PDMS-based microfluidic device with multi-height structures fabricated by single-step photolithography using printed circuit board as masters

We have developed a method for fabricating microfluidic devices with multi-height structures using single step photolithography. The whole fabrication process is executed by conventional printed circuit board (PCB) technology without the need of having access to clean room facilities. Specifically designed "windows" and "rims" architectures were printed on films that were used as photomasks. Different levels of protruding features on the PCB master were produced by exposing a photomask followed by chemical wet etching. Poly(dimethylsiloxane) (PDMS) was then moulded against the positive relief master to generate microfluidic structures. In this report, we described the fabrication of a microfluidic device featured with a multi-height "sandbag" structure for particle entrapment and peripheral microchannels. Controlled immobilization of biological cells and immunocytochemcial staining assays were performed to demonstrate the applicability of the microfluidic device for cellular analysis. The integrity of the microdevice remained stable under applied pressure, indicating the robustness of the elastic PDMS structures for analytical operation. The simple microfabrication process requires only low-cost materials and minimal specialized equipment and can reproducibly produce mask lines of about 20 microm in width, which is sufficient for most microfluidic applications.     

Identification of a set of genes showing regionally enriched expression in the mouse brain

Background

We have recorded responses from single neurons in murine visual cortex to determine the effectiveness of the input from the two murine cone photoreceptor mechanisms and whether there is any unique selectivity for cone inputs at this higher region of the visual system that would support the possibility of colour vision in mice. Each eye was stimulated by diffuse light, either 370 (strong stimulus for the ultra-violet (UV) cone opsin) or 505 nm (exclusively stimulating the middle wavelength sensitive (M) cone opsin), obtained from light emitting diodes (LEDs) in the presence of a strong adapting light that suppressed the responses of rods.

Results

Single cells responded to these diffuse stimuli in all areas of striate cortex. Two types of responsive cells were encountered. One type (135/323 – 42%) had little to no spontaneous activity and responded at either the on and/or the off phase of the light stimulus with a few impulses often of relatively large amplitude. A second type (166/323 – 51%) had spontaneous activity and responded tonically to light stimuli with impulses often of small amplitude. Most of the cells responded similarly to both spectral stimuli. A few (18/323 – 6%) responded strongly or exclusively to one or the other spectral stimulus and rarely in a spectrally opponent manner.

Conclusion

Most cells in murine striate cortex receive excitatory inputs from both UV- and M-cones. A small fraction shows either strong selectivity for one or the other cone mechanism and occasionally cone opponent responses. Cells that could underlie chromatic contrast detection are present but extremely rare in murine striate cortex.     

Tuning up or down the UV-induced apoptosis in Chinese hamster ovary cells with cell cycle inhibitors

Exposure to UVC induces apoptosis in Chinese hamster ovary (CHO.K1) cells. While studying the underlying mechanism, we found that a variety of cell cycle inhibitors, including colcemid, hydroxyurea and mimosine, enhance the UV-induced apoptosis in these cells. Such enhancement was not dependent on the cell cycle progression nor was it related to the difference in UV sensitivity at different phases of the cell cycle. The expression of p21(waf1/cip1), a general cyclin-dependent kinase (CDK) inhibitor, was deficient in CHO.K1 cells. Ectopic overexpression of the human p21 markedly increased the survival rates of the UV-irradiated cells in the presence of colcemid. In addition, roscovitine, a small-molecule inhibitor of CDK, also inhibited the UV-induced apoptosis. These observations suggest that deregulation of CDK activity may be critical in the UV-induced apoptosis in CHO.K1 cells.     

Effect of substrate temperature on electrical and magnetic properties of epitaxial La1−x Pb x MnO3 films

Epitaxial La_1−x Pb _x MnO₃ (LPMO) thin films, grown on (100) SrTiO₃ substrates by laser ablation technique at different temperatures between 600 and 850°C, have been characterized for electrical and magnetic properties. The temperature dependence of resistivity showed that the metal-insulator transition temperature (T _MI) decreases with increasing substrate temperature, which has been attributed to decrease in Pb content in the filsm. The YBa₂Cu₃O _x /La_1−x MnO₃ heterostructures, exhibiting both superconductivity and ferromagnetism, have been fabricated.     

A Criterion for Finite Topological Determinacy of Map-Germs

Let f, g: (Rⁿ, 0) (R^p, 0) be two C map-germs. Then f and gare C⁰-equivalent if there exist homeomorphism-germs h and lof (Rⁿ, 0) and (R^p, 0) respectively such that g = l f h^–1.Let k be a positive integer. A germ f is k-C⁰-determined ifevery germ g with j^k g(0) = j^k f(0) is C⁰-equivalent to f. Moreover,we say that f is finitely topologically determined if f is k-C⁰-determinedfor some finite k. We prove a theorem giving a sufficient conditionfor a germ to be finitely topologically determined. We explainthis condition below. Let N and P be two C manifolds. Consider the jet bundle J^k(N,P) with fiber J^k(n, p). Let z in J^k(n, p) and let f be suchthat z = j^kf(0). Define Whether (f) < k depends only on z, not on f. We can thereforedefine the set Let W^k(N, P) be the subbundle of J^k(N, P) with fiber W^k(n, p).Mather has constructed a finite Whitney (b)-regular stratificationS^k(n, p) of J^k(n, p) – W^k(n, p) such that all strata aresemialgebraic and K-invariant, having the property that if S^k(N,P) denotes the corresponding stratification of J^k(N, P) –W^k(N, P) and f C(N, P) is a C map such that j^kf is multitransverseto S^k(N, P), j^kf(N) W^k(N, P) = and N is compact (or f is proper),then f is topologically stable. For a map-germ f: (Rⁿ, 0) (R^p, 0), we define a certain ojasiewiczinequality. The inequality implies that there exists a representativef: U R^p such that j^kf(U – 0) W^k (Rⁿ, R^p = and suchthat j^kf is multitransverse to S^k (Rⁿ, R^p) at any finite setof points S U – 0. Moreover, the inequality controlsthe rate j^kf becomes non-transverse as we approach 0. We showthat if f satisfies this inequality, then f is finitely topologicallydetermined. 1991 Mathematics Subject Classification: 58C27.     

Biofunctional organic nanocrystals for quantitative detection of pathogen deoxyribonucleic acid

Advances in nanotechnology have had significant impacts in the field of biodiagnostics. In this study, we describe the novel application of dissolvable, organic and biofunctional nanocrystals for the quantitative detection of a PCR product. Fluorescein diacetate (FDA), a fluorogenic precursor of fluorescein, was milled in a solution of a polymeric surfactant to create a stable, nanosized colloid with an interface for coupling streptavidin molecules. The application of these particulate labels for the quantitative detection of biotinylated human papillomavirus (HPV) DNA, amplified in a standard PCR procedure, was demonstrated. After the affinity reaction, the FDA molecules were dissolved and concomitantly converted into fluorescein. This approach resulted in a high selectivity, short incubation times and a sensitivity up to 147 times greater than obtained from state-of-the-art, directly fluorescent-labeled streptavidins. This innovative method offers rapid detection of small amounts of nucleic acids because less target material and thus fewer PCR cycles are required.