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|Publication number
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|Publication date
|Jun 15, 2004
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|Jun 25, 2002
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|Aug 6, 1999
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|Richard Levenson, Peter J. Miller
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|Cambridge Research And Instrumentation, Inc.
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|Patent Citations (25), Non-Patent Citations (10), Referenced by (96), Classifications (40), Legal Events (10)
|External Links: USPTO, USPTO Assignment, Espacenet
This application claims priority to, and is a continuation-in-part of, U.S. utility patent application Ser. No. 09/921,040, entitled "Spectral Imaging System" by Peter J. Miller and C. Hoyt, and filed Aug. 2, 2001 now U.S. Pat. No. 6,690,466, which is a continuation of U.S. utility application Ser. No. 09/633,417, entitled "Spectral Imaging System" by Peter J. Miller and C. Hoyt, and filed Aug. 7, 2000 now U.S. Pat. No. 6,373,568, which claimed priority to provisional application Ser. No. 60/147,636, entitled "Spectral Imaging System" by Peter J. Miller and C. Hoyt, and filed Aug. 6, 1999. This application also claims priority from U.S. provisional patent application Serial No. 60/300,696, entitled "Spectral Imaging Systems and Methods" by Richard Levenson, and filed Jun. 25, 2001. The contents of these utility and provisional patent applications are incorporated herein by reference.
Advances in the molecular understanding of many diseases and conditions depend, to a large extent, on microscopic evaluation of tissues. For example, malignant cells are identified by evaluating tissue samples. It is one of the most arduous and time-consuming tasks in pathology. To facilitate the evaluation, tissue samples can be treated with stains to provide feature contrast.
Human color vision is a form of spectral imaging by which we determine the intensity and proportion of wavelengths present in our environment, and their spatial distribution. However, unlike the eye, which breaks up the light content of an image into red, green, and blue, instrument-assisted spectral imaging can use an arbitrarily large number of wavelength-classes. Furthermore, it can extend the range to include the ultraviolet and infrared regions of the spectrum invisible to the unaided eye. The result of spectral imaging is a data set (known as a data cube) in which spectral information is present at every picture-element (pixel) of a digitally acquired image.
The invention features a method that forms one or more images of an unknown sample by illuminating the sample with a weighted spectral distribution for each image. The method analyzes the one or more resulting images and identifies target features. The identifying can include classifying regions of the images based on their spectral or/and spatial properties. In addition to classifying, the method can quantify the amount of some biomarkers either in combination or apart from the classifying. The identification of particular target features can guide the automation of subsequent processes.
In general, in one aspect, the invention features a method for imaging. The method includes: obtaining a spectral weighting function indicative of an attribute of the reference sample; illuminating a target sample with light whose spectral flux distribution corresponds to the spectral weighting function to produce a corresponding target image, wherein the target image is indicative of a response of the target sample to the corresponding illumination at multiple spatial locations of the target sample; and identifying one or more target features in the target sample based on the target image.
Embodiments of the method may include any of the following features.
The spectral weighting function may be obtained from a set of reference images. The spectral weighting function may be determined based on at least one of principal component analysis, projection pursuit, independent component analysis, convex-hull analysis, and machine learning The method may also include illuminating a reference sample at each of a plurality of pure spectral bands to produce the set of reference images, wherein each reference image is indicative of a response of the reference sample to the corresponding illumination at multiple spatial locations of the reference sample.
The method may also include determining one or more additional spectral weighting functions indicative of additional attributes of the reference sample based on the set of reference images; and illuminating a target sample with light whose spectral flux distribution corresponds to each of the additional spectral weighting functions to produce additional corresponding target images, wherein each target image is indicative of a response of the target sample to the corresponding illumination at the multiple spatial locations of the target sample. The response of the reference sample may include transmission, reflectance, or fluorescence. The response of the target sample may include transmission, reflectance, or fluorescence. The spectral weighting function may include multiple ones of the spectral bands. The spectral weighting function is a single one of the spectral bands.
The method may include preparing the reference and target samples with markers suitable for chromogenic in-situ hybridization. The response of the target sample to the spectral weighting function may correlate with the presence of the in-situ hybridization marker in the target sample. The method may include preparing the reference and target samples with a marker suitable for color immunohistochemistry. The response of the target sample to the spectral weighting function correlates with the presence of the color immunohistochemistry marker in the target sample. The method may include preparing the first reference and first target with a general stain.
The identification may include assigning the one or more target features to one or more classes. The identification may include quantifying the amount of a chromogen in a target feature.
The method may further include automating a subsequent process based on the identification of the one or more target features. The subsequent process may include laser capture microdissection. The laser capture microdissection may include directing laser energy to the identified target features in the target sample to remove corresponding portions of the target sample. The method may include performing mass spectroscopy on the portions of the target sample removed by laser capture microdissection process. The mass spectroscopy may be protein mass spectroscopy. The may also include performing protein purification on the portions of the target sample removed by laser capture microdissection process. The subsequent process may include determining the extent of a condition or disease in a target sample. The condition may be fibrosis. The condition may be chronic organ rejection
The method may also include repetitively illuminating the target sample with light whose spectral flux distribution corresponds to the spectral weighting function to record the target image as a function of time. The identifying one or more target features may be based on the time dependence of the target image.
This application incorporates other documents by reference. In case of conflict, this document controls.
The details of one or more embodiments of the invention are set forth in the accompanying drawings and the description below. Other features, objects, and advantages of the invention will be apparent from the description and drawings, and from the claims.
FIG. 1 is a flow chart of a method for using a spectral illumination system to analyze a sample.
FIG. 2 shows a spectral illumination system for spectral measurements of a sample on a microscope slide.
FIG. 3 shows a first implementation of the spectral illuminator.
FIG. 4 shows another implementation of the spectral illuminator.
FIG. 5 shows another implementation of the spectral illuminator.
FIG. 6 shows another implementation of the spectral illuminator.
FIG. 7 shows another implementation of the spectral illuminator.
FIG. 8 shows a block diagram of the electronics used in the spectral illuminator.
Like reference symbols in the various drawings indicate like elements.
The invention features a method for recording images of an unknown sample and identifying target features in the unknown image. Although the method finds use in many applications, it is particularly useful in the field of biological tissue examination.
A method 100 for identifying target features from one or more images of an unknown sample is outlined in the flowchart of FIG. 1. Referring to FIG. 1, method 100 begins by recording one or more reference image cubes from one or more reference samples (Step 110). An image cube is the spectral response of the sample as a function of spatial position and wavelength. If the goal were to assign target features to two classes (for example disease vs normal cells) then representative image cubes would be recorded for both classes. In other applications, if the goal were to quantify the amount of some biomarker such as those in used immunohistochemistry, then the reference samples would include a sample with the pure immunohistochemical biomarker. Using standard techniques, these reference image cubes form a training set. The training set is then processed to define one or more spectral weighting functions (Step 120). A spectral weighting function is the weighted sum of the spectral response at various spectral bands. The spectral weighting functions are determined to be functions whose values are particularly relevant in identifying the target features. Once the spectral weighting functions are defined the spectral illuminator illuminates the sample with a flux distribution equal to the light distribution of the spectral weighting function (Step 130) and the light intensity recorded by a detector is the spectral weighting function. The resulting images are processed to identify the target features. The identification is based on a comparison between the images of the unknown sample and the training set (Step 140). Once the target features in the sample have been identified, subsequent tasks can be automated based on the identification (Step 150).
