text:
|Publication number
|Publication type
|Application number
|Publication date
|Dec 5, 2006
|Filing date
|Mar 8, 2002
|Priority date
|Mar 8, 2001
|Fee status
|Also published as
|Publication number
|Jose de la Torre-Bueno, Peter Salamon
|Original Assignee
|Clarient, Inc.
|Export Citation
|BiBTeX, EndNote, RefMan
|Patent Citations (11), Non-Patent Citations (4), Referenced by (10), Classifications (25), Legal Events (9)
|External Links: USPTO, USPTO Assignment, Espacenet
This application claims priority to Provisional Application Ser. No. 60/274,688, filed Mar. 8, 2001.
1. Field of the Invention
The subject matter described herein relates generally to labeling samples in an array.
2. Background of the Invention
An array is an ordered arrangement of any kind of subject matter. In the biological sciences, an array is generally a two-dimensional arrangement of samples placed upon a support structure. Samples can include, but are not limited to, nucleic acids, proteins, molecules, cells, tissues and any combination thereof. These arrays allow for the efficient and rapid processing of large numbers of samples, allowing laboratories to process thousands of samples a day. For example, a microplate configured with a four by four matrix of biosites or samples in each of the 96 wells of a microtiter plate would be able to perform a total of 1536 nearly simultaneous tests utilizing a proximal CCD imager. A microplate configured with a 15 by 15 matrix of samples in each of the 96 wells enables a total of 21,600 nearly simultaneous reactions to be processed.
Arrays of samples are used in many forms and for many areas of science, including, but not limited to, 96 well plates or slides for combinatorial chemistry, multi-well carriers for synthesizers, and papers, such as nitrocellulose and nylon, for hybridization reactions. Array technology can be used with almost any clinical or research protocol. For example, in screening libraries, the library, consisting of recombinant clones or molecules, can be placed in two-dimensional arrays on supports, examples of supports can include a microtiter plate or microscope slide. Each clone or molecule can be identified by the identity of the plate and the clone or molecule location (row and column) on that plate. The arrayed libraries can then be used for many applications such as screening for a specific gene of interest or for identifying potential lead compounds for treating diseases. Arrays can also be used to diagnosis diseases as well as synthesize nucleic acids, polypeptides, and chemical compounds. As another example, arrays of tissue can be arranged on a microscope slide to simultaneously test an experimental treatment for diseased tissues.
Current designs rely on the wells or dots being in predictable positions so they can be processed or read by robotic equipment. If, for any reason, the positioning is not as expected, a possible response by a computerized system is to shut down processing of the array. Augmenting a vision system with a method for accommodating irregular arrays would allow robotic systems of this type to recover from minor positioning errors.
Examples of arrays are shown in
Once the slides of tissue are ready to be analyzed, the slides can be scanned to produce digitized images. The digitized images of the stained slides can then be automatically processed. However, before processing proceeds, it may be desirable to identify the regions of the image corresponding to each tissue sample. The digitized image may be analyzed to locate connected regions forming the tissue samples and their centroids. Once the tissue samples and/or their centroids are located, they can be assigned coordinates. Assigning coordinates to the samples may facilitate later data analysis and allow one to return to a tissue sample of interest. While the identification of the samples and/or their centroids can be preformed prior to processing the samples in a desired protocol, identification of the samples and their centroids can also be done after the processing of the samples.
In identifying and assigning coordinates to the samples, several image irregularities may hinder machine determination of the row and column coordinates of the centroids. Reasons for the irregularities range from human error to shearing of the paraffin during handling. The irregularities make it difficult to know with certainty the correct labeling in the areas of these irregularities.
Even when a guide is employed, the regularity with which the tissue can be placed into the paraffin may be less than perfect. The tissue sections may be subject to deformation during the transfer to the slide, and some samples may fail to adhere to the slide. In cases where samples in liquid are applied to a support of the array, the liquid dispenser may inadvertently omit samples, for example, failure to pick up the liquid sample, or introduce extraneous "samples," for example, the liquid sample may inadvertently drip from the liquid dispenser or stray marks are introduced by mishandling the slide. Moreover, rows and/or columns, or portions thereof, of samples may be intentionally omitted to separate different groups of samples.
The digitization process introduces further noise to challenge the row and column identification. The noise level may be sufficiently high that a simple round to the nearest integer in
ncols*X/Xmax (1)
does not reveal the centroid's column coordinate, where X is the x coordinate of the centroid in question, Xmax is the greatest x coordinate of all the centroids, and ncols is the number of columns on the slide.
The problem of sparse data is evident in various regions of
In accordance with the present invention, a method for assigning coordinates to a centroid in an array would include the steps of providing an array to be analyzed; selecting a starting centroid within the array by using a local lattice structure and determining a quality of fitness for each centroid; assigning coordinates to the starting centroid; and diffusing outward to determine and assign coordinates to neighboring centroids using the lattice structure. While the present invention can be used with arrays whose samples are in perfect alignment, the present invention is especially useful when the array is irregular. The present invention can analyze all the centroids of the array or the centroids located on portions of the array.
In some embodiments of the invention, a quality of fitness value is calculated for a given centroid, (xc, yc). The quality of fitness value being determined by the equation
A local lattice structure can be a grid having a center centroid (xc, yc). N is the number of neighboring centroids (xk, yk) and va and vb are adjusted to minimize the sum of the squared distances from an actual position of each of the neighboring centroids to the nearest lattice point. The lattice points relative to the center centroid can be calculated by the equation
(x grid , y grid)=(x c , y c)+K�v a +L�v b
wherein K and L are integers and va and vb are two lattice vectors that describe the lattice structure. Each neighboring centroid can be associated with a point of the lattice by minimizing
(x k −x grid)2+(y k −y grid)2
over K and L for any given value of va and vb for the grid as defined by
(x grid , y grid)=(x c , y c)+K�v a +L�v b.
The grid of the lattice structure can be of any size, limited only in that the grid must accommodate a center centroid and neighboring centroids. Thus, the grid can be a three-by-three grid having eight neighboring lattice points, a five-by-five grid having 24 neighboring lattice points, a seven-by-seven grid having 48 neighboring lattice points, and so on.
In some embodiments, the method includes a step of identifying (labeling) suspicious points (centroids) by determining whether the quality of fitness of a given centroid is greater than a first tolerance level or whether the lattice vectors associated with the centroid relative to the lattice vectors associated with the starting centroid is greater than a second tolerance level.
The step of identifying the suspicious point can be executed before the step of determining the starting centroid or before the step of diffusing outward. Additionally the step of identifying the suspicious point can be done periodically throughout the diffusing step, or after each centroid has been selected as the center centroid and the centroids on the grid have been assigned coordinates.
The first tolerance level, for identifying suspicious points, is ε1 and is defined by
ε1 =N(Mδ 1)2
wherein N is a number of neighbors, M is a mean lattice spacing of the best lattice structure as defined by
and δ1 represents the average deviation between a lattice point and a neighboring centroid in units of M.
The second tolerance level is defined by
wherein ε2=δ2 and δ2 is a user specified tolerance threshold.
In some embodiments, the step of diffusing outward includes selecting one of the neighboring centroid (xk, yk) as a new center centroid. To be a new center centroid, the centroid must not have been identified (labeled) as suspicious or ambiguous, or previously been a center centroid. The method also includes the steps of assigning the new center centroid a relative coordinate (Ik, Jk); providing a lattice structure comprising a grid; locating a neighboring centroid to the new center centroid closest to a point on the grid of the lattice structure using an equation
(x grid , y grid)=(x k , y k)+Kv a k +Lv b k;
and assigning the neighboring centroid a relevant coordinate using the equation
(I new ,J new)=(I k ,J k)+(K,L).
The neighboring centroid, however, may not be assigned relative coordinates if the distance as defined by √{square root over ((x−xgrid)2+(y−ygrid)2)}{square root over ((x−xgrid)2+(y−ygrid)2)} is greater than a third tolerance level defined as ε3. The third user specified tolerance level δ3 is defined by
In some embodiments, the grid of the lattice structure used in the diffusing step is of a different size than the grid used for determining a starting centroid. In other embodiments, they are the same.
In some embodiments, where the neighboring centroid has already been assigned coordinates, if the previously assigned coordinates do not match the new coordinates, then the centroid will be identified as ambiguous.
In some embodiments, the diffusion steps are repeated until all the centroids in the array, not labeled suspicious or ambiguous, have been the center centroid.
In some embodiments the relative coordinates of the centroids in the array are translated to absolute coordinates. The absolute coordinates can be calculated from the equation
(O, P)=(I, J)−(I min , J min)+(1, 1)
wherein Imin and Jmin represent the smallest assigned I and J values.
In some embodiments, the method further includes the step of outputting data. The data includes the centroids being labeled either with coordinates, identified as suspicious or ambiguous, or remain unlabeled.
In some embodiments, the method for assigning coordinates to centroids in an array is used in an automated system. The method would include the steps of scanning the array; and inputting the scanned image to a receiver for processing the array according to the methods of the invention.
In some embodiments, an array of biological samples is placed in an array carrier. The array carriers are loaded into an input hopper of an automated system. The operator may then enter data identifying the size, shape and location of a scan area on each array, or, preferably, the system automatically locates a scan area for each array during array processing. An operator can then activate the system for slide processing. Alternatively, the processing parameters of the array may be identified by a bar code present on the array or array carrier. Arrays can be made from slides, microtitre plates, multi-well trays, and paper, for example, nitrocellulose or nylon. Other material used in the production of arrays are well known in the art. For example, methods of preparing an array can be found in U.S. Pat. No. 6,312,960, U.S. Pat. No. 6,203,758, U.S. Pat. No. 6,103,479, and U.S. Pat. No. 5,770,151, all of which are incorporated by reference in their entirety.
At system activation, an array carrier can be positioned on an X–Y stage, the entire array is then rapidly scanned, thereby acquiring an image for processing and detecting objects of interest, samples and their centroids. Color, size, density, shape, and texture of the samples can be used to identify these objects of interest. The location of each candidate object of interest can then be stored for further processing, such as assigning coordinates to the objects of interest.
Embodiments of an apparatus of the invention can include a computer program, stored on a computer-readable medium, for assigning coordinates to a centroid on an array, the computer program comprising instructions for causing a computer system to provide an image of an array; select a starting centroid within the array by using a lattice structure and determine a quality of fitness for each centroid; assign coordinates to the starting centroid; and diffuse outward to determine and assign coordinates to a neighboring centroid using the lattice structure. It is contemplated that the method of assigning coordinates can be performed by an algorithm.
Also contemplated is an apparatus, for assigning coordinates to samples (objects of interest) in an array, including a computer having at least one system processor with image processing capability, a computer monitor, an input device, a power supply, and an optical sensing device for acquiring images. The apparatus can also include input and output mechanisms for multiple array analysis and storage.
