in the cassette, to open and close diaphragm valves for flow control, and to induce vibrations that enhance stirring. The analyzer also provides thermal actuation for the temperature cycling needed for polymerase chain reaction (PCR) amplification of nucleic acids and for various drying processes. To improve the temperature uniformity of the PCR chamber, the system utilizes a double-sided heating/cooling scheme with a custom feedforward, variable, structural proportional-integral-derivative (FVSPID) controller. The analyzer includes a programmable central processing unit that directs the sequence and timing of the various operations and that is interfaced with a computer. The disposable cassette receives a sample, and it carries out cell lysis, nucleic acid isolation, concentration, and purification, thermal cycling, and either real time or lateral flow (LF) based detection. The system’s operation was demonstrated by processing saliva samples spiked with B. cereus cells. The amplicons were detected with a lateral flow assay using upconverting phosphor reporter particles. This system is particularly suited for use in regions lacking centralized laboratory facilities and skilled personnel.

Density gradient centrifugation exploits density differences between different blood cells to accomplish separation of peripheral blood mononuclear cells (PBMCs) from polymorphonuclear (PNM) cells, and erythrocytes or red blood cells (RBCs). While density gradient centrifugation offers a label-free alternative avoiding the use of harsh lysis buffers for blood cell isolation, it is a time-consuming and labor-intensive process during which blood cells are subject to high-levels of centrifugal force that can artifactually activate cells. To provide a low-stress alternative to this elegant method, we miniaturized and automated this process using microfluidics to ensure continuous PBMCs isolation from whole blood while avoiding the exposure to high-levels of centrifugal stress in a simple flow-through format. Within this device, a density gradient is established by exploiting laminar flow within microfluidic channels to layer a thin stream of blood over a larger stream of Ficoll. Using this approach we demonstrate successful isolation of PBMCs from whole blood with preservation of monocytes and different lymphocyte subpopulations similar to that seen with conventional density gradient centrifugation. Evaluation of activation status of PBMCs isolated using this technique shows that our approach achieves minimal isolation process induced activation of cells in comparison to conventional lysis or density gradient centrifugation. This simple, automated microfluidic density gradient centrifugation technique can potentially serve as tool for rapid and activation-free technique for isolation of PBMCs from whole blood for point-of-care applications.

The development of the micro/nano science and technology has promoted the evolvement of human civilization tremendously. The advancement of the micro/nano science and technology highly depends on the progress of the micro/nano manipulation techniques, and the micro/nano-scaled manipulation level is the critical sign of the micro/nano science and technology. This review, aimed at the demand and the challenge of the micro/nano material and biomedical fields and related to the scientific issues and implementation techniques of the optically induced di-electrophoresis (ODEP). We explained its working principle, manipulating method, and influencing factors of ODEP force to a certain extent. A number of application fields based-ODEP technology and specific applications so far are summarized and reviewed. Finally, some perspectives are provided on current development trends, future research directions, and challenges of ODEP.

Development of a compact fluorescence-based detection system for use in a micro-analytical system, such as a point-of-care diagnostic system, often requires a multi-channel microfluidic chip system. Since the materials used for microfluidic chips usually are transparent in the visible region and have a refractive indices higher than that of air or the surrounding environment, the fluorescence emission and scattered excitation light can propagate through the chip. We observed that such propagation can cause cross-talk between adjacent channels, and may become the major source of noise in the system and/or photobleach the fluorescent samples in the adjacent channels, particularly for the small distances between the channels found in microfluidic chips, usually in order of several μ m. We monitored this cross-talk using fluorescein as a fluorescent sample and Mylar sheeting as a microfluidic chip material. We then discuss how this cross-talk can be avoided using a simple, inexpensive and effective method.

A micromachined drop dispenser for biofluidic applications has been fabricated and tested. It uses anisotropic etching of silicon and anodic bonding to pyrex wafers to form the flow channels, fluid wells, and nozzles. The ejected fluid comes from a fluid well that is externally compressed by a PZT bimorph that compresses a thin silicon membrane wall on the driven fluid well. This device design is amenable to arrays of dispensers although only one channel was fluid tested. It produced uniform droplets 150 µm in diameter.