The method is suitable for the analysis of biological tissue samples. In the following, we describe examples of techniques for preparing such biological tissue samples. To enhance the visibility of features, staining can be used to improve contrast in such tissue samples. For example, fixed slices of tissue can be stained with hematoxylin and eosin (H&E). Because these dyes have certain affinities for cellular constituents they enhance contrast and visibility of cellular features. There also other stains that are suitable for use with method 100. For example, cytopathology specimens, including cervico-vaginal "Pap" smears, can be stained with other dyes, including the Romanowsky-Giemsa formulation (azure B and eosin Y) and the Papanicolaou stain (hematoxylin, orange G eosin Y and light green). Other suitable stains include toluidine and methylene blue.
In addition to staining specific cellular features, conditions such as inflammation and neoplasia and other physiological and pathological processes affect the distribution, intensity and color of the stains. Thus the overall spectral signature of such stains is characteristic of the various cellular components and further detailed spectral analysis can diagnose subtle physiological and pathological changes. For example, by analyzing the image cubes of stained specimens, neoplastic vs. normal cells of the same cellular lineage can be distinguished and classified.
Such general stains typically bind non-specifically. For example, hematoxylin, a blue dye, stains predominantly nuclear features whereas eosin stains cytoplasmic and extracellular components pink-red. Classifying target features using such general stains is based on empirical information. The method forms a training set of known samples that have been stained with such general stains. Typically, the known samples include representative samples for each class that may later be identified.
In contrast to the general staining methods, other techniques such as histochemical staining generally involve a previously established binding mechanism. For example, histochemical stains can be directed specifically at cellular constituents such as elastin or collagen. Typically, these methods employ chemical reactions targeted at different biochemical species in tissue and generate a color signal via various mechanisms. Such techniques can be used to identify specific structures; constituents and cell types and can also be used to provide quantitative information. The method first obtains spectral information about the tinctorial properties of the various components in a sample, and uses this spectral information to identify and quantitate such components.
Another suitable technique for marking specific features with chromogens is immunohistochemistry (IHC). IHC is the detection of antigens using antibodies that are coupled to some kind of chromogenic readout system. Immunohistochemistry provides a very high level of specificity in its binding due to the highly specific interaction between antibody and antigen. For example, an suitable sample preparation using IHC includes a brown chromagen (DAB) to an immunostain that recognzies the progesterone recepter and red chromagen (Fast Red) coupled to a immunostain recognizing the estrogen receptor. The levels of the chromogenic markers used in IHC can be used to measure quantitatively the levels of estrogen- and progesterone-receptors. Similar sample methods can quantitatively measure the levels of other important biological targets such as Her2-neu expression, p53, and ki-67.
In situ hybridization (ISH) is the specific tagging of nucleic acid sequences with a chromogenic readout system. Non-fading, enzyme cytochemical detection methods for hybridization sites have been developed and have led to chromogenic in situ hybridization. Typcial ISH probes include centromeric markers, telomeric markers, subtelomeric markers, single gene markers as well as mutation, deletion, amplification and rearragement detection systems. In a fashion similar to IHC, chromogenic ISH provides highly specific information due to the specificity of the hybridization. An example of samples labeled using ISH include bladder carcinoma tissue sections hybridized with centromeric chromosome I probe coupled to biotin or dioxigenin and visualized using DAB or HPR-TMB respectively.
Combining these various techniques simultaneously can produce an especially information rich composite probe of tissues. For example, chromogenic ISH can be combined with IHC to provide multiparameter molecular characterization of cancer biology. In such settings, the method 100 is well suited to analyzing images of samples that have been treated with such composite probes.
In what follows we first describe a spectral illumination system especially suited for implementing the method. This is followed by an expanded description of method 100. Thereafter, we describe alternate embodiments of the spectrally illuminator and we describe the spectral illumination system electronics.
A system for making spectral image measurements is illustrated in FIG. 2. A spectral illumination system 60 includes a spectral illuminator 1 driven by electronics 40 and controlled by computer 63. The spectral illuminator is controlled to provide light output, I(λ), that can be independently varied for bands of wavelength for any band in the illuminator bandwidth. Typically the bandwidth is centered in the visible spectrum. The spectral illuminator is described in detail below. A spectral illuminator is suitable, so long as it provides a plurality of well-defined bands with minimal overlap that span the spectral ranges of interest. The flux from each band is adjustable and reasonably reproducible, and multiple bands can be active at one time.
The light output of the illuminator is directed into microscope 65. The optical elements 68 of the microscope guide the light toward the sample 67 and from the sample towards detector 61. The light illuminates sample 67 and the transmitted light is directed to detector 61 via lens 161. The detector collects an image of the sample. The signal from the detector is digitized by digitizer 62 and is transferred to computer 63. The control of the spectral illuminator and the processing of the resulting images are performed by instructions from software 64.
The system shown in FIG. 2 is suitable for obtaining transmission images using a microscope. In other embodiments, the system can measure reflectance, scattering, or even fluorescence. Furthermore, the system may use microscope imaging technqiues or macro-scopic imaging techniques.
In general, the signal induced at a detector by light in an infinitesimal spectral band centered at λ0 and having width dλ will be given by
where D is the signal produced at the detector, I is the illumination flux, S is the spectral response of the sample, and R is the detector responsive for the given wavelength.
S may indicate reflection, transmission or scatter, depending on how the sample is being illuminated and viewed, (i.e. by a transmissive, reflective, or scattering arrangement). In the present context, these are all equivalent so long as the wavelength of light is substantially unaltered by its interaction with the sample.
It is possible to rewrite the Eq. [1] in terms of discrete bands, provided that the bands are chosen to be sufficiently narrow that neither the illumination, sample response, nor detector response varies greatly within a given band. Then one may write the expression for the contribution from light in the j-th band as
One normalizes spectral readings against a standard such as a white card (for reflection measurements) or a transparent reference (for transmission measurements). This normalization is performed on a band-by-band basis, and typically for the standard S(λ)=1, leading to a signal of
when the standard is read. This allows one to measure the normalized sample property S(j) by taking the ratio of the detector readings when the sample is present, relative to when the standard is present, as:
It is this same method that is used to determine the image cube for a sample. One illuminates with each wavelength band in turn and observes the detector reading D(j), under both sample and standard conditions for every pixel in the image plane; the ratio yields the sample response S(j) which when measured for each pixel r in the image plane becomes the image cube S(r,j).