As used herein and in the appended claims, the singular forms "a," "an," and "the" include plural references unless the context clearly dictates otherwise. Thus, for example, reference to "an array" can include a plurality of arrays and a reference to "the centroid" can include reference to one or more centroids, and so forth.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood to one of ordinary skill in the art to which this invention belongs. Although any method, device and material similar or equivalent to those described herein can be used in the practice of the invention, the preferred methods, devices and materials are now described.
The apparatus and method described herein, for assigning coordinates to samples of an array, work from an initial list of centroids. A centroid represents the center of mass of a sample or a putative sample; therefore, in the analysis, a centroid corresponds to a sample of the array. In practicing the invention, the apparatus and method associate to each centroid a set of coordinates, for example, a row and a column in the array, or identify the centroid as a suspicious or ambiguous point that should be excluded from the regular array.
The inventive apparatus and method for labeling rows and columns in arrays of samples can accommodate the irregularities of the array described above. The apparatus and method proceed via a sequence of deductions concerning the lattice structure of the array. The apparatus and method begin from a region that conforms well to a local lattice structure and spreads out from this initial seed.
The method selects a best local lattice structure for the centroids and calculates a measure of the quality of fitness of the lattice to the centroids 130. The calculated values are then sorted by their quality of fitness to determine the centroid having the best quality of fitness value and defining that centroid as the starting point 140.
The measure of the quality of fitness of a lattice structure for a given centroid is computed as follows. The lattice points relative to a center centroid (xc, yc) are given by the equation
(x grid , y grid)=(x c , y c)+K�v a +L�v b (2)
where K and L are integers and va and vb are the two lattice vectors that describe the local lattice and that remain to be determined as part of the fit. In the case of a local lattice having an three-by three grid, the eight closest centroids N8={(xk, yk), k=1, . . . 8}, are each associated with a node, or point, on the model grid by minimizing the equation
(x k −x grid)2+(y k −y grid)2 (3)
over the values of K and L for any given values of va and vb, where (xgrid, ygrid) are as defined in equation (2). The grid parameters va and vb are then adjusted to minimize the sum of the squared distances from the actual positions of the eight centroids closest to (xc, yc) and their nearest lattice points. Specifically, the quality of fitness, Fit(xc, yc), for a given centroid, (xc, yc) is determined by the equation
where N is the number of neighboring centroids, in this case eight. The sum of these eight squared distances is the measure of the quality of fitness of a region's local lattice match.
Given reasonably close starting values of va and vb, a local optimization can be carried out analytically. The choice of K and L for a given center centroid (xc, yc), its neighboring centroids (xk, yk), and lattice vectors va and vb is achieved by expressing the displacement (xk−xc, yk−yc) as a linear combination of va and vb, and rounding the coefficients to the nearest integers. Using these integers Kk and Lk for the (xgrid(k), ygrid(k)) positions, the resulting objective function is quadratic in the four unknowns va=(vax, vay) and vb, =(vbx, vby). Setting the four partial derivatives of the objective function in equation (4) equal to zero, gives four linear equations which may be solved for the optimal values of va and vb. The solution determines the horizontal and vertical vectors that generate the best model grid for the region. The starting values of va and vb are
v a=(X max /n cols, 0) and v b=(0, Y max /n rows).
Once a region with the best quality of fitness value is found, its va and vb are used as initial values for the next optimization.
The previous paragraphs describe how each centroid is assigned an associated pair of lattice vectors va and vb and a quality of fitness value. The starting point of the method is the centroid with the best quality of fitness value. This starting centroid is assigned relative row and column coordinates (I,J)=(0,0) and its lattice vectors are denoted by va* and vb*.
Referring to the embodiment shown in
Suspicious points can be identified by either (a) having a quality of fitness value as determined by equation 4 above some tolerance level ε1, or (b) having a value when the local lattice vector associated with the centroid in question is compared to the local lattice vectors associated with the starting centroid (which has been determined to have the best quality of fitness value) above some tolerance level ε2. The tolerance levels can be user specified.
In one embodiment, the tolerance levels can specify limiting values of certain quantities calculated from the (x,y) coordinates of the centroids and, as such, may be highly dependent on the scale used in digitizing the image. To eliminate this dependence, the values of these tolerance levels, ε1 and ε2, are calculated from user-specified tolerances δi, i=1 and 2 which are measured in units of a lattice spacing.
To rescale from units of a lattice spacing to the units used in the digitized grid, it is convenient to define a parameter M, which represents the mean lattice spacing, as
where va* and vb* are the lattice vectors of the starting centroid, having been determined that the centroid had the best quality of fitness value. It is also possible to define M by using the scales of Δx and Δy.
Tolerance level ε1 can be defined as
ε1=N(Mδ 1)2
where N is the number of neighboring centroids, which would be eight for a three-by-three grid. For a centroid to be identified as suspicious, its quality of fitness is greater than the tolerance level ε1
ε1<Fit(x c , y c)
and Fit(xc, yc) is a sum of N squared deviations and δ1 represents the average deviation between a lattice point and a neighboring centroid in units of M. The user can also specify that if the quality of fitness is equal to ε1, the centroid should also be labeled as suspicious.
The second tolerance level, ε2, is used to identify suspicious points on the grounds that their lattice vectors deviate too much from the ideal. In the method, this can be measured by
Since ε2, specifies a fractional difference, then
This makes δ2 a fractional threshold comparing the size of the locally optimized lattice of the centroid to the best lattice of the starting centroid. This test condition serves to eliminate harmonics. If a set of points is well described by the lattice vectors va, vb, then they are also well described by va/2 and vb/2. These locally optimal solutions are called harmonics. If the optimized lattice is too small, then the lattice vectors correspond to higher harmonics. Similarly, if the lattice is too large it is called lower harmonics.
Once all the centroids have been evaluated in order to identify suspicious centroids, the diffusion step proceeds 160. Alternatively or additionally, the method may provide for re-evaluating the centroids periodically to note any changes in the tolerance values. In other words, as the diffusion step proceeds, a centroid may be a neighboring centroid in several lattice structures. As more information is gathered, the tolerance measurements for a centroid may be re-calculated to see if additional information has changed the tolerance measures below the applicable tolerance threshold levels, ε1 and ε2, to justify removal of the suspicious label. This "re-evaluation" of a suspicious point can be done after the assignment of a coordinate, or before the selection of a new center centroid. It can also be done periodically during the diffusion process.
In the diffusion process 160, the method provides for selecting a centroid (xk, yk) to be used as the next center of a new local lattice and then assigning row and column coordinates to all the points lying sufficiently near the lattice point, such as those found in a five-by-five grid having a center at (xk, yk) 170. The new center must fulfill four criteria:
Among the centroids that fulfill these four conditions, the one which lies closest to the starting center (I,J)=(0,0) can be used as the next center centroid.
(x grid , y grid)=(x k , y k)+Kv a k +Lv b k
and tentatively assigned the coordinates
(I new ,J new)=(I k ,J k)+(K,L).
These tentatively assigned coordinates are actually assigned to this centroid only if no previous coordinates have been assigned to this point and the distance
√{square root over ((x−x grid)2+(y−y grid)2)}{square root over ((x−x grid)2+(y−y grid)2)}
is less than a third tolerance level ε3. If coordinates have previously been assigned and they do not match the newly calculated coordinates, then the centroid is identified as "ambiguous" and, thus, will not be used as a center.
A user specified tolerance level ε3 is used in the equation
ε3>√{square root over ((x−x grid)2+(y−y grid)2)}{square root over ((x−x grid)2+(y−y grid)2)}
to decide whether the neighboring centroid (x,y) of a center centroid should be labeled based on its proximity to the nearest grid point emanating from the center centroid. Therefore, if the third tolerance value of the neighboring centroid is less than ε3 and no previous coordinates have been assigned it, it will be assigned a coordinate. Defining δ3 by
makes δ3 scale independent.
The diffusion process continues until all eligible centroids have been used as the center centroid 180. The relative row and column coordinates (I,J) can then be translated to absolute row and column coordinates by the following equation
(O, P)=(I, J)−(I min , J min)+(1, 1)
where Imin and Jmin represent the smallest assigned I and J values 190. Besides the row and column assignments, the output of the present method can include lists of suspicious, ambiguous, and unlabeled centroids 200.
The various techniques, methods, and aspects of the invention described above can be implemented in part or in whole using computer-based systems and methods. Additionally, computer-based systems and methods can be used to augment or enhance the functionality described above, increase the speed at which the functions can be performed, and provide additional features and aspects as a part of or in addition to those of the invention described elsewhere in this document, various computer-based systems, methods and implementations in accordance with the above-described technology are presented below.
An apparatus of the invention may include the use of the methods of the invention in conjunction with any of the computer-based systems described herein. The computer-based system may include the means for inputting data either manually or automatically. The system can include devices for scanning the array, or otherwise inputting data about the array, digitizing the image, storing data, processing the data according to the methods described herein, outputting data, or any one of these functions. Any or all of these functions may be automated.
The processor-based system can include a main memory, preferably random access memory (RAM), and can also include a secondary memory. The secondary memory can include, for example, a hard disk drive and/or a removable storage drive, representing a floppy disk drive, a magnetic tape drive, an optical disk drive, etc. The removable storage drive reads from and/or writes to a removable storage medium. Removable storage media represents a floppy disk magnetic tape, optical disk, etc., which is read by and written to by removable storage drive. As will be appreciated, the removable storage media includes a computer usable storage medium having stored therein computer software and/or data.
In alternative embodiments, secondary memory may include other similar means for allowing computer programs or other instructions to be loaded into a computer system. Such means can include, for example, a removable storage unit and an interface. Examples of such can include a program cartridge and cartridge interface (such as the found in video game devices), a movable memory chip (such as an EPROM, or PROM) and associated socket, and other removable storage units and interfaces which allow software and data to be transferred from the removable storage unit to the computer system.
The computer system can also include a communications interface. Communications interfaces allow software and data to be transferred between computer system and external devices. Examples of communications interfaces can include a modem, a network interface (such as, for example, an Ethernet card), a communications port, a PCMCIA slot and card, etc. Software and data transferred via a communications interface are in the form of signals which can be electronic, electromagnetic, optical or other signals capable of being received by a communications interface. These signals are provided to communications interface via a channel capable of carrying signals and can be implemented using a wireless medium, wire or cable, fiber optics or other communications medium. Some examples of a channel can include a phone line, a cellular phone link, an RF link, a network interface, and other communications channels.
