Lee E. Weiss

Dynamic sensor-based control of robots with visual feedback

Sensor-based robot control may be viewed as a hierarchical structure with multiple observers. Actuator, feature-based, and recognition observers provide the basis for multilevel feedback control at the actuator, sensor, and world coordinate frame levels, respectively. The analysis and design of feature-based control strategies to achieve consistent dynamic performance is addressed. For vision sensors, such an image-based visual servo control is shown to provide stable and consistent dynamic control within local regimes of the recognition observer. Simulation studies of two- and three-degree-of-freedom systems show the application of an adaptive control algorithm to overcome unknown and nonlinear relations in the feature to world space mapping.

In vitro analysis of biodegradable polymer blend/hydroxyapatite composites for bone tissue engineering

Blends of biodegradable polymers, poly(caprolactone) and poly(D, L-lactic-co-glycolic acid), have been examined as scaffolds for applications in bone tissue engineering. Hydroxyapatite granules have been incorporated into the blends and porous discs were prepared. Mechanical properties and degradation rates in vitro of the composites were determined. The discs were seeded with rabbit bone marrow or cultured bone marrow stromal cells and incubated under physiological conditions. Polymer/ceramic scaffolds supported cell growth throughout the scaffold for 8 weeks. Scanning and transmission electron microscopy, and histological analyses were used to characterize the seeded composites. This study suggests the feasibility of using novel polymer/ceramic composites as scaffold in bone tissue engineering applications.

Cell Population Tracking and Lineage Construction with Spatiotemporal Context

Automated visual-tracking of cell populations in vitro using time-lapse phase contrast microscopy enables quantitative, systematic, and high-throughput measurements of cell behaviors. These measurements include the spatiotemporal quantification of cell migration, mitosis, apoptosis, and the reconstruction of cell lineages. The combination of low signal-to-noise ratio of phase contrast microscopy images, high and varying densities of the cell cultures, topological complexities of cell shapes, and wide range of cell behaviors poses many challenges to existing tracking techniques. This paper presents a fully automated multi-target tracking system that can efficiently cope with these challenges while simultaneously tracking and analyzing thousands of cells observed using time-lapse phase contrast microscopy. The system combines bottom-up and top-down image analysis by integrating multiple collaborative modules, which exploit a fast geometric active contour tracker in conjunction with adaptive interacting multiple models (IMM) motion filtering and spatiotemporal trajectory optimization. The system, which was tested using a variety of cell populations, achieved tracking accuracy in the range of 86.9-92.5%.

Microenvironments engineered by inkjet bioprinting spatially direct adult stem cells toward muscle- and bone-like subpopulations

In vivo, growth factors exist both as soluble and as solid‐phase molecules, immobilized to cell surfaces and within the extracellular matrix. We used this rationale to develop more biologically relevant approaches to study stem cell behaviors. We engineered stem cell microenvironments using inkjet bioprinting technology to create spatially defined patterns of immobilized growth factors. Using this approach, we engineered cell fate toward the osteogenic lineage in register to printed patterns of bone morphogenetic protein (BMP) 2 contained within a population of primary muscle‐derived stem cells (MDSCs) isolated from adult mice. This patterning approach was conducive to patterning the MDSCs into subpopulations of osteogenic or myogenic cells simultaneously on the same chip. When cells were cultured under myogenic conditions on BMP‐2 patterns, cells on pattern differentiated toward the osteogenic lineage, whereas cells off pattern differentiated toward the myogenic lineage. Time‐lapse microscopy was used to visualize the formation of multinucleated myotubes, and immunocytochemistry was used to demonstrate expression of myosin heavy chain (fast) in cells off BMP‐2 pattern. This work provides proof‐of‐concept for engineering spatially controlled multilineage differentiation of stem cells using patterns of immobilized growth factors. This approach may be useful for understanding cell behaviors to immobilized biological patterns and could have potential applications for regenerative medicine.

Structured highlight inspection of specular surfaces

An approach to illumination and imaging of specular surfaces that yields three-dimensional shape information is described. The structured highlight approach uses a scanned array of point sources and images of the resulting reflected highlights to compute local surface height and orientation. A prototype structured highlight inspection system, called SHINY, has been implemented. SHINY demonstrates the determination of surface shape for several test objects including solder joints. The current SHINY system makes the distant-source assumption and requires only one camera. A stereo structured highlight system using two cameras is proposed to determine surface-element orientation for objects in a much larger field of view. Analysis and description of the algorithms are included. The proposed structured highlight techniques are promising for many industrial tasks. >

Tissue engineering with the aid of inkjet printers

Tissue engineering holds the promise to create revolutionary new therapies for tissue and organ regeneration. This emerging field is extremely broad and eclectic in its various approaches. However, all strategies being developed are based on the therapeutic delivery of one or more of the following types of tissue building-blocks: cells; extracellular matrices or scaffolds; and hormones or other signaling molecules. So far, most work has used essentially homogenous combinations of these components, with subsequent self-organization to impart some level of tissue functionality occurring during in vitro culture or after transplantation. Emerging ‘bioprinting’ methodologies are being investigated to create tissue engineered constructs initially with more defined spatial organization, motivated by the hypothesis that biomimetic patterns can achieve improved therapeutic outcomes. Bioprinting based on inkjet and related printing technologies can be used to fabricate persistent biomimetic patterns that can be used both to study the underlying biology of tissue regeneration and potentially be translated into effective clinical therapies. However, recapitulating nature at even the most primitive levels such that printed cells, extracellular matrices and hormones become integrated into hierarchical, spatially organized three-dimensional tissue structures with appropriate functionality remains a significant challenge.