As was described above, the method identifies target features in images of an unknown sample (or samples) and that identification relies on the reference images. The reference images depend on the type of identification that is to be performed by the method. For example, for classification based on samples prepared with nonspecific general staining, the reference images are representative of each class that is to be identified in the image. For example, if normal cells are to be distinguished from diseased cells then representative samples of diseased and normal cells are obtained that have been stained using the protocols used for the unknown samples. From the resulting training set, the method assigns features in the images of the unknown sample based on the training set. Programmable, reliable automated staining systems are available from Ventana Medical Systems (Tucson, Ariz.).
For chromogen quantification, the reference samples provide the spectral information about the chromogen of interest. As the actual sample can be stained using a combination of the techniques described above, the quantification of a single chromogen often requires spectral information about each of the chromogens present in the sample so that the contribution of the chromogen of interest can be unmixed from the others.
For preparation of IHC based biomakers, techniques can use enzyme-based color generation systems, usually coupled with other amplification schemes. Two enzymes are commonly used for this purpose, alkaline phosphatase and horseradish peroxidase, each of which can catalyze the deposition of colored precipitates in their vicinity.
For preparation of ISH based markers, a number of suitable enzymes for ISH detection procedures are now commercially available. For example, ISH markers are available from Zymed Laboratories Inc. (South San Francisco, Calif.). The best results have been obtained with horseradish peroxidase and alkaline phosphatase. To obtain 3- or more color results, it is possible to quench residual enzyme activity from the first rounds of chromogen deposition and then come back with other enzyme-linked probes using a different set of chromogens. For example, HRP-DAB (brown), HRP-TMP (blue-green) and AP-fast red or new fuchsin can be combined. In some embodiments, catalyzed reporter deposition (CARD) using tyramide conjugates is used.
The first step of method 100 uses the spectral illumination system 1 to obtain reference image cubes from reference samples. Typically these reference samples are prepared using one or more of the techniques described above (general staining, hsitochemistry, IHC, chromogenic ISH). The reference samples are chosen based on the type of features that are to be identified. In addition similar protocols are followed for both the reference samples as will be used for the unknown sample. This ensures that variability in the spectral features due to lot-to-lot changes in the stains or other systematic errors can be minimized. Some embodiments prepare the reference samples at separate times than the unknown samples. In yet other embodiments the spectral cubes of interest are obtained from other sources such as reference sources for spectral information about stains or chromogens. Such choices are determined by the application at hand.
As described above, the spectral illuminator 1 can sequentially illuminate the reference sample(s) band by band while recording the spectral response of the sample for each band. This data set forms the image cube. The one or more image cubes of the one or more reference samples provide the raw data sets also known as the training sets that are later used in identifying target features.
Once the image cubes are obtained for the reference samples, computer 63 processes the input raw image cubes of the reference samples and derives the spectral weighting functions from the reference image cubes (Step 120).
The spectral weighting function WF and its distribution F(j) are derived from the image cubes using methods that are familiar to one skilled in the art of spectroscopy and multispectral image analysis. Given a spectral weighting function with a spectral distribution F(j) for a specified set of bands λj, the spectral index for a sample with response S(j) is given by:
The distributions F(j) are chosen such that the weighting function WF of an unknown sample is the basis for identifying the features of interest in the unknown image. For example, when the goal is to assign target features to various classes, then the spectral weighting functions are chosen to optimize class separation. Often standard statistical tools are employed to analyze the training set data and the spectral weighting function and the subsequent target feature classification (see discussion of step 140 below) uses the information derived from such statistical analysis. When the goal is to quantitatively determine the amount of a chromogen, then the spectral weighting functions are chosen such that pixel-unmixing algorithms can extract and compute the amount of chromogen present in the spectral data from the unknown sample.
Techniques to derive the spectral weighting functions are described in the prior art such as "ISIS; An Information-Efficient Spectral Imaging System", by W. C. Sweatt, C. A. Boye, S. M. Gentry, M. R. Descour, B. R. Stallard, C. L. Grotbeck, Imaging Spectrometry IV, M. R. Descour, S. S. Shen Ed., Proc. SPIE, Vol. 3438, pp. 98-106, San Diego, 1998; "Construction of filter vectors for the Information-efficient Spectral Imaging Sensor", by B. R. Stallard, , S. M. Gentry, Imaging Spectrometry IV, M. R. Descour, S. S. Shen Ed., Proc. SPIE, Vol. 3438, pp. 172-182, San Diego, 1998; "Spectrally Adaptive Light Filtering", by P. I. Shnitser, I. P. Agurok, Proc. SPIE, vol. 3140, p.117-27, 1997; "High Dimensional Feature Reduction via Projection Pursuit", by L. O. Jimenez,, D. Landgrebe, TR-ECE 96-5, School of Electrical Engineering, Purdue University, West Lafayette, Ind. 47907-1285, 1995; all of whose contents are incorporated by reference in their entirety.
One such technique is principal component analysis (PCA). PCA is a widely used tool in a broad range of disciplines in which multidimensional data are commonplace. In the context of spectral image analysis, PCA may best be viewed as a method seeking a linear transformation of the input spectral image that optimally approximates the latter in a least square sense. The individual principal components, which are the eigenvectors of the covariance matrix of the input image, are all uncorrelated and point in the directions of maximal variance of the data points. In this way, PCA achieves what is sometimes called a faithful representation of the input data. PCA is useful in computing spectral weighting function that optimizes separation between classes.
Since PCA is exclusively based on the second-order statistics contained in the covariance matrix, it will fail to detect any higher-order structure, i.e., relationships between more than just two pixels, in the input data. Traditionally, this limitation has not received much attention because input data were generally assumed to conform to some approximation to a Gaussian distribution and for the particular case of Gaussian input data it can be shown that indeed all information is captured in the covariance matrix.
Another technique, independent component analysis (ICA) uses image transformations that explicitly incorporate higher-order statistics to obtain a meaningful representation of the image data. This is a much stricter requirement than mere uncorrelatedness of the PCA components. Only in the case of Gaussian input data, the implicit assumption of PCA, uncorrelatedness of the components is already sufficient for independence. Intuitively, ICA can therefore be understood as a linear transformation that maximizes non-Gaussianity rather than variance. A useful approach is actually to apply PCA operations prior to ICA.
Another technique is projection pursuit (PP). Often PP is used to compute spectral weighting functions that are useful for spectral unmixing. In PP, a projection index is used to find projections in multivariate data that are structured in a non-random and therefore "interesting" way. Since in most applications the Gaussian distribution is considered the least interesting (and most projection indices used in PP are indeed some measure of non-Gaussianity) the ICA components are in fact special variants of PP axes.
The type of identification that is to be performed also determines the number of spectral weighting functions that are computed. As the number of classes increases, typically the number of spectral weighting functions needed to accurately classify objects increases. Similarly when quantifying a chromogen from amongst an increasing number of overlapping stains, the number of spectral weighting functions necessary to accurately unmix and quantify that chromogen increases.