In this document, the terms "computer program medium" and "computer usable medium" are used to generally refer to media such as a removable storage device, a disk capable of installation in a disk drive, and signals on a channel. These computer program products are means for providing software or program instructions to a computer system.
Computer programs (also called computer control logic) are stored in main memory and/or secondary memory. Computer programs can also be received via a communications interface. Such computer programs, when executed, enable the computer system to perform the features of the present invention as discussed herein. In particular, the computer programs, when executed, enable the processor to perform the features of the present invention. Accordingly, such computer programs represent controllers of the computer system.
In an embodiment where the elements are implemented using software, the software may be stored in, or transmitted via, a computer program product and loaded into a computer system using a removable storage drive, hard drive or communications interface. The control logic (software), when executed by the processor, causes the processor to perform the functions of the invention as described herein.
In another embodiment, the elements are implemented primarily in hardware using, for example, hardware components such as PALs, application specific integrated circuits (ASICs) or other hardware components. Implementation of a hardware state machine so as to perform the functions described herein will be apparent to person skilled in the relevant art(s). In yet another embodiment, elements are implanted using a combination of both hardware and software.
In another embodiment, the computer-based methods can be accessed or implemented over the World Wide Web by providing access via a Web Page to the methods of the present invention.
In one embodiment, the X–Y stage is moved to scan the image of the array. Alternatively or additionally, it is the microscope subsystem 32 and/or optical sensing device 42 that is moved to scan the array. The apparatus may also include a Z stage or focus stage 46 under control of the system processor 23 to provide displacement of the microscope subsystem in the Z plane for focusing. The microscope subsystem 32 can further include a motorized objective turret 44 for selection of objectives to view the array. In other embodiments, the array is scanned in a manner similar to that of electronic scanners, where an "eye" picks up images and digitalizes it as the "eye" is moved along, directly or indirectly, to the surface of the array.
One purpose of the apparatus 10 is for the unattended automatic scanning of prepared arrays for the detection of samples in the array. The apparatus 10 can then automatically locate and assign coordinates to the samples in the array. A number of stains can be used, if desired, to distinguish the samples from the background. Thus, in one aspect, the samples can be prepared with a reagent to obtain a colored insoluble precipitate. The apparatus of the present invention being used to detect this precipitate as a sample. Other means of distinguishing samples from background include, but are not limited to texture and density analysis. Methods and apparatus of automated image analysis and detection of objects of interest, e.g., samples on slides, are provided in U.S. patent application Ser. No. 09/344, 308, filed Jun. 24, 1999; U.S. patent application Ser. No. 09/495,461, filed Feb. 1, 2000; and U.S. patent application Ser. No. 09/616,817, filed Jul. 12, 2002, all of which are explicitly incorporated by reference in their entirety.
During operation of the apparatus 10, a user can mount the prepared arrays onto the input hopper 16. The hopper 16 may utilize an automatic feed mechanism 37. The user can then specify the size, shape and location of the area to be scanned or, alternatively, the system can automatically scan the array. The operator can command the apparatus to begin automated scanning of the array through a graphical user interface. A bar code label can be affixed to the array or array carrier and read by a bar code reader 33 during this loading operation.
The apparatus stores an image of the array for later analysis or review by a user. All results and images can be stored to a storage device 21 such as a removable hard drive, DAT tape, local hard drive, optical disk, or transmitted to a remote site for review or storage. The stored images for each slide can be viewed in a mosaic of images for further review.
The following examples are provided to illustrate the practice of the instant invention and in no way limit the scope of the invention.
In evaluating the experimental values for δs, Table 1 below shows the range of parameter values that can produce satisfactory performance using a method of the invention. The table also summarizes the meaning and use of each of the parameters.
Mean deviation of
Decide whether
neighboring points
to use centroid
from the local lattice
as center
Fractional deviation
Decide whether
of local lattice
to use centroid
vectors from best
as center
lattice vectors
Deviation of one
Decide whether
neighbor to nearby
to assign label
grid point
to neighbor
The method described above performed satisfactorily on all datasets considered. The arrays of
In evaluating the method of the invention, it is useful to introduce three complementary ways of visualizing the results obtained from practicing the method of the invention. The first is the finished version of the lattices generated during the diffusion phase.
The lattice visualization shows each edge (the line segment connecting two adjacent points in a row or column) as represented in six different local lattices. These lattices are well aligned if the six line segments appear as a single line segment. Small misalignments cause the line segment to appear thicker, and large misalignments produce shadows or completely separate traces. As a result, this way of displaying the data can explain any difficulties encountered by the algorithm. The "ambiguous" labels are due to the very slanted local lattice clearly visible in the liberal labeling program. This lattice does not show up in the conservative labeling program since its center is labeled suspicious and, therefore, is not used to label nearby points according to its local lattice.
The second visualization of the results obtained from practicing the method of the invention is shown in
The third visualization for this same dataset is shown in
The least squares lines in this representation can be used to extend the global structure into sparse regions and enable the assignment of row and column indices to centroids that were left unlabeled by the local structure method. The grid can also be extended across empty regions to connect isolated groups of centroids.
The closeness of a centroid to the nearest grid intersection can be used as a fourth measure of the confidence in its row and column assignment. The row and column assignments of centroids that are further from an intersection of the grid may be questionable. Using this measure would enable the method to perform further consistency checks thereby resolving some ambiguities and identifying others.
All three visual representations make it easy to spot the missing rows and columns. The grid and the lattice representations make clearly visible any misalignment between two intentionally separated regions.
The sensitivity to parameter values is further illustrated by considering the labeling for the dataset from
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|Historx, Inc.
|Method and system for standardizing microscope instruments
|Aug 29, 2008
|Jul 12, 2011
|Historx, Inc.
|Automatic exposure time selection for imaging tissue
|Jan 24, 2011
|Sep 27, 2011
|Historx, Inc.
|Method and system for standardizing microscope instruments
|Jan 20, 2011
|Feb 21, 2012
|Historx, Inc.
|Method and system for standardizing microscope instruments
|Aug 7, 2008
|Feb 21, 2012
|Historx, Inc.
|Method and system for determining an optimal dilution of a reagent
|Aug 5, 2008
|Apr 17, 2012
|Historx, Inc.
|Methods and system for validating sample images for quantitative immunoassays
|May 14, 2008
|Dec 18, 2012
|Historx, Inc.
|Compartment segregation by pixel characterization using image data clustering
|Mar 21, 2012
|Apr 9, 2013
|Historx, Inc.
|Methods and system for validating sample images for quantitative immunoassays
|Dec 14, 2012
|Feb 18, 2014
|Historx, Inc.
|Compartment segregation by pixel characterization using image data clustering
|Sep 15, 2009
|Jan 19, 2016
|Novartis Ag
|Reproducible quantification of biomarker expression
|U.S. Classification
|382/288, 382/289, 702/152, 382/129
|International Classification
|Cooperative Classification
|European Classification
|Jan 23, 2003
Owner name: CHROMAVISION MEDICAL SYSTEMS, INC., CALIFORNIA
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text:
It was wine that got me into WordPress
I'm not kidding. Blame the wine for the direction my life has taken because without it I would not be a WordPress Consultant today.
In 2007, after 15 years of print writing, I launched Tasting Room Confidential on Blogspot.com - now Blogger.com - because that was the easiest way to get started. Blogger.com is like training wheels for blogging and fills a lot of needs, but eventually I grew out of it. I just wanted more. More variety, more functionality and more control.
I wanted WordPress
In early 2010 – while working on a Master Certificate of Internet Marketing through the University of San Francisco – I took the plunge into WordPress. I have never looked back. I can't say it was always easy, teaching myself the way I did, but it was always ultimately satisfying.
Learning more about WordPress made me want to do more with it. I experimented by building a site for friends, which allowed me to work with different themes and plugins.
I realized that WordPress is a lot like wine: you can never know everything because everything changes constantly. You could learn something new everyday - forever.
Designing with WordPress
In 2011, I went pro by starting this blog, Blogsite Studio, and blogging about WordPress.
I began designing sites, starting with a winery and food company.
Since then, I've created dozens of sites, and each one has taught me something new about WordPress. Here's more about my Web Design Services and samples of the Blogsites I've designed.
The biggest thing I learned when I became a WordPress site designer is that – unlike designing traditional HTML websites – WordPress is so intuitive and user friendly that people naturally want to do it themselves.
Clients don't want or need a webmaster to change every line of text or add every photo. Business people want to be able to update their own site whenever they please.
I realized what users need is someone to coach them on WordPress.
That's why I became a WordPress Consultant and Tutor
I had been teaching WordPress to some of my writer friends and discovered that I had a knack for training.
So I started a Meetup.com account called WordPress Workshop, a weekly hands-on meeting at Blenz Coffee. I'm currently taking a hiatus from WordPress Workshop, but planning to restart it later this year.
I still work privately as a WordPress Consultant with writers and business owners in WordPress tutorials, helping them to develop the techniques they need to build and manage their sites. And I love to evangelize!
I'll speak about WordPress anytime
I've spoken about WordPress at WordCamp Vancouver 2012 and WordCamp Vancouver 2013, as well as at the BC Association of Travel Writers Symposium in 2012.
In 2014, I addressed the Vancouver Business Network as well as the YVR Bloggers Meetup, evangelizing about WordPress. For a while, I was a regular on the web show, Women Talking Tech, talking about all things technology.
This year, I have so far appeared at the Blog Mastery Conference, discussing WordPress web design and at the BCATW Meetup talking about starting a travel blog. I also flew to WordCamp Las Vegas to speak about Yoast SEO.
My message: it's fun to use WordPress, you just have to know how. And, there are many ways to learn.
Ebooks are the new textbooks
In 2013, I wrote a blog series called, Create a WordPress Website to give my readers a step-by-step guide to setting up a new site. I soon released it as an ebook called, Create a WordPress Website in Ten Easy Steps. The new version, which is updated for WordPress 4.1 is available on this site, on Amazon.com and on Smashwords.
In May of 2015, I released my much anticipated follow-up ebook, Escalate Your WordPress Website: Twelve Ways to Blog at a Higher Level, which I'm offering as a free gift to new subscribers.
My Media Clips
What Can One Learn from the First WordCamp - September 25, 2015 on WPKube.com
A Tribute To Women In WordPress - March 7, 2014 on Cloudways.com
WordCamp Vancouver Speaker Mari Kane Talks About Her WordPress Journey - April 25, 2014 on Cloudways.com
Bloggers need to network too, with Mari Kane - April 24, 2015 on Fuel Radio
Get your blog on!
To me, blogging is not only a satisfying creative outlet, it's the best way to increase traffic to your website and promote yourself.
My aim is to help people and businesses achieve their internet marketing goals by creating attractive and functional blogsites with WordPress.
Let me help you make the leap into the wonderful world of WordPress.
-Mari Kane
Call me at 604-682-8973 or email me!

