The interactions between pairs of cells and within multicellular assemblies are critical to many biological processes such as intercellular communication, tissue and organ formation, immunological reactions, and cancer metastasis. The ability to precisely control the position of cells relative to one another and within larger cellular assemblies will enable the investigation and characterization of phenomena not currently accessible by conventional in vitro methods. We present a versatile surface acoustic wave technique that is capable of controlling the intercellular distance and spatial arrangement of cells with micrometer level resolution. This technique is, to our knowledge, among the first of its kind to marry high precision and high throughput into a single extremely versatile and wholly biocompatible technology. We demonstrated the capabilities of the system to precisely control intercellular distance, assemble cells with defined geometries, maintain cellular assemblies in suspension, and translate these suspended assemblies to adherent states, all in a contactless, biocompatible manner. As an example of the power of this system, this technology was used to quantitatively investigate the gap junctional intercellular communication in several homotypic and heterotypic populations by visualizing the transfer of fluorescent dye between cells.cell-cell interaction | intercellular communication | surface acoustic waves | acoustic tweezers | acoustofluidics M ulticellular systems rely on the interaction between cells to coordinate cell signaling and regulate cell functions. Understanding the mechanism and process of cell-cell interaction is critical to many physiological and pathological processes, such as embryogenesis, differentiation, cancer metastasis, immunological interactions, and diabetes (1-3). Despite significant advances in this field, to further understand how cells interact and communicate with each other, a robust, biocompatible method to precisely control the spatial and temporal association of cells and to create defined cellular assemblies is urgently needed (4). Although several methods have been used to pattern cells, limitations still exist for the demonstrated methods including those that make use of optical, electrical, magnetic, hydrodynamic, and contact printing technologies (5-9). Firstly, most of the methods require modification of the cell's native state. The magnetic assembly method, for example, requires cells to be labeled with magnetic probes. Dielectrophoresis typically requires the use of a special medium (e.g., nonconductive) which may lack essential nutrients or have biophysical properties (such as the osmolality) that may adversely affect cell growth or physiology (6). Optical tweezers provide a label-free and contactless approach, but typically require high laser power to manipulate cells, leading to a high risk of cell damage (5). Secondly, the working principles of the existing technologies mostly preclude the combination of high precision and high throughput into a single ...

Herpes simplex virus thymidine kinase (HSVtk)/ganciclovir (GCV) viral-directed enzyme prodrug gene therapy causes potent, tumor-selective cytotoxicity in animal models in which HSV-tk gene transduction is limited to a minority of tumor cells. The passage of toxic molecules from HSV-tk+ cells to neighboring HSV-tk-cells during GCV therapy is one mechanism that may account for this "bystander" cytotoxicity. To investigate whether gap junction-mediated intercellular coupling could mediate this bystander effect, we used a flow cytometry assay to quantitate the extent of heterocellular coupling between HSV-tk+ murine fibroblasts and both rodent and human tumor cell lines. Bystander tumor cytotoxicity during GCV treatment in a coculture assay was highly correlated (P < 0.001) with the extent of gap junctionmediated coupling. These (HSV-tk) to tumor cells (3-7). Transduced cells become sensitive to ganciclovir (GCV), a guanine analog that causes premature chain termination when incorporated into replicating DNA, thereby disrupting cellular proliferation (8, 9). The death of tumor cells not transduced with the HSV-tk transgene is called the "bystander effect" (3).Bystander cell killing occurs in cocultures of HSV-tk+ and unmodified HSV-tk-tumor cells when as few as 3% of cells express HSV-tk (10). Bystander killing in the HSV-tk/GCV system requires direct contact between HSV-tk+ cells and bystander cells (11) and is not mediated by soluble factors (10, 12). Toxic GCV metabolites transferred through gap junctions linking neighboring cells has been proposed as one possible mechanism of bystander killing (11,13 11071The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.

The Pelorus Mark III Stereotactic System is a commercially available device for CT-guided stereotactic brain biopsy in people. With relatively minor modifications, this device was used to safely and accurately perform CT-guided stereotactic brain biopsies in 50 dogs with intracranial lesions. Modifications were necessary to accommodate a 90 degree shift in orientation of the canine head compared to the human head during the imaging phase of the procedure, and to facilitate other phases of the biopsy procedure that are affected by the uneven and variable topography of the canine skull. Description of a typical CT-guided brain biopsy procedure in dogs using the modified Pelorus Mark III Stereotactic System is provided. Accuracy of biopsy needle placement was determined by comparing the x, y and z coordinates of the biopsy target site with the actual coordinates of the needle tip on CT images. Mean needle placement error was 3.5 +/- 1.6 mm. Needle placement error was not significantly related to operator experience, dog size (body weight), or needle path length, however, needle placement error was significantly affected by lesion location.