Sometimes an optimum spectral weighting function may correspond to one of the pure spectral tones, see, e.g., commonly owned U.S. Provisional Application Ser. No. 60/314,367 by Paul Cronin et al., filed Aug. 23, 2001 and entitled "Multispectral Imaging Method and Apparatus," the contents of which are incorporated herein by reference.
Once the spectral weighting functions have been determined, the spectral illumination system uses the spectral weighting functions to image the unknown sample 67. The computer 63 adjusts the settings of the universal spectral illuminator to produce an illumination spectrum with the property that
That is, the illumination distribution has the shape of the spectral weighting distribution F(j), divided by the detector responsivity D(j). The spectral weighting functions can be a single band in the spectral illuminator, or they can include multiple bands.
When the sample is imaged under this controlled illumination, the total signal at the detector D is given by
The detector directly measures the value of the spectral weighting function, without need for breaking the light up into its constituent spectrum and numerically analyzing it. The result is an optical, rather than numerical, evaluation of the spectral weighting function, with enormous increase in speed and efficiency.
In spectral illuminators which use LED's, the LED current settings which realize the required illumination spectral distribution can be calculated directly from tabulated α(j) values; the proper LED current setting for band j is simply F(j)/α(j). The scale factors α(j) are found as follows. The LED output is variable, it is possible to adjust the LED current setting so as to attain a sought-for reading at the detector, which we denote as K. By doing so in all bands, and recording the drive current settings C(j) required to meet this condition, one can calculate a calibration table of scale factors α(j):
The scale factor α(j) is the detector response per unit of LED drive current in band j.
The spectral illuminator is then set to produce the spectral distribution specified by the spectral weighting function, and an image of the entire sample is acquired. If several spectral measures are desired, then several exposures are taken. However, only one exposure is required for each spectral measure. Collecting the spectral weighting functions using this method is also known as matched filtering
In some embodiments, the spectral illumination system will record multiple images of the spectral weighting functions as functions of time. Such a time series of data allows the calculation of the time evolution of the samples spectral properties.
Having collected the values of the spectral weighting functions from the sample, the images are processed and the target features are identified (Step 140). In the first step of the image processing, objects contained in the image scene are separated from the background (image segmentation). The method segments the image by locating the target features and removing the background. In some embodiments, the image is not segmented and each pixel in the image is classified or quantified. In addition to using spectral information for segmentation, spatial information can also be used.
Once the background is removed and the images have been segmented, the target features are identified. When assigning objects into classes, the method assigns the objects into classes based on the value of one or more of their spectral weighting function. As described above, such an assignment relation typically is based on a statistical analysis of the training set data.
There are several approaches to classifying pixels in a spectral image. The minimum squared error method compares each pixel in the image with a set of reference spectra using a least-squares criterion. Other approaches convert spectra into n-dimensional vectors, and the angles between such vectors can be used as measures of similarity. Pixels are classified according to which pure spectrum they are most similar to and can be pseudo-colored to indicate the results of classification. There are more advanced distance measures, such as Mahalanobis distance, which can take into account variations of signatures. Determining which spectra to use for the classification procedure is not always straightforward. In simple cases, the reference spectra can be selected from obvious structures in the image (foci of cancer vs. normal cells, for example) or from established spectral libraries. Alternatively, informative spectra can be extracted using statistical analysis methods, such as principal component analysis (PCA) or clustering methods. Instead of using a classified pseudo-color display, spectral similarity can also be illustrated by mapping the degree of similarity using gray-scale intensity.
In an exploratory context, the target classes as well as the features used for their subsequent discrimination have to be defined initially, typically in an iterative process: first, a set of preliminary target classes is defined, either manually under the guidance of an experienced pathologist or automatically by applying a clustering algorithm to the image data; second, the features that optimally discriminate between the target classes are selected; and third, the performance of the resulting classification system is evaluated by applying it to a new set of sample image data. If necessary, these three steps are repeated with different sets of preliminary target classes until satisfactory results are obtained.
The spectral classification methods are suitable for images in which no pure spectral components are likely to exist, such as in samples prepared with general stains. In other types of images, multiple distinct spectral signals may combine to form the detected signal. Spectrally mixed pixels result when objects cannot be resolved either at an object boundary (spatial mixture), or when more than one object is located along the optical path (depth mixture). A linear combinations algorithm can be used to unmix the signals arising from the pure spectral components. Given an appropriate set of standards, the algorithm can determine quantitatively the absolute amount of each label present. The linear combinations algorithm assumes each pixel is made up of a combination of pure spectra. Typically such a set of standards is provided by the training set.
Sx,y is a vector containing the spectrum at a pixel and L is a matrix with n pure spectra and m spectral channels. We solve the equation for w, which represents the contribution of each pure spectrum in L to a pixel sx,y in the image. The L matrix is overconstrained because there are usually more spectral channels than pure spectra. We must therefore find an approximation of w. Using the least squares approximation, we minimize the residuals e to give the best w.
We can determine quantitatively how much of each pure spectrum is in each pixel in the image using w. A grayscale image c is created for each pure spectrum in the library. Each pixel in each c image is assigned a fraction, according to w, of the total intensity of the corresponding pixel in the spectral image. We first normalize the weights w to w′ so that the sum of all elements of w′=1. Let Ix,y be the integrated density of the pixel at x,y. For each pure spectrum i=1 . . . n:
The linear combinations algorithm can only be applied in situations where the pure spectra combine linearly. This property holds for fluorescence images, but transmission and reflectance images must be converted to optical density before applying the linear combinations algorithm. Once the spectral unmixing algorithm determines the amounts of the pure spectra, the amount of the chromogen associated with the pure spectrum is computed.
In some embodiments, the analysis for identifying target features may also include morphology-based analysis. In order to provide such identification, the training sets are provided with representative images of the morphological types that are to be identified.
In yet other embodiments, both spectral and spatial image information are used to identify and classify the images. For example, an algorithm known as GENIE, a machine learning algorithm, uses both spectral and spatial information (See "GENIE: A Hybrid Genetic Algorithm for Feature Classification in Multi-Spectral Images", Simon Perkins, James Theiler, Steven P. Brumby, Neal R. Harvey, Reid B. Porter, John J. Szymanski, and Jeffrey J. Bloch Proc. SPIE 4120, pp 52-62, 2000). Other embodiments can use other machine learning algorithms or genetic learning algorithms.
With the results of feature identification subsequent tasks that depend on the outcome of that identification can be automated (Step 150). One such task is the automation of standard laser capture microdissection (LCM). Standard LCM is a method by which pure cell populations are selected by an expert operator from a heterogeneous sample that may contain many types of cells in addition to the target cell type. In the case of the PixCell LCM device (available from Arcturus, Mountain View, Calif.), laser "shots" are used to fuse a plastic film to the desired cells. This allows the separation of the selected cells from the unselected cells thus resulting in the pure population. Individual shots must be fired over and over again to pick up material. Standard LCM requires a large amount of user input and the process can be extremely tedious.