subset: commoncrawl


text: New Hartford Police [ Man Uses Pepper Spray On Best Buy Employees – Attempt To Identfy By Luke Austin March 14, 2014 The New Hartford Police Department is asking for your help identifying a robbery suspect. Read More Category: Utica-Rome News Tags: attempt to identify, new hartford police, robbery suspect Send to a friend! Share on Facebook Share on Twitter Pin it! Reddit This! Share on Tumblr!


text:
A programme of new forest works supported by works in the gallery, curated during the first year of Adam Sutherland's tenure as the new director. The artists and the programme were keen to distance themselves from the work that had preceded the new directorship and were generally confrontational in feel. An attempt to establish an honest response to the new conditions of Grizedale Forest as its main business shifted from timber into the recreational leisure industry.
Works included the forest billboard with an image by Marcus Coates and Calum Stirling's 'Logo Wall' - a variety of sports wear logos embedded in a dry stone wall. The programme attracted a lot of media attention and widespread criticism from local artists and visitors. It was also the blueprint under which local artists would - 5 years later - develop FRED - the annual art invasion... the sins of the fathers….
website design & build by theusefularts.org.


text:
The Burroughs' book which, with The Soft Machine and Nova Express, completes the classic Cut-Up Trilogy.
'Earth was under attack, but by whom? The Insect People of Minraud? The Nova Mob? The "White Hunters"? Representatives of Hassan i Sabbah or the White Goddess?'
'Language is the worn coin pressed silently into my hand. Inspector J. Lee of the Nova Police knew them by many names - but he only knew one way to stop them.' 'The only weapon was silence : silence to say goodbye - by silence to say good. "You see, gentlemen, what we call history is the history of the word - and the word is a killer virus!"' A prophetic vision of a world in which technology has gone haywire, The Ticket that Exploded completes Burroughs' classic Cut-Up Trilogy.
About the Author
William Burroughs was born in St Louis, Missouri in 1914. A Harvard drop-out, Burroughs lived in Mexico, Tangier and the UK, and was a heroin addict for many years. Immensely influential among the Beat writers of the 1950s - notably Jack Kerouac and Allen Ginsberg - he already had an underground reputation before the appearance of his first important book, Naked Lunch.
Originally published by the daring and influential Olympia Press (the original publishers of Henry Miller) in France in 1959, it aroused great controversy on publication and was not available in the US until 1962 and in the UK until 1964. The book was adapted for film by David Cronenberg in 1991. William Burroughs died in 1997.
'As filthy and boggling as it was in 1968 - would he get away with it today?' Time Out 'The fold-in technique gives excellent comic and satiric results...cleverly combines the raw energy of lowbrow spy- and science-fiction with the brutal unfamiliarity of hardcore pornography.' Spectator 'He has a savage sense of comedy and a cleverly educated ear for the haunting casual phrase.' Scotsman 'In Mr Burroughs's hands, writing reverts to acts of magic, as though he were making some enormous infernal encyclopaedia of all the black impulses and acts that, once made, would shut the fiends away forever.' New York Times 'His Swiftian vision of a processed, pre-packaged life, a kind of electro-chemical totalitarianism, often evokes the black laughter of hilarious horror.' Playboy Praise for William Burroughs: 'Burroughs is the greatest satirical writer since Jonathan Swift.' Jack Kerouac 'Burroughs' voice is hard, derisive, inventive, free, funny, serious, poetic, indelibly American, a voice in which one hears transistor radios and old movies and all the cliches and all the cons and all the newspapers, all the peculiar optimism, all the failure.' Joan Didion 'The only American novelist who may conceivably be possessed by genius.' Norman Mailer 'In the English language, William Burroughs is the greatest writer alive. His imagination has tackled head-on the post-war world, with its huge bureaucracies and sinister complexes. He has a paranoid vision, but as he himself said: the psychotic is someone who knows what's really going on.' J. G. Ballard, Sunday Times 'William Burroughs broadened people's conception of what makes humanity. In that way, he really was an American hero, a hero writer, and also just a great man.' Lou Reed
Audience: Professional
Format: Paperback
Language: English
Number Of Pages: 168
Published: 29th April 2010
Publisher: HarperCollins Publishers
Country of Publication: GB
Dimensions (cm): 19.700 x 13.000 x 1.4
Weight (kg): 0.128


text: The girl I'm dating is fantastic but she is about 5-6 inches shorter than I am. I enjoy sitting and looking into her eyes because that is usually the only time I see her not looking up at me while I look down on her. Otherwise I just get to see the top of her head. Should that be something that should not bugs me?


