The extracellular matrix microtopography drives critical changes in cellular motility and Rho A activity in colon cancer cells.

We have shown that the microtopography (mT) underlying colon cancer changes as a tumor de-differentiates. We distinguish the well-differentiated mT based on the increasing number of "pits" and poorly differentiated mT on the basis of increasing number of "posts." We investigated Rho A as a mechanosensing protein using mT features derived from those observed in the ECM of colon cancer. We evaluated Rho A activity in less-tumorogenic (Caco-2 E) and more tumorigenic (SW620) colon cancer cell-lines on microfabricated pits and posts at 2.5 mum diameter and 200 nm depth/height. In Caco-2 E cells, we observed a decrease in Rho A activity as well as in the ratio of G/F actin on surfaces with either pits or posts but despite this low activity, knockdown of Rho A led to a significant decrease in confined motility suggesting that while Rho A activity is reduced on these surfaces it still plays an important role in controlling cellular response to barriers. In SW620 cells, we observed that Rho A activity was greatest in cells plated on a post microtopography which led to increased cell motility, and an increase in actin cytoskeletal turnover.

Nanofabrication in cellulose acetate.

We have demonstrated nanofabrication with commercialized cellulose acetate. Cellulose acetate is used for bulk nanofabrication and surface nanofabrication. In bulk nanofabrication, cellulose acetate reacts with an e-beam and permanent patterns are formed in it instead of being transferred to other substrates. We have studied the nano relief modulation performance of cellulose acetate before and after development. The depth of the nanopatterns is magnified after development, and is varied by exposing dosage and line width of the pattern. The thinnest 65 nm wide line is achieved in the bulk fabrication. We also demonstrate a binary phase Fresnel lens array which is directly patterned in a cellulose acetate sheet. Because of its unique mechanical and optical properties, cellulose is a good candidate for a template material for soft imprinting lithography. In the surface nanofabrication, cellulose acetate thin film spin-coated on silicon wafers is employed as a new resist for e-beam lithography. We achieved 50 nm lines with 100 nm pitches, dots 50 nm in diameter, and single lines with the smallest width of 20 nm. As a new resist of e-beam lithography, cellulose acetate has high resolution comparable with conventional resists, while having several advantages such as low cost, long stock time and less harmfulness to human health.

Nanosized corners for trapping and detecting magnetic nanoparticles.

We present a device concept based on controlled micromagnetic configurations in a corner-shaped permalloy nanostructure terminated with two circular disks, specifically designed for the capture and detection of a small number of magnetic beads in suspension. A transverse head-to-head domain wall (TDW) placed at the corner of the structure plays the role of an attracting pole for magnetic beads. The TDW is annihilated in the terminating disks by applying an appropriate magnetic field, whose value is affected by the presence of beads chemically bound to the surface. In the case where the beads are not chemically bound to the surface, the annihilation of the TDW causes their release into the suspension. The variation of the voltage drop across the corner, due to the anisotropic magnetoresistance (AMR) while sweeping the magnetic field, is used to detect the presence of a chemically bound bead. The device response has been characterized by using both synthetic antiferromagnetic nanoparticles (disks of 70 nm diameter and 20 nm height) and magnetic nanobeads, for different thicknesses of the protective capping layer. We demonstrate the detection down to a single nanoparticle, therefore the device holds the potential for the localization and detection of small numbers of molecules immobilized on the particle's functionalized surface.

Treatment with Y-27632, a ROCK Inhibitor, Increases the Proinvasive Nature of SW620 Cells on 3D Collagen Type 1 Matrix.

The concept of using tissue density as a mechanism to diagnose a tumor has been around for centuries. However, this concept has not been sufficiently explored in a laboratory setting. Therefore, in this paper, we observed the effects of cell density and extracellular matrix (ECM) density on colon cancer invasion and proliferation using SW620 cells. We also attempted to inhibit ROCK-I to determine its effect on cell invasion and proliferation using standard molecular biology techniques and advanced imaging. Increasing cell seeding density resulted in a 2-fold increase in cell invasion as well as cell proliferation independent of treatment with Y-27632. Increasing collagen I scaffold density resulted in a 2.5-fold increase in cell proliferation while treatment with Y-27632 attenuated this effect although 1.5 fold increase in cell invasion was observed in ROCK inhibited samples. Intriguingly, ROCK inhibition also resulted in a 3.5-fold increase in cell invasion within 3D collagen scaffolds for cells seeded at lower densities. We show in this paper that ROCK-I inhibition leads to increased invasion within 3D collagen I microenvironments. This data suggests that although ROCK inhibitors have been used clinically to treat several medical conditions, its effect largely depends on the surrounding microenvironment.

Micro/nanosized refractive lens arrays formed by means of conformal thin film deposition.

We provide a 'growing' method for fabricating a microlens array with lateral size of a few microns or less. Instead of using complicated etching techniques, the method forms a spherical profile of the lens using conformal chemical vapor deposition. We have fabricated two lens arrays. One has a pitch of 1200 nm, a circular aperture 1000 nm in diameter and a sag height of 130 nm. The other array has a pitch of 600 nm, and a square aperture of 600 nm × 600 nm, with a fill factor close to 100%. The maximum profile deviation between the fabricated lens and an ideal sphere is about 11% and 14% respectively. The calculation indicates that the curvature difference of the profile of the square lens in the orthogonal and diagonal direction is 5.5%. The roughness of the lens is measured as approximately 6 nm.