Often these pure cell populations produced by LCM are used as input material for protein identification and purification schemes in order to isolate and attain samples of the proteins being expressed by the pure cell populations.
By coupling the imaging and identification capabilities of the method 100 to a LCM device, the entire process can be automated. Images of samples containing mixed cell populations are segmented into spectrally homogeneous populations based on the reference images. The resultant segmented images can then be used to guide appropriate stage movements and laser actions in order to accomplish the microdissection procedure. The resulting pure cell populations can be fed directly into protein purfication or analysis systems such as protein mass spcetroscopy.
A number of other areas are suitable for automation. For example, the method can automate the quantification and extent of conditions or disease in tissue that has already been diagnosed. In conditions such as fibrosis, the method can survey a tissue sample identifying the cells or regions that are affected. After identifying the regions, the method can automatically compute the extent of fibrosis in the tissue sample. Similar qunatitative evaluation can be automated. For example, in the evaluation of chronic organ rejection the inflammatory process can be detected and its intensity graded.
We now describe specific implementations for the spectral illuminator. FIG. 3 shows a first implementation of a spectral illuminator 1. LED lamps 10 a-j produce light 110 of various wavelengths that span the spectral range of interest (normally the photopic visible). Lenses 11 a-j collimate the light from each LED to form light 111 and bandpass filters 12 a-j provide spectral filtration of the various bands so that the light 112 has in each band a very well-defined spectral range with much greater specificity than that defined by the LEDs themselves. Objective 13 re-images the collimated light 114 from the various LED channels onto the tip 14 a of optical fiber 14. When light emerges from this fiber end 14 b, it is imaged by lens system 15 and directs light 115 towards the sample without spectro-spatial artifacts.
The illuminator in the FIG. 3 has ten spectral channels, but this is meant to be illustrative rather than restrictive, and systems may be built with any number of channels utilizing the same principles. The beam from each LED is collimated to produce a pupil plane at infinity, oriented parallel to a common optical axis z. The various LEDs and collimators are tiled at the entrance aperture of a telescope, which images all rays of a shared propagation direction to a common point. An optical fiber is placed at that point. The telescope causes the spatially distinct but parallel rays from the various LEDs to pass through a single point with various angles. The optical fiber randomizes the angular distribution, provided that its length is long enough that all rays undergo multiple reflections within it.
While a range of configurations are possible, most commonly the spectral channels will be in the range of 3 nm-20 nm per band, and there will be from 8 to 80 spectral bands overall. LEDs are not presently available with spectral widths narrower than about 35 nm, so it is normally necessary to use interference filters to further define the passband from each given LED. For example, in FIG. 3, there are 10 LEDs and interference filters with center wavelengths spanning the range 430 nm-655 nm by 25 nm intervals. Since the light is collimated at the point it passes through the bandpass filters, the spectral properties of those elements are not compromised.
The combination of the LED collimator lens and the telescope comprise a magnifier with gain F2/F1, where F2 is the focal length of the telescope and F1 is the focal length of the collimator lens. The design of this assembly presents optical engineering problems as will be known to one skilled in the art of optical design. Primary goals are to produce a high efficiency in coupling light from the LED into the fiber, and to ensure the design is manufacturable without requiring unduly tight dimensional tolerances.
Use of a large diameter optical fiber is often favored since this enables capturing more light. Similarly, use of optical fiber with relatively high numerical aperture (NA) is favored in many designs, since the fiber NA limits the effective NA of the telescope. Higher NA means a shorter telescope may be employed, to produce a more compact assembly.
Choice of particular LEDs is dictated by the need for spectral coverage and high brightness. The latter is important since the overall brightness of the illuminator is proportional to the spectral radiance of the LED. It is possible to use bare LEDs of the type that emit vertically (normal to the chip surface), as well as to use packaged LEDs. Choice of one over the other will be dictated by the properties of available LED sources, as well as cost, ease of assembly, and other practical factors. Suitable LEDs include high-brightness LEDs from Stanley Electric (London, Ohio), Cree Research (Durham, N.C.), Nichia (Tokushima, Japan), and KingBright (City of Industry, Calif.). These are sometimes hyperbolically described as "ultrabright", "superbright", and the like. Cree provides LEDs at 430, 450, 470, 490, 505, and 525 nm, which between them cover the range 420-540 nm. Many manufacturers provide LEDs covering the range 525-680 nm and the near-IR range, as is well-known in the art, and one source of such LEDs is MarkTech Optoelectronics (Menands N.Y.). When packaged LEDs can be used, the rays leaving the LED are refracted somewhat by the package, and it may be helpful in some cases to polish the LED front to be flat instead. The collimating lenses 11 a-j are designed accordingly with the goal of maximizing the energy that is coupled into the fiber.
In a related embodiment, the lenses 11 consist of 19 individual lenses, each with 25.4 mm focal length and diameter of 12.7 mm. They are tiled in a hexagonal close-packed arrangement within an overall diameter of 63.5 mm. Nineteen LEDs having wavelengths from 420-690 nm arranged by 15 nm intervals are situated behind the lenses, and nineteen interference filters having corresponding passbands are placed in front of the lenses. Mechanical mounting of the lenses and filters is accomplished using the space between the circles, so maximum packing density is achieved. The telescope consists of a lens with 65 mm diameter and 75 mm focal length, for a working N.A. of 0.397. It couples the collimated light from these LEDs into a 3M multimode fiber with 1 mm core and an NA of 0.39 (ThorLabs, Newton N.J., part FT-1.0-UMT). The fiber length is 2 meters, and at the distal end an achromatic lens of 25.4 mm focal length and 22 mm diameter images the fiber tip to infinity. The output beam may be directly coupled into microscopes such as the Zeiss AxioPlan (available from Carl Zeiss, Thornwood, N.Y.).
In many cases, it is preferable to use a lens array rather than individual lenses for each LED. The lenses can be plastic or glass, as long as the optical quality does not degrade the efficiency of optical coupling into the fiber. Mirrors may be used instead of lenses at each and every instance, and the choice of one over the other will be dictated by cost, optical performance, and other engineering design factors.
It is not essential in the design of FIG. 3 that all LEDs have the same size lens. Varying the numerical aperture (N.A.) of the LED collimation lens provides a means for adjusting the collection efficiency, and hence for tailoring the overall output of the system. Since different LEDs have different spectral radiance properties, this can be desirable. Adjustment of LED drive current is another means for balancing or tailoring the relative output powers of the various channels when each is set to its full-scale intensity setting.
Referring to FIG. 4, a spectral illuminator 1 b is shown. LED lamps 10 a-j produce light 110 of various wavelengths that span the spectral range of interest (normally the photopic visible). Lenses 11 a-j collimate the light from each LED to form light 111. The light passes through a linear-variable filter. These are available from OCLI (Santa Rosa, Calif.) or Reynard Coatings (Marina Del Rey, Calif.). The light 112 has in each band a very well-defined spectral range with much greater specificity than that defined by the LEDs themselves. Objective 13 re-images the collimated light 114 from the various LED channels onto the tip 14 a of optical fiber 14. When light emerges from this fiber end 14 b, it is imaged by lens system 15 and directs light 115 towards the sample without spectro-spatial artifacts.