text:
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text:
I really like this guy and want to ask him out. I don't know when I'll see him next but I want to ask him when I do. He's a Freshman and I'm a Sophmore, we don't go to the same school though. How do I go about asking him?
Most Helpful Girl
Just ask him to hang out, set a day, time and meeting place. And get his number.11|00|0


text:
This looks fun and amazing! Is there any way to buy in yet? Are you planning on some sort of alpha/beta testing? Steam Greenlight maybe?
This looks fun and amazing! Is there any way to buy in yet? Are you planning on some sort of alpha/beta testing? Steam Greenlight maybe?
Love this! This is coming along beautifully.
if you code them that way it becomes more difficult to evade them, and marks for special areas with more powerful enemies, in which you have to absolutely use your surroundings to achieve perfect stealth.
Let's say for instance that you jump up from the perception zone into a hole on the ceiling (or a ceiling structure of some sort). The guard follows you until his line of sight is cut short, and then comes over to your last x position (projected to the ground) to check wheter you were a fast animal jumping up or a real foe. Then you (the player) would have to either ambush the guard from above or climb up as far as you can to stop him from alarming other guards.
But the most fun part is this: maybe your main character could have some short range teleportation ability later on (like a magic throwing knife that lets you switch positions) that can instantly snap you to the nearest walkable surface, be it walls or ground floor. If you ever encounter a enhanced guard (like the one i was just speaking about) and you jump up to avoid his line of sight he can follow you with his gaze, so you can't evade him, but then you trigger the teleportation to the ground in mid jump and you get just short of 1s of confused enemy since it couldn't follow you down due to the teleport speed. This enables you to get a preemptive attack (with some bonus added) by which then you could either flee or continue fighting.
or maybe you could just dash to an enemy near a wall and trigger this ability while he starts to swing his weapon at you: this way you could add a modifier for the direction of the teleport snap (not just nearest, but depending on controller direction) and appear crouching on the wall behind him like a ninja, ready to launch a backstab attack and maybe lash out a combo. This could be useful in boss battles to give the boss more overwhelming strength but also a way out for the player if he starts using his head to evade.
You could also combo the teleport to kill a lot of enemies in a row in an otherwise impossible situation using strategically placed platforms to propel you from point to point while slashing, like some swords manga.
There could also be an item that lets you spawn temporary platforms (they disappear very fast and cooldown time is inversely proportional to experience) to get some dynamic style fighting with high speed moves. You could also use this ability to enhance stealth to difficult levels, in which you actually have to create your stealth and mantain it in a strategic way not to be found.
Aaaah, ideas ideas. Keep this project up, it's super cool!
Last edited by max.pareschi; 07-18-2016 at 10:09 AM.
100% understood! Good luck guys, I'm sure it will be a huge success.
Hi folks,
Sorry for the lack of updates over the last few weeks. I took a little break following Develop and Radius, as well as working on a few TOP SECRET things that will be shared when the time is right
I'm currently split between two tasks - AI improvements that improve our stealth gameplay, and fleshing out the outer edges of the world map.
World Map
I've been working on making rooftop sections at the very top of the map. This is an area I'm sure will get lots of iteration, because we'd like a player who's really mastered the moveset to be able to get across here in one fluid motion.
To start with, I'm just blocking out the space, seeing what feels right, and how to sensible manage the height change between the two neighbouring sections. Currently it feels pretty good to climb the towers internally, but not so good from the outside. I'll be focusing on breaking the outline up a little and try to add some more interest as you travel between them.
While doing this, I've also started adding to my wishlist of future player controller improvements. Things like being able to interrupt some animations, and some 'feel good' things like caching of jump inputs, and allowing a little tolerance when making jumps off ledges.
Also started to prototype some of the movement features that'll be unlocked during the course of the game, but I'll save that for another update
Meanwhile, over in AI town I've been trying to work out the best way to approach doors.
The current implementation uses nav link proxies, and actually works surprisingly well - Kudos to one of our coders, Rex, for digging into these because I couldn't understand how to make them do anything useful!
However, I'm trying to take a pretty tough stance that as much decision making as possible for the AI should happen as behaviours within the behaviour tree. Currently, the use of nav link proxies breaks this rule pretty fundamentally. From the behaviour tree perspective the AI just uses 'move to' and either reaches it's final destination, using doors along the way, or doesn't make it.
I'm pushing for a solution that lets me setup lots of fall backs in the tree to allow for things like unlocking doors, breaking doors, and anything interesting we might come up with in the future. I have been heavily inspired by the examples given in this gamasutra article - http://www.gamasutra.com/blogs/ChrisSimpson/20140717/221339/Behavior_trees_for_AI_How_they_work.php
To that end, I've mocked up my own version using placeholder nodes in a behavior tree:
I have no idea how a 'sweep for doors' function would work, but isn't that what coders are for?
Well, that's all for now, I'll provide another update when some of these things are implemented. Fingers crossed it works as well as it does in my head!
In case you missed it, we have an exclusive interview with Jeremy Peel (posted on PCGamesN and the Unreal blog), covering various parts of our development process and planning. You can read it here: https://www.unrealengine.com/showcase/the-siege-and-the-sandfox-invents-the-stealthvania
BizDev (aka Business Lizard) | PR | Marketing
The Siege and the Sandfox | Twitter | Facebook | Reddit | IndieDB
Awesome work guys, this game looks beautiful. Congrats on the Unreal Dev Grant and good luck with everything
BizDev (aka Business Lizard) | PR | Marketing
The Siege and the Sandfox | Twitter | Facebook | Reddit | IndieDB
Looks like fun.
Can't wait to play it.
Make a playable demo!!
This is a frikin mazing work yall . Best looking Stealth Action Platformer I've seen in a long time. Cant wait to give a shot. The AI and Nav systems are brilliant. The control feels I have to waite and play to see. As long as those are close-ish to Ori and Meatboy, but of course different in a much more stealthy way, I smell a money train headed your way. ^_^
Lenny Gordon
80.lv did an in-depth interview with our team about building a 2D game in Unreal Engine 4. You can read the article on their website, with accompanying pretty pictures.
Last edited by OllyBennett; 11-16-2016 at 06:52 AM.
BizDev (aka Business Lizard) | PR | Marketing
The Siege and the Sandfox | Twitter | Facebook | Reddit | IndieDB
Hello, this is a awesome project.
I love that waterfall, could you share how you create it?
I then simply layered a few particle effects on top such as slow moving squares for the water breaking the crest of the fall, another for the water motes and ribbons crashing down the waterfall to aid sense of direction and speed, a distortion particle effect to give the water a sense of refraction and finally an emitter to represent the water crashing into the surface below. It's just squares in that example but I have since gone and refined it to use another flipbook to help represent the water being churned about a bit better. Hope this helps, I know its a bit "hand wavy and ta-da!" but if you need more detail let me know and I will try.
Hello ^^
I'm a young game developer, working on my first biggish 2d game. I would love to see more tips and tuts from You since Your game looks really interesting(dope <3). Just quick question: do You have any information about paper2d plugin development? This question might seems to be out of place but since You are pushing ue4 2d boundaries, maybe You are chatting with guys from Epic Games aswell I just hope it's not dead, the plugin.
How were you able to create the game in an rpg style even though there isn't and rpg2d style option in the main screen?
Sorry Pilotflyer, can you rephrase your question at all, I'm not completely sure I understand what you are asking? Are you referring to the templates/starter content that comes with Unreal 4? If so, the short answer is we don't use any of that, we started with a blank project and built from the ground up, no starter content or templates here![/QUOTE]
Yes. I meant how did you build the game from the ground up without the starter content and the default templates?
Last edited by Boxy; 03-10-2017 at 02:22 PM.
Hello. Are You setting flipbooks for actors in behavioral trees or is it done in c++? I'm curious how to control flipbooks in 2d game in ai system without coding and is it posible. Couldn't find any info about that in documentation or in forum threads.
We're not directly setting anything for the flipbooks via the behaviour trees. Our coder, Rex, built the animation system completely separately to the AI. This system is shared by all characters (player and AI) and the Behavior tree just makes calls to this.
This system was initially prototyped in blueprints, with the core functionality being moved to c++ over time. However, there's nothing really stopping you doing it all in BP if you wish, just moving to C++ suited our specific use case. However, I'd suggest that directly manipulating flipbooks in a behaviour tree probably isn't the best way to go.
Hey everyone, something else I forgot to mention, I'm now doing regular livestreaming of development over on Twitch.
Currently my schedule is Tuesday's 1800 BST / 1300 EST + Thursdays 2000 BST / 1500 EST. The general idea is to stream either before or after Epic's own streams. You can check that out here - https://www.twitch.tv/crwilso/
some eye candy pixel art here, good job
Marketplace Assets: Neon Pool Table - Modular Footbridge - 10 Stylish Fantasy Weapon - Shop Signboard - Badminton - Homemade Weapon
Marketplace WIP:Modular Scifi Experimental Lab
Folio and Others: - Portfolio - Youtube
We've posted a pixel art animation dev blog at our new location on the Chucklefish forums. You can read it here.
It details our development of our talking sprites, from almost static with just mouth motion, to animated bodies with gestures. Give it a read and let us know your thoughts.
BizDev (aka Business Lizard) | PR | Marketing
The Siege and the Sandfox | Twitter | Facebook | Reddit | IndieDB
Just as a reminder to any of those using Blueprints for AI or wanting to look over some of our processes for making a 2D game in Unreal Engine 4, we have a number of dev blogs and dev videos available to view at our subforum on the Chucklefish site here.
We're very much open to your suggestions, advice, and feedback too, so please let us know if you have any you would like to share. Thanks.
BizDev (aka Business Lizard) | PR | Marketing
The Siege and the Sandfox | Twitter | Facebook | Reddit | IndieDB
Awesome work on your project! You can see the care put into every aspect of it. I really like the implementation of the tutorials into the game so it doesn't break the flow of the game. Too many games already have that "HEY STOP!!!! THIS IS THE TUTORIAL!" system built into it that just gets annoying after a min or two. Keep up the good work.
Hey Chucklefish!
I just wanted to reach out with one question and ask if you could detail your biggest challenge(s) working with Paper2D, particularly anything that was an issue because it's still "experimental." I'm asking because my team and I are having a hard time committing to Paper2D with our project, seeing how a lot of features aren't supported yet. We have some odd features we'd like to implement in 2D (i.e. isometric perspective, dynamic lighting) and we've had a big uphill battle finding solutions from the get-go. That said, it looks like Epic is keen on supporting Paper2D, so we're hopeful that it'll catch up with the rest of the engine!
Thanks for any comments/insights you have, and take care,
Pixel art here is totally eye candy, I love it so much
Marketplace Assets: Neon Pool Table - Modular Footbridge - 10 Stylish Fantasy Weapon - Shop Signboard - Badminton - Homemade Weapon
Marketplace WIP:Modular Scifi Experimental Lab
Folio and Others: - Portfolio - Youtube
The other main challenge we have had, on and off, is the way lighting works in conjunction with an orthographic camera. Currently we have migrated to the forward renderer rather than the deferred renderer, as lights do not display in deferred when viewed through an ortho camera. We hope to look into this in the future and maybe get back to the deferred renderer, maybe.
Other than these challenges, I can honestly say working with Paper2D has been fairly easy for us, its great and not enough people give it the credit it deserves. It does so much that you would need to get multiple plugins and marketplace assets to do in other game engines.
As for dynamic lighting, again, it should work 'out the box'. Drop your lights into the world and they should light any surface they hit, as long as your material is setup to be lit (default epic ones are unlit). If you are looking for dynamic shadowing, such as in the awesome looking Pathway, you are probably going to need some code support to sort that out if you use an ortho camera. I believe you only get shadowing from the directional light at the min when using ortho cameras, if you go with a perspective camera you should get all dynamic shadowing but it might not be the right camera for your game. I'd say if you want an isometric view you don't want a perspective camera, but anything can work.
Hope this has helped you, if you got any other questions or need more clarity, let me know and I'll try my best but I'm just a lowly pixel pusher.
Hey Boxy, thanks for getting back to me! Definitely appreciate the input and help - I hadn't heard of Pathway and am blow away at how good those shadows look! It's honestly inspiring to see games like yours and Pathway with such a high level of polish. I'm also excited to hear that you've found Paper2D to work well for you and your team. My team and I love using Unreal and are more than happy to have the chance to stay in our comfort zone in terms of engine and tools.
Funnily enough, we were able to resolve a lot of our problems in the past 24 hours in regards to isometric perspective, layer rendering, and lighting. While getting the isometric TileMap set up wasn't a challenge, we were having problems with layer rendering and lighting in and isometric TileMap. As you mentioned, migrating from deferred to forward rendering was a key first step. We also played back and forth with ortho and perspective cameras and found the former to be the solution we needed if we were to go down the isometric route. That all said, with some code support we now have our rendering and lighting working properly with the different layers in our TileMaps, and are excited to see how well this system works as we expand. We haven't gotten our feet wet with 2D animations in Unreal yet, but I'm hoping Flipbook is as intuitive as other 2D animation systems we've used, such as Unity's.
Anyways, thanks again for the wise words and quick response!