Shape deposition manufacturing of heterogeneous structures

Abstract To date, the most widely recognized advantage of the layered manufacturing methodology is the relative ease of automatically planning and executing the fabrication of complex geometric shapes. But building shapes using selective material-additive processes has a second, far-reaching advantage—the creation of heterogeneous structures composed of multimaterial regions and with prefabricated devices embedded into the structure. The capability to fabricate heterogeneous structures is important because it enables the realization of new, complex designs. Shape deposition manufacturing, a layered manufacturing process described in this paper, addresses how to build multimaterial, embedded structures such as advanced tooling and embedded electronic devices.

Specular surface inspection using structured highlight and Gaussian images

The structured highlight inspection method uses an array of point sources to illuminate a specular object surface. The point sources are scanned, and highlights on the object surface resulting from each source are used to derive local surface orientation information. The extended Gaussian image (EGI) is obtained by placing at each point on a Gaussian sphere a mass proportional to the area of elements on the object surface that have a specific orientation. The EGI summarizes shape properties of the object surface and can be efficiently calculated from structured highlight data without surface reconstruction. Features of the estimated EGI including areas, moments, principal axes, homogeneity measures, and polygonality can be used as the basis for classification and inspection. The structured highlight inspection system (SHINY) has been implemented using a hemisphere of 127 point sources. The SHINY system uses a binary coding scheme to make the scanning of point sources efficient. Experiments have used the SHINY system and EGI features for the inspection and classification of surface-mounted-solder joints. >

Bioprinting of growth factors onto aligned sub-micron fibrous scaffolds for simultaneous control of cell differentiation and alignment

The capability to spatially control stem cell orientation and differentiation simultaneously using a combination of geometric cues that mimic structural aspects of native extracellular matrix (ECM) and biochemical cues such as ECM-bound growth factors (GFs) is important for understanding the organization and function of musculoskeletal tissues. Herein, oriented sub-micron fibers, which are morphologically similar to musculoskeletal ECM, were spatially patterned with GFs using an inkjet-based bioprinter to create geometric and biochemical cues that direct musculoskeletal cell alignment and differentiation in vitro in registration with fiber orientation and printed patterns, respectively. Sub-micron polystyrene fibers (diameter ~ 655 nm) were fabricated using a Spinneret-based Tunable Engineered Parameters (STEP) technique and coated with serum or fibrin. The fibers were subsequently patterned with tendon-promoting fibroblast growth factor-2 (FGF-2) or bone-promoting bone morphogenetic protein-2 (BMP-2) prior to seeding with mouse C2C12 myoblasts or C3H10T1/2 mesenchymal fibroblasts. Unprinted regions of STEP fibers showed myocyte differentiation while printed FGF-2 and BMP-2 patterns promoted tenocyte and osteoblast fates, respectively, and inhibited myocyte differentiation. Additionally, cells aligned along the fiber length. Functionalizing oriented sub-micron fibers with printed GFs provides instructive cues to spatially control cell fate and alignment to mimic native tissue organization and may have applications in regenerative medicine.

Characterization of osteoblast-like behavior of cultured bone marrow stromal cells on various polymer surfaces

The creation of novel bone substitutes requires a detailed understanding of the interaction between cells and materials. This study was designed to test certain polymers, specifically poly(caprolactone) (PCL), poly(D,L-lactic-CO-glycolic acid) (PLGA), and combinations of these polymers for their ability to support bone marrow stromal cell proliferation and differentiation. Bone marrow stromal cells were cultured from New Zealand White rabbits and were seeded onto glass slides coated with a thin layer of PCL, PLGA, and combinations of these two polymers in both a 40:60 and a 10:90 ratio. Growth curves were compared. At the end of 2 weeks, the cells were stained for both matrix mineralization and alkaline phosphatase activity. There was no statistically significant difference in growth rate of the cells on any polymer or polymer combination. However, there was a striking difference in Von Kossa staining and alkaline phosphatase staining. Cells on PCL did not show Von Kossa staining or alkaline phosphatase staining. However, in the 40:60 and 10:90 blends, there was both positive Von Kossa and alkaline phosphatase staining. These data indicate that PCL alone may not be a satisfactory material for the creation of a bone substitute. However, it may be used in combination with PLGA for the creation of a bone substitute material.