In this arrangement, the lenses, while approximately spherical or aspherical in surface figure, may preferably have a rectangular shape with a longer axis transverse to the direction of the LED array. This enables better optical efficiency, since they can collect over a greater area.
It is possible to use other elements in concert with, or instead of, the optical components described above to couple the light from several LEDs into a single beam, and to homogenize the beam. Suitable elements include without limitation liquid-type light guides, or spatial scramblers comprising a tube-like enclosure whose internal surfaces are reflective. The latter can be constructed from a glass cylinder whose inner surface is silvered, or four strip-shaped first-surface mirrors arranged to form a rectangular cylinder, or a similar arrangement. In any case, the length of the tube should be several times the width of the tube, in order to sufficiently perform the function of the optical scrambling.
The use of fiber couplers to merge the various sources also eliminates or simplifies some of the alignment problems and enables a modular design, where each source (LED, collimating optics, and bandpass filter) is coupled into a fiberized connector, into which the fiber coupler inputs are plugged. Referring to FIG. 6, a related embodiment of this invention is shown. It is intended for fluorescence excitation uses. LED lamps 10 a-j produce light 110 of various wavelengths that span the spectral range of interest (normally the photopic visible). Lenses 11 a-j collimate the light from each LED to form light 111 and bandpass filters 12 a-j provide spectral filtration of the various bands so that the light 112 has in each band a very well-defined spectral range with much greater specificity than that defined by the LEDs themselves. The light is then coupled into connectors 18 a-j by coupling optics 17 a-c. A fused fiber coupler 19 from Gould Fiber Optics (Millersville Md.) has input fibers 20 a-j and output fiber tip 21. When light emerges from this fiber end 21, it is imaged by lens system 15 and directs light 115 towards the sample without spectro-spatial artifacts. The number of LED sources is M and the fiber coupler has N inputs. This enables use of N inputs at any one instant, under electronic control, in a few microseconds, with no moving parts. Each source can further be modulated in order to perform time-resolved measurements, as is known in the art.
If M equals N, then all bands are accessible at any time. However, this requires an M-fold fiber optic coupler, which becomes expensive for M greater than 2 or 4. While it is unusual for a given experiment to require more than 4 excitation bands, it is common that a researcher will require a wider range of spectral bands over the course of various experiments. This embodiment provides connectors for each LED source, into which a fiber can be inserted. This enables a user to select which of the M bands are of interest for a given experiment, and to plug the fiber coupler input into the appropriate bands. The arrangement is analogous to a telephone patch-bay, where plugging in to the proper jack may access various sources.
Further, the system exhibits greater optical efficiency compared to an arrangement which places all M sources in the pupil plane of the telescope, as the previous embodiments do, since it better utilizes the numerical aperture of the fiber into which the sources are coupled (or equivalently, of the numerical aperture of the telescope which merges their signals). Significantly, the instrument is instantly reconfigurable in the field by the customer without alignment problems. So it is practical to provide a wide array of source wavelengths, out of which only a few are selected for a given experiment, by means of the fiber-optic patch-bay.
FIG. 6 shows another implementation of a spectral illuminator Id where LED lamps 10 a-j are placed at the focal plane of a spectrometer comprising reflective grating 31, lens system 32, and optional exit slit 33. Light emerging from the LEDs is spectrally selected by the grating according to the grating equation and the position of the LEDs, so that only a precisely regulated band enters multimode optical fiber 14 and passes to the sample through lens system 15.
Another approach which may be used to realize a spectral illuminator is pictured in FIG. 7. A lamp 50 and lens 55 illuminate a slit 56 that is imaged to infinity by lens 57. Collimated rays 58 pass to prism 51 where they are dispersed according to wavelength and re-imaged by lens 59 to produce a spectrum at programmable mask 52. This element has a linear array of stripes arranged along an axis that is essentially parallel to the dispersion axis of the spectrum, where each stripe has an electrically variable transmission. Such an element can be constructed using a liquid crystal modulator, where each stripe is a single pixel with a transmission that is altered by the action of the liquid crystal material. These are commercially available from a number of manufacturers, including Cambridge Research and Instrumentation (Woburn, Mass.). The model SLM-128 amplitude mask with integral polarizers is one example of a suitable programmable mask, but any device that provides electrically variable transmission elements in a linear array is suitable. High peak transmission and a wide dynamic range of adjustment are key parameters for this device. Each stripe of the mask 52 controls the transmission for a selected spectral band, and by adjusting the mask it is possible to transmit all, none, or any precisely selected mixture of wavelengths. The light passing through 52 is re-collimated by lens 151 and passes through a second prism 53 that is oriented with its dispersion opposite to that of prism 51. Light emerges from prism 53 in a collimated state, and is re-imaged by lens 152 onto a linear fiber array 54. The fibers in this array are formed into a circular pattern at the other end, where lens 154 couples light 155 from the fibers towards the sample. Equivalent systems can be constructed using gratings in place of the prism elements 51 and 53. Other embodiments of the spectral illuminator can include using diffraction gratings, electronically tunable filters such as acousto-optic tunable filters (AOTFs) or liquid crystal tunable filters (LCTFs).
The overall scheme of the electronics 40 is illustrated in FIG. 8, which shows a digital control module 42 that communicates with external control means via an interface 45 and generates signals 44 a-44 j by means of circuits 43 a-43 j. Optional digital storage element 41 contains drive information for one or more spectral weighting functions. When the spectral illuminator is used in conjunction with a detector of some kind that has linear response, it is possible to use the detector to calibrate the illuminator. This makes use of the linear detector response to generate an in-situ calibration of the possibly nonlinear illuminator.
Additional embodiments and details of suitable multi-spectral illuminators and multi-spectral illuminator systems are described in commonly owned U.S. utility application Ser. No. 10/163,233 by Paul J. Cronin et al., filed Jun. 4, 2002 and entitled "Multispectral Imaging System," the contents of which are incorporated herein by reference.
The use of medical examples is not meant to suggest that this is the sole or predominant use of the present invention. The method described is also suitable for the use of borescopes for industrial inspection, in reflected-light microscopy for semiconductor and materials measurements, and in any endeavor where spectral signatures enable discrimination between sample regions or quantification of sample properties. One such application is in the field of counterfeit detection, since in general the dyes, paints, and materials used in a counterfeit will not match the genuine article at all wavelengths even if the overall visual appearance is quite similar. A spectral weighting function that maximally illuminates in the spectral regions where the genuine article and the fake are different, will highlight these distinctions and make them readily apparent. The present invention provides a means for at once identifying the weighting function (through taking of a spectral image cube and its subsequent analysis on a computer), and then providing essentially real-time imaging of the objects using that derived spectral weighting function (by having the spectral illuminator provide the weighted illuminants while imaging the sample).