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The apprenticeship is an excellent start for someone looking to gain grounding in professional administration support work.
We are looking to recruit an Apprentice within our Revenues & Benefits Team based in Bideford.
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This general LLM programme is the ideal choice for students wishing to expose themselves to diverse areas of advanced legal scholarship. Edinburgh Law School offers a very wi...


text:
Monday, April 21, 2008
Manitoba Featured in PC World
Wednesday, March 12, 2008
Video Games Live press release
Video Games Live
Presented by Future Shop
Tickets on Sale Now.
Date: Thursday, June 5 Time: 8:00pm
Pre Show Festival in lobby: 5:30pm
Venue: Centennial Concert Hall
Tickets: $35.00 – 65.00 plus fees & GST
Ticketmaster at www.ticketmaster.ca or 204.780.3333
Game on! Video Games Live visits Winnipeg with a new show for 2008 featuring members of the Winnipeg Symphony Orchestra. It's the power and emotion of a symphony mixed with the excitement and energy of a rock concert and the technology and interactivity of a video game all completely synchronized to amazing cutting edge video screen visuals, state of the art lighting and special on-stage interactive segments with the audience
Video Games Live is a multimedia concert experience featuring the best music and exclusive synchronized video clips from the most popular games from the beginning of video gaming to the present. Performed by top orchestras and choirs, the show combines
video clips and specially designed Laser and light sequences, special FX showcasing over 50 different games in a wide variety of styles. The show has a segment dedicated to the old arcade classics – but there is also modern music and older, more memorable pieces that few have ever heard live.
Tommy Tallarico and Jack Wall, the creators and producers of the show are actual video game composers and are personally involved in every show. Tommy is the charismatic host and Jack is on stage as the conductor. The video game industry takes this opportunity to help encourage and support the culture and art that the video game world has created.
Video Games Live had a fabulous 2007 selling out venues all over the world, appearing on Spike TV's Video Games Award Show, opening the LA Live Nokia Theatre and a worldwide recording deal announcement with EMI Classics.
There is a pre-show festival & post show meet and greet (with industry designers, creators, voice actors, performers and special guests) open to all ticket holders – at NO extra cost. Pre-show activities include hands-on game demonstrations, Guitar Hero competition, costume contest and prizes including a draw for a gaming console from Future Shop.
Keystone music with support from Destination Winnipeg is presenting this unique feast for the senses.
As reviewer Leo Cotterell noted, "Nothing could have prepared me for the spectacle…I found myself surrounded by video game characters. These were not paid professionals but enthusiastic fans… I discovered and enjoyed being part of another world for a night."
For more information, photos or interviews please contact:
Ruth O'Doherty, Publicist, Keystone Music Inc.
403.355.2361 firstname.lastname@example.com
Bryan Taylor, Director of Events and Operations, Keystone Music Inc.
403.355.2349 email@example.com
Thursday, March 6, 2008
Business Plan course for new media companies
For more information on the course can be found here. If you are interested in registering, click here. Questions... contact Kevin Hnatiuk at (204) 228-3252 or firstname.lastname@example.com.
This will be an impressive course to be sure!!!!
Thursday, February 14, 2008
Winnipeg Free Press article
Tuesday, February 5, 2008
CBC.ca article on Fortune Cat
Thursday, January 31, 2008
GDC or bust!!!
Fortune Cat Games Studio is going to San Francisco from February 18 - 22, 2008 to participate in the Games Developer Conference (GDC). The GDC is the largest interactive digital gaming conference in North America. The expected attendance according to the GDC website is over 16,000 people.
As a part of the "Game on Manitoba" initiative, Fortune Cat will be joining many other Manitoba and Canadian based organizations at the Canada Games Booth (#6619 - North Hall). We hope you take the time to stop by and meet with us.
More information on the GDC can be found at www.gdconf.com.
Smart Award Winners...
More information on the awards can be found at www.smartwinnipeg.mb.ca.