Growth of human connective tissue progenitor cells (CTPs) was characterized on smooth and microtextured polydimethylsiloxane (PDMS) surfaces. Human bone marrow derived cells were cultured for nine days under conditions promoting osteoblastic differentiation on Smooth PDMS and PDMS Channel microtextures (11 μm high, 45 μm wide channels, and separated by 5 μm wide ridges). Glass tissue culture dish surfaces were used as controls. Cell numbers per colony, cell density within colonies, alignment of cells, area of colonies, and colony shapes were determined as a function of substrate surface topography. An alkaline phosphatase stain was used as a marker for osteoblastic phenotype. CTPs attached, proliferated, and differentiated on all surfaces with cell process lengths of up to 80 μm. Cells on the Smooth PDMS and control surfaces spread and proliferated as colonies in proximity to other cells and migrated in random directions creating colonies that covered significantly larger areas (0.96 and 1.05 mm2, respectively) than colonies formed on PDMS Channel textures (0.64 mm2). In contrast, cells on PDMS Channel textures spread, proliferated, aligned along the channel axis, and created colonies that were more dense, and with lengths of longest colony axes that were significantly longer (3252 μm) than those on the Smooth PDMS (1265 μm) and control surfaces (1319 μm). Cells on PDMS Channel textures were aligned at an angle of 14.44° relative to the channel axis, and the resulting colonies exhibited a significantly higher aspect ratio (13.72) compared to Smooth PDMS (1.57) and control surfaces (1.51).

Developed within the last few decades, microneedles (MNs) have only recently seen wide-scale use among the general population, especially in the area of cosmetics. With the FDA only starting to regulate microneedling devices and the many new microneedling products that enter the modern global market, it is of utmost importance to establish the safety profile and reasonable expectations of the microneedling practice and its products. In our review of current literature, the authors searched the keyword “microneedle” with the following terms: “safety”, “side effect”, “toxicology”, “adverse effect”, “adverse event”, “infection”, “dermatitis”, “granuloma”, “scarring”, and “hyperpigmentation”. Despite wide-scale implementation of MNs, we are likely only beginning to understand the potential of MNs as a medical and consumer product, and we should, therefore, be aware of any potential adverse events associated with the product.

The standard of care for posterior segment disorders such as wet age-related macular degeneration, diabetic macular oedema and retinal vascular occlusions is pharmacotherapy by intravitreal drug delivery. Since the therapeutic effect of these drugs lasts only around 4 to 8 weeks, repeated intravitreal injections are required. Pain is experienced by the patients during injection as the needle courses through the sclera and choroid. The current work describes the design and development of a novel anodized titanium alloy implant that allows for intravitreal injections through the implant so that the needle transverses only the conjunctiva, thus minimizing discomfort to the patient. Both ex-vivo testing of the implant in enucleated goat’s eye as well as in-vivo validation in rabbit eyes was carried out. The implant was placed through pars plana via a minor surgical procedure and was sutured to the sclera and covered with conjunctiva. Subsequent intravitreal injections were administered under topical anaesthesia with a 30-gauge needle through the implant thus delivering the drug into the vitreous cavity. Repeated intravitreal injections were administered every 2 weeks via the implant for 3 months in 4 rabbits. Apart from cataract in 1 rabbit, no complications were observed. There was no evidence of intra-ocular inflammation or infection at final follow-up. Histopathological analysis did not reveal any inflammation or necrosis around the area of implant. The implants were subsequently removed at 5 months and scleral wound was closed with a single suture. The sclera and overlying conjunctiva healed well and no intraocular complications were observed after removal.

Surgeons typically rely on their past training and experiences as well as visual aids from medical imaging techniques such as magnetic resonance imaging (MRI) or computed tomography (CT) for the planning of surgical processes. Often, due to the anatomical complexity of the surgery site, two dimensional or virtual images are not sufficient to successfully convey the structural details. For such scenarios, a 3D printed model of the patient’s anatomy enables personalized preoperative planning. This paper reviews critical aspects of 3D printing for preoperative planning and surgical training, starting with an overview of the process-flow and 3D printing techniques, followed by their applications spanning across multiple organ systems in the human body. State of the art in these technologies are described along with a discussion of current limitations and future opportunities.