A number of embodiments of the invention have been described. Nevertheless, it will be understood that various modifications may be made without departing from the spirit and scope of the invention. Accordingly, other embodiments are within the scope of the following claims.
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Owner name: CAMBRIDGE RESEARCH AND INSTRUMENTATION, INC., MASS
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Maybelline One by One Volum' Express [DISCONTINUED]
768 reviews
64% would repurchase
Package Quality: 4.0
Price: $$
Package Quality: 4.0
Price: $$
INGREDIENTSAqua/Water, Paraffin, Potassium Cetyl Phosphate, Cera Alba/Beeswax, Cera Carnauba/Carnauba Wax/Acacia Senegal Gum, Glycerin, Cetyl Alcohol, Hydroxyethylcellulose, Phenoxyethanol, Peg/Ppg-17/18 Dimethicone, Steareth-20, Phenethyl Alcohol, Sodium Polymethacrylate, Hydrogenated Jojoba Oil, Hydrogenated Palm Oil. Simethicone, Disodium Edta, Polyquaternium-10, Saccharum, Officinarum Extract/Sugar Cane Extract, Ascorbyl Palmitate, Panthenol, Acrylates Copolymer, Silica, Citrus Medica Limonum Peel Extract/ Lemon Peel Extract, Pyrus Malus Extract, Camellia Sinensis Extract, Camellia Sinensis Leaf Extract [+/- May Contains: CI 77491, CI 77492, CI 77499/Iron Oxides, CI 77007/Ultramarines, CI 77891/Titanium Oxides, Mica, CI 75470/Carmine, CI 77288/Chromium Oxide Greens, CI 77289/Chromium Hydroxide Green, CI 77742/Manganese Violet, CI 77510/ Ferric Ferrocyanide] F.I.L. D43811/1
Age: 18 & Under
Skin: Combination, Fair-Medium, Not Sure
Hair: Brown, Other, Other
Eyes: Brown
This is a great lengthening, seperating, and a little volumizing mascara. It gives you a very natural look, and won't clump when trying to build it up. The only down side to this mascara is that if you curl your lashes before hand, it weighs them down, and even if you tryed curling them afterwards, they would still droop again. I do have very hard to curl lashes, so that might be why. The brush is amazing and the formula is on the wet side, which I don't have a problem with, but with some people that might push them away. Overall, I definitely recommend purchasing this mascara.
Age: 18 & Under
Skin: Combination, Fair, Cool
Hair: Brown, Straight, Other
Eyes: Grey
This is such an amazing mascara, it does everything a proper mascara should, imo.
It separates my lashes so well, lenghtens and volumazes, and makes them look totally great! I have long thick lashes naturally, but this mascara makes them look like I'm actually wearing false ones! You can get a very natural feathered look, as well as big voluminous super long lashes look! Ah I love this one so much! I would suggest it to anybody!
Age: 30-35
Skin: Acne-prone, Fair-Medium, Not Sure
Hair: Red, Other, Other
Eyes: Brown
Bought it based on reviews I saw here. Mascara junkie. Bought this at the same time I bought Benefit They're Real! I would but 3 of these for the price of the Benefit mascara. Love this formula. Wetter...makes it able to work with. NO clumps. While swiping upwards I hold for about 15-20 seconds. LOVES it!
Age: 19-24
Skin: Normal, Fair, Not Sure
Hair: Brunette, Wavy, Medium
Eyes: Hazel
This did nothing for my lashes.
The formula was too wet and when I applied it to my lashes it felt heavy. Worst mascara ever.
Age: 36-43
Skin: Combination, Fair, Neutral
Hair: Brown, Straight, Fine
Eyes: Blue
PROS: Kinda meh for me, but I have very sparse and thin lashes. Would probably be good for someone who has good lashes to begin with.
CONS: I couldn't layer it enough to give me good results.
ALTERNATES: Milani Runway Lashes is gorgeous for length/volume. I cover it with Clarins Double Fix to keep it from smudging. The Waterproof version of the Rocket by Maybelline is wonderful for volume and lots of length. For lots of volume and good length, I like Covergirl Remarkable Washable Mascara, which stays on very well for nonwaterproof.
APPLICATION TIPS: I am a recent convert to using Shiseido's popular curler
Age: 18 & Under
Skin: Combination, Medium, Not Sure
Hair: Black, Other, Other
Eyes: Black
I got this mascara because I thought the brush would separates. It looks like it would separates but instead it would clump your lashes into 3 . Worst mascara ever.
Age: 25-29
Skin: Very Oily, Fair-Medium, Not Sure
Hair: Brunette, Other, Other
Eyes: Hazel
Amazing mascara. My favorite by far. Better than high end ones even my dior one which used to be my favorite untill this one. Makes my lashes sooooo long without clumps! Seriously they've never looked better and i get so many complimants when I use this mascara! I do use it with a dior lash primer though.
Age: 19-24
Skin: Combination, Fair-Medium, Not Sure
Hair: Brunette, Other, Other
Eyes: Blue
I absolutely love this mascara. I have it in black, it works great for me. Although I don't really like the scent of it I love the brush. These types of brushes definitely work best for me and catch every lash. It doesn't give me clumpy spider lashes, they're very defined and spread out. I like it so much I've already bought another and I'm on my second tube. Great product. I sometimes wear it over my Physician's Formula Fakeout for a little more pop.
Age: 18 & Under
Skin: Combination, Tan, Not Sure
Hair: Black, Other, Other
Eyes: Black
I don't know why i kept re-purchasing this mascara, i had been using it over a year now. It has very little lengthening power although the volume is amazing and does a terrific job at separation of the lashes, it kind of reminds me of the newer volume express mascara the rocket volume, it did the same effect which I'm not surprize because there is a reason why the line is called "volume express" it's for volume. I have always notice the mascaras from the line never really lengthen my lashes just volume which is such a bummer. i will no longer re-purchase this product.
Age: 19-24
Skin: Normal, Fair, Cool
Hair: Brown, Wavy, Medium
Eyes: Green
This mascara is hands down my favorite Maybelline mascara. My all time favorite is Max Factor's 2000 calorie in Rich Black but sadly this line has apparently been discontinued. So on the hunt for a new drugstore mascara I eventually stumbled upon Volume Express One by One. I believe I purchased it at Walmart for around $6.
This mascara does not clump, AT ALL. It seperates my lashes perfectly. I could probably apply 10 coats and still be clump free. It does take about 3 coats to achieve the volume and fullness that I like.
I have 4 empty tubes in a drawer that I haven't tossed out yet. Will be repurchasing again.
My Max Factor 2000 calorie mascara is still greatly missed though...that stuff was by far the best.
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subset: commoncrawl


text:
The Merritt Island boys soccer team defeated Sunlake 1-0 in Thursday's Class 3A state semifinals.
- Filed Under
Class 3A state semifinal
Merritt Island 1, Sunlake 0 ...
Merritt Island 1 0 - 1
Class 3A state semifinal
Merritt Island 1, Sunlake 0 ...