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Monstrance with a glass display case and lapis lazuli stones.
Made with gold-plated brass and silver. The silver shrine is surrounded by loaves ,fish, and fire (lamp).
This monstrance is handmade by an Italian crafts company that is highly experienced in liturgical furnishing. All materials are exclusively of Italian origin.
Each item is handcrafted and embellished by master Italian craftsmen. The special features of this item are the high quality materials, original designs, and delicately handcrafted construction.
- Model: OS000052
- Shipping Weight: 0.5Kg
- Materials: Silver or gold-plated brass, glass
- Origin: Italy
- MEASURESUnits:
- Height: 83.5 cm
- Inner diameter: 20.5 cm
- MEASURESUnits:
- Height: 32.87 in
- Inner diameter: 8.07 in


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KC's finest...a little jumpy?
KC police fire at backfiring van - KansasCity.com
Two Kansas City police officers who thought they were being shot at from inside a van returned fire Thursday night.
Only later did police realize that the van was actually backfiring and the man inside was not armed. He was not injured by the shots fired by police.
Windows of the police car were apparently shot out by the officers as they exited the patrol car. [say what?]
The officers were dispatched on a report of shots being fired from a white van just before 6 p.m. Thursday on Gregory Boulevard near Interstate 435.
When the officers got to the area they saw a white van parked on Gregory and pulled up near it. As they were getting out of the patrol car they heard the backfiring and fired their weapons. Police are continuing to investigate the incident.
One hopes LEOs are a little less prone to use lethal force in a situation like this. What would probably have happened to the cops if the innocent driver or a bystander had been killed by the cops? Put another way, what would happen to Joe Sixpack who started firing his legally-concealed handgun at a van that backfired because he thought he was being shot at?
That is a pretty good example of police over-reaction. If you or i did that we would be off to jail for awhile. The cops will get a pass.
Oh, that's good. A backfiring van? Classic.
You can't discern between a vehicle backfire and a gun shot?? WTF
They were probably pumping themselves up for a potential shoot out all the way to the scene!!:D
thats why the army has a day in basic training when live ammo is fired over your head.
unfortunately the officers will not be held to the same standard as civilians. if that was a regular joe those same cops would be cooking up a story about attempted murder by gun crazed civilian...
Lets see, Citizen reports of shots fired from the van. They approach the van and it backfires. They open fire. A little jumpy? Sure. When people shoot at you, you get a little jumpy. All who want to Monday morning quarterback the officers need to take a step back. In my experience a backfire can sound more like a gun shot than an actual gunshot. Think about the physics involved in a backfire. It is cause by a propellant being ignited in a tube. It is pretty much the same thing as a gun shot.
Officers that wait to see what is really going on are listed on the LE Memorial Wall in DC.
In all fairness, shooting out your own car windows in the "gun fight"? That is a little had to explain.
i bet the guy in the van will get a tuneup now:D
this azzhole was prob driving around making the van backfire...we have all done it...find a victum,while driving,get your RPM's up,turn key to off position,pump gas pedal a few times,turn key to on position then ''BOOM!''
after reading this i dont think i would call backfireing @ people funny anymore..:oi have had it done to me,while going to the mail box...it sucks!
the police have a hard enough job,i hope this guy goes and buys a lottery ticket...:rolleyes:
check this out...
Being from the KC metro myself, I posted this on our local forum.
might as well have pissed into a hornets nest.......
Epic F-up by KCMO PD. - KSCCW.com Forum
|All times are GMT. The time now is 11:26 PM.
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Thank God It's Monday
There are few reasons to look forward to Mondays and the workweek they begin...unless, of course, you frequently find free CDs and beer in your office mailbox.
Well, The Grape (2808 Greenville Ave.) and Dallas winery (yes, you read that right) Times Ten Cellars have given local drinkers at least one thing to look forward to on Mondays: an extra hour of Happy Hour prices. The Grape offers Times Ten Cellars' Sauvignon Blanc and Libertine blend for $5 a glass during Happy Hour 4:30 p.m. to 8 p.m. on Happy Mondays, and during Happy Hour from 4:30 to 7 p.m. Tuesdays through Fridays. That's in addition to other drink specials such as $2 domestics, $3 wells and appetizers in the $3 to $6 range.
Reason enough to bail from work early and have a glass of hometown wine instead.
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Semi-sheer pleated sweater cape with high-low flutter hem.63% Cotton, 22% Nylon, 15% LinenHand Wash*Import
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Job Highlights
Delhi University Recruitment 2015 Notification for 463 Faculty Posts
Delhi University Recruitment 2015 Notification was released through Advt. No. Estab.IV/256/2015. University of Delhi has released this Notification to recruit 463 Professor, Assistant & Associate Professor Posts. The interested eligible candidates may apply for these posts through online on or before the last date. This is a good opportunity for the candidates searching for Latest Teaching Jobs in Delhi. The official website to apply online for faculty posts in Delhi university is www.du.ac.in. You can apply through online form 13th May 2015 for various posts in university of Delhi.
Delhi University is one of the central university that was established in the year 1922. It is one of the top ranking university in India. Now it has released the notification for filling 463 Faculty posts. Candidates interested to apply for these posts should apply before completion of the last date. The last date for applying Faculty posts released under Delhi University Recruitment 2015 Notification is 03/06/2015. Please read the complete detailed notification before applying online. Make sure that you have met all the eligibility conditions as mentioned in the University of Delhi Recruitment Notification.
University of Delhi Recruitment vacancy Details
University of Delhi has released this recruitment notification to fill 463 Professor, Assistant & Associate Professor posts. Here we are giving the number of posts for various posts. The below table illustrates the post wise vacancy details.
|Name of the Post
|No.of Vacancies
|Associate Professor
|Assistant Professor
Delhi University Recruitment Eligibility Criteria
The Candidates applying for Various Faculty Posts in University of Delhi should met the below eligibility criteria such as educational qualification and Age limit. The eligibility criteria given in our website is based on the Eligibility criteria as given in Delhi University Recruitment Notification.
Educational Qualification:
For Professor : Candidate should possess Ph.D degree in relevant discipline.
For Assistant Professor& Associate Professor: Candidate should possess Ph.D degree with good academic record & experience in relevant field.
Delhi University Recruitment Details
Name of the Organization:University of Delhi.
No.of Vacancies: 463.
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For Assistant Professor:
- General: Rs. 300/-
- SC/ ST/ PWD candidates: Rs. 150/-
Selection Process: Candidates will be selected based on interview.
Important Dates
|Date of commencement of submission of online application
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Log on to the website www.du.ac.in.
Click on Jobs and Opportunities.
Click on Teaching Positions.
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For more details please refer official Delhi University Recruitment 2015 Notification


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Tuesday night's debate between 30th Legislative District candidates Roger Flygare and Linda Kochmar demonstrated clear differences between the two.
Kochmar, despite claiming that she would make tough choices, repeatedly said she would abide by the will of the voters on a wide range of issues. Apparently, she is unwilling to take principled positions on thorny issues, so one must ask why she wishes to represent us.
Flygare did not equivocate. His positions were clearly laid out.
Kochmar's willingness to defer issues of rights to a popular vote is particularly troubling. Had the electorate been asked, we might not have ended slavery, extended women the vote nor recognized the right to vote for people who owned no property. Our Declaration of Independence recognizes rights as being "inalienable," meaning that they cannot be taken away. Kochmar would compromise that central tenet of modern democracy that preserves the rights of the minority.
We are confronted with two assaults on civil rights: a nationwide Republican-led attack on the constitutionally guaranteed right of privacy which mandates women's choice, and an assault on marriage equality. In both cases Kochmar said she would kick the can down the road (bow to the will of the people). In sharp contrast, Flygare voiced unequivocal support for those rights.
Kochmar's weasely responses to issues of civil rights surely ought to disqualify her for the state Legislature.


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I've noticed this a lot actually and they will put girls like us in embarrassing situation because we will get judged too just because some of them are like that. I've never seen "attractive" girl complaining about it
Most Helpful Guy
There are pros and cons to being a woman but the pros are correlated to how physically appealing you are.
Ugly women have all of the cons of being a woman without any of the pros. Unsurprisingly they are not happy and have the most incentive to change gender dynamics.
Im not saying there aren't any beautiful feminists because there are (ex: Emma Watson). However almost all of the agressive, screaming, rioting in the streets sort of feminists are conventionally unnatractive.11|00|0
Not happy about what
Most Helpful Girl
of course... Unattractive girls. I never heard something so idiot, do you know that a lot of feminist don't wear make up, don't wear dress / high heels and don't shave their legs? You can"t call them attractive unless you uglier than that. That I don't wish cause you ll be really repuslsive00|00|0


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And, of course, the fine print:
Get free shipping on your next order when you spend over $100. The discount will be automatically applied to your purchase; no promo code required. The subtotal of your purchase must be $100.01 or greater after any promotional discounts have been applied. Additional fees such as tax or shipping and handling do not apply toward your minimum subtotal requirement of $100.01. Offer applies to standard shipping only. Certain items that require additional shipping and handling are excluded from this promotion and do not apply towards your $100.01 subtotal minimum. Offer subject to change.
Architectural by design, this geometric pendant necklace is inspired by the washed out shapes and sun-bleached forms of the desert. Bits of wire are carefully sawed, curved and soldered precisely into place to create the vertically stacked or scalloped bib design for this necklace, and the openwork pendant is then cast in solid sterling silver. Each one is polished and tumbled for strength and shine, and for the gold variations, they're plated with 18k gold for a warm golden hue.
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This product ships from the maker within 14 days, via USPS
If you're not happy with this item for any reason, you may return it for a full refund within 14 days.


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This item from Jon Berg Fine Arts and More is now SOLD.
More from Jon Berg Fine Arts and More
Late 18th or early 19th century painting on porcelain after Dutch Old Master artist Gabriel Metsu
An antique (late 1700's or early 1800's) hand painted work on porcelain, 5 1/2" by 4 1/2" (8" by 7" in later wood frame), the subject a seated young woman in bonnet reading a letter. On the reverse in very old ink and hand is written in French "Jeune femme lisant d'apres G. Metsu" and apparent indecipherable signature/initials after that. The French translates as "Young woman reading, after G. Metsu". Gabriel Metsu (1629-1667) was a Dutch Old Master, some of whose works have exceeded one million dollars at auction. He was known for genre scenes, portraits and still life. His works took after Terborch and de Hooch. The charming piece has survived the centuries in very good condition. The frame has chipped upper corners.
If you were the pending buyer of this item, go to My Account to view, track and check payment for this item.
Jon Berg Fine Arts and More
Jon Berg
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Exactly what do males fear most about aging? Based on a current survey in the United kingdom, balding outperform impotence, weight problems, as well as cardiovascular disease for that top place. An astounding 94 percent of participants stated that losing their head of hair afraid them greater than every other age-related option. Knowing that, listed here are five advantages of hair alternative therapy.
Forget About Bald Jokes
Like a society, we have become much more open to individuals variations recently. Its no more considered allowable to poker fun at someone according to how old they are, gender, or looks. The best exception is that if that individual has lost or perhaps is losing their head of hair. Bald people are potentially the only real group that may be exposed to public ridicule with impunity. This is also true once the butt from the joke is really a guy. The main reason? Since most men eventually lose their locks, theyre expected to conquer it. Coping with losing is created even more complicated when buddies and other people deride them regularly.
Reinstates Self-Confidence
Numerous research has proven that many men lose confidence once they start losing their macho mane. As the jokes certainly dont help, the primary problem appears to become an interior one. The truth is that many males dont consider bald to become beautiful. Then when they begin lower that lonely path, it can be hard to allow them to accept their new reality.
Increases Attractiveness
It might not be an offer-breaker for many women, however, many shallow gals simply wont date a man having a bare mind. Males whore showing scalp are comfortable with this fact, also it has a tendency to affect their confidence negatively using the ladies. Trading in hair alternative therapy might help have them in the saddle again.
Turns Back the time
Be it true or otherwise, most people connect hair loss with senior years. Consequently, even more youthful men whore hair deprived are frequently viewed as searching over the age of they are really. Although this belief harm your self-confidence, but this may also lead him to appear less energetic and capable. This could hurt him in present day ultra-competitive place of work, where youth and are valued most importantly else.
Stops the Charade
For the reasons we have pointed out, some men attempt to hide their head of hair loss. They may put on lots of hats, employ the infamous comb-over, or stay inside on Saturday evening. But none of them of individuals tactics stop their locks from receding. The only real efficient way to handle the issue is to determine a professional.


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1. No backing out
2. Both are healthy adults
3. Both know each others presence
Advantages:Speed, Intelligence, Bite, Claws
Disadvantages: Reckless, little stratgey
Stegosaurus: Deadly tail spikes, Plates for intmidation, More stable
Disadvantages: Speed, Plates are easily injured, Intelligence
The Verdict: Stegosaurus 60-40 Allosaurus. I believe the Stegosaurus is more likley to win due to the fact if it gets lucky it can kill the Allosaurus in one clean hit.
The Battle: The Allosaurus spots a drinking Stegosaurus. He steps forwars, ready to pounce. As he heads towards his prey he recklessly charges, only he is headed from behind. He just ducks the tail swing of the Stegosaurus. He then leaps at the Stegosaurus but is forced to land hard to the ground when the Stegosaurus atempts to kill it. Soon the Allosaurus begins pacing around its prey, just out of reach of its tail. He then decideds to go for thevulerable head. But the Stegosaurus is ready swinging its tail, warding off the predator.Soon the Allosaurus charges again, slashing the Stegosaurus in the chest. Only to be grazed by the herbivores' tail. The Allosaurus' skin rips and he feels blood gush out. The Allosaurus soon begins angrily slashing the Stegosaurus flank. It soon heads towards the head and
begins to bite down and prepares to twist the Stegosaurs' neck. In Desperation the Stegosaurs jerks his bleeding head from the jaws of the predator.Then Stegosaurus then lands that one vital blow. Foot long spikes pierce and rip into the Allosaurus' flesh, only stoppoed at the ribs. But it is not enough. The Allosaurus then prepares to charge the Stegosaurus, head first. Time seems to slow down as the Allosaurus charges and the Stegosaurus swings its tail. The Allosaurus the realizes its mistake. Its eyes turn in horror to its would be preys tail. The spikes connect with Allosaurus head, piercing it in its brain and other organs in its head. The Allosaurus roars in agony as the Stegosaurus beings wrenching its tail spikes in the Allosaurs head. The Allosaurus roars again, then falls to the ground. Dead. The Stegosaurs, bloody and wounded looks at his fallen attacker, then walks away.


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Tag Archives: Adventures in Odyssey
Adventures in Odyssey
I can hardly believe that it has been over 20 years since I first started listening to Adventures in Odyssey. I listened to several radio programs regularly. Most of them from the radio station in Zarapath, New Jersey. What was … Continue reading
Posted in Introspection Tagged Adventures in Odyssey, radio, Stories of Great Christians, Unshackled Leave a comment


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Have been dating for more than a year, We have a perfect and great relationship but one thing my boyfriend always scolds me in vulgar words that really wound my heart.
To make things worst he would stop his car in public at a bus stop in town just to let go his anger by beating me,pulling my hair,kicking me and just making me cry.
I think my boyfriend is from a educated family but he was never taught to respect a girl I guess,but when it comes to love and care his great.
Most Helpful Girl
GET OUT BEFORE HE BECOMES UR HUSBAND AND You HAVE KIDS!
Darlng, that is not LOVE. If you think it is then your wrong. Please please I am begging you to remove yourself from the situation. I was with a guy like that...well not as bad, but similar. He would spit out every cuss word in the book to me. EVen curse my mom& dad to me. He also raised his hand but the third time he did I walked out and NEVER looked back. Yes, I miss him and he misses me too and he says he is sorry, but "sorry" is not enough! SO many other guys will care and love you. So many other men will want to be your everything and give you the life you really deserve. Don't be afraid to be alone for a couple of months or even a yr. I promise you will find someone better than the slimebag your dating. AS of now just distant yourself, don't "Sext" him if you or if you have delete all pics and whatever. Slowly ignore him even if he is crying and begging for him. And then completely cut him off.10|10|0


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Keep Kids Safe This Summer with Pool Safety Rules
Jun 21, 2017
When temperatures rise, we often find ourselves seeking a spot by, or in, the water. And that's why it's important to make sure our children ...
Rapides Regional Medical Center is an award-winning hospital and ER in Alexandria, Louisiana that offers a wide array of services ranging from Cancer Care to Wound Care, and our medical staff includes doctors in more than 30 medical specialties. View our average ER wait times above.
In 2012 the Alexandria hospital became the first verified Level II Trauma Center in Louisiana. In addition, we operate the area's first Certified Stroke Center, offering comprehensive stroke care from the minute a patient walks through our door to the time they go home.
The Rapides Parish hospital is also a regular recipient of the Louisiana Quality Award, which honors businesses and organizations that are leaders in the pursuit of quality and service. Rapides Regional Medical Center was also named one of the nation's "100 Top Hospitals - Benchmarks for Success." The annual study lists hospitals based on quality, operations and financial management.
211 4th St
Alexandria, LA 71301
Jul 12, 2017
Rapides Women's and Children's Hospital is adding a new class for moms this August – and this one is specifically for breastfeeding moms returning...
Jun 21, 2017
When temperatures rise, we often find ourselves seeking a spot by, or in, the water. And that's why it's important to make sure our children ...
May 31, 2017
Award given by American Heart Association/American Stroke Association, Get with the Guidelines.
May 02, 2017
Rapides Regional Medical Center provides free screenings and info for attendees
Apr 19, 2017
Free screening by local dermatologists, staff.