Merritt Island 1 0 - 1
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Lawmakers Say U.S. Faces Loss of Credibility; No Weapons of Mass Destruction Found So Far in Iraq
Byline: Audrey Hudson, THE WASHINGTON TIMES
Key Republican and Democratic lawmakers yesterday said the U.S. faces credibility problems if weapons of mass destruction are not found in Iraq, but they are optimistic such weapons will be uncovered by a new survey team.
Sen. Joseph R. Biden Jr., Delaware Democrat and ranking member of the Senate Foreign Relations Committee, said Mr. Bush unnecessarily "hyped" the extent of weapons in Iraq's arsenal.
"I do think that we hyped nuclear, we hyped al Qaeda, we hyped the ability to disperse and use these weapons," Mr. Biden said on NBC's "Meet the Press."
Such hype is customary for all presidents "when they want to accomplish a goal that they are trying to get broad national support for," but it was not needed in this case, Mr. Biden said.
"We didn't need all that to go into Iraq. There was sufficient evidence to go into Iraq. They violated the United Nations' security agreements. They, in fact, violated essentially a peace treaty," Mr. Biden said.
Mr. Biden called the hype a "serious mistake" but said, "I do think we'll find weapons of mass destruction."
Sen. Pat Roberts, Kansas Republican and chairman of the Senate Select Committee on Intelligence, said weapons must be found to avoid "a real credibility problem."
"We are now putting in 1,400 people with an Iraq survey group, as opposed to the tactical people that are there now, and it's my view that we will find out what happened to the weapons of mass destruction," Mr. Roberts said.
Sen. Robert C. Byrd, West Virginia Democrat, last week accused the Bush administration of deliberately misleading the American public into war with Iraq because weapons have not been found.
"What has become painfully clear in the aftermath of war is that Iraq was no immediate threat to the U.S.," said Mr. Byrd, who also supported the resolution.
Rep. Jane Harman, California Democrat and ranking member on the House Intelligence Committee, said on CBS she believed the prewar information and does not believe intelligence was wrong.
However, she said, "the world is owed an accounting."
"And frankly, I'd like to know other things about what did we know about the Iraqi will to fight, which seemed stronger than predicted; what did we know about the possibility of riots and looting, which emerged; what did we know about the intentions of the leadership of Iraq?" Mrs. Harman said.
Since the war ended six weeks ago, two mobile labs that could have manufactured biological weapons have been found, but there has been no sign of the weapons or ousted leader Saddam Hussein. …


text:
Farm Engineer Was an Expert in His Field; George Elkington May Have Been an Innovator in the Industrial World of Birmingham but His Lesser Known Grandfather Made His Own Mark on History Too, Writes Chris Upton
Byline: Chris Upton
George Elkington is one of the more celebrated of Birmingham's industrial innovators, famous enough to warrant a blue plaque on the front of his old factory in Newhall Street (later the home of the Science Museum). Elkington's pioneering use of electro-plating, and his development of celluloid, placed him in the forefront of technological advances in the mid-19th Century.
What is less well known is that originality was deep in the Elkington genes. Two generations earlier, and in very different circumstances, George's grandfather was making his own mark in history. There could not be a better link between England's two revolutions - one agricultural and one industrial - than the tale of two Elkingtons.
Joseph Elkington lies buried in the churchyard at Madeley in Staffordshire, where he owned (or rented) some 500 acres of land, living at Hey House, until his death in October 1806. But it was in Warwickshire, the county of his birth, that Joseph started his own little revolution.
Joseph Elkington's home territory was at Princethorpe on the Fosse Way, a few miles south of Leamington Spa. It was at the nearby church of Stretton on Dunsmore that he was baptised in 1740, and in its churchyard he is commemorated by a monument, restored by the Warwickshire Agricultural Society back in the 1960s. "Pioneer of land drainage" the memorial calls him.
Drainage might not look like a crucial element in good husbandry, but ask any farmer - from the 18th Century onwards, if you can find one - and they'll tell you that having just the right amount of water on one's land is vital.
The trouble is that our water sources are not evenly spread or conveniently located. Under the surface of the soil hidden springs and watercourses make their way, bubbling up in some places, diving deep in others. A clay soil only serves to trap the water in great underground reservoirs, spilling out on to the surface. The presence of an underground watercourse can easily turn a field into a boggy morass, unsuitable either for crops or for grazing animals.
Joseph Elkington had a boggy field just like this at Princethorpe, and set about working out how to deal with it. Like all great discoveries, it was really quite simple.
Our account of what Elkington did comes from a report commissioned by Parliament in 1797, some 30 years after the event. To begin with, Elkington dug a trench, some four or five feet deep, alongside the bog, in an effort to detect the source of the water. But the source was clearly much further down than this. Elkington's contemporaries may have done much the same as this, cursed their bad luck and moved on to another field.
Only when Joseph drove a crowbar down into the clay at the bottom of the trench did the water gush forth. And then a drain could be cut to take the water away.
So Elkington's method was to use boreholes to intercept the source of water. …


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Pressure-packed regular season one of the benefits of BCS
My hair is graying, and I've got a disk in my back that looks at my golf bag and laughs. I've got a 10th-grader who's learning how to drive and a fifth-grader who wants to know when she's getting a cell phone. You want to talk aging?All of those symptoms are fixable (and the answer on the cell phone is still no). But when I sit and think about why I not only can live with the BCS but that I want to, I realize that the reason is simple maturity. See the value in ...
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The BCS at 10
• Maisel: Paradox found
• Low: Best BCS bowl games
• Miller: Best BCS bowl performances SportsNation
• Rank 'em: Best BCS bowl games
• Rank 'em: Best BCS bowl performances
• Chat wrap: Chris Low
• Chat wrap Ted Miller
• Maisel: Say yes to the BCS
• Forde: Say no to the BCS
• Miller: Biggest BCS bowl slights
• Rank 'em: Biggest BCS bowl slights
• Chat wrap: Ivan Maisel
• Chat wrap: Pat Forde
• Schlabach: Ranking the BCS champions
• Schlabach: Defining coaches of the BCS era
• Rittenberg: Memorable BCS bowl coaching moves
• Rank 'em: Which BCS champion is the best?
• Rank 'em: Which coach defines the era?
• Chat wrap: Adam Rittenberg
• Chat wrap: Mark Schlabach
Conference Breakdowns
Dinich: ACC has been a dud in BCS era
Big 12
Griffin: Big 12 becomes superpower in BCS era
Big East
Schlabach: Big East thriving in BCS
Big Ten
Rittenberg: BCS system has been good to Big Ten
Miller: USC dominates Pac-10 in BCS era
Low: SEC successfully navigates BCS system
Rittenberg: No complaints from South Bend
• Chat wrap: Tim Griffin
• Schlabach: The future is now
• Griffin: Defining players of the BCS decade
• Dinich: Memorable BCS bowl moments
• Rank 'em: Defining players of the BCS era?
• Rank 'em: Memorable BCS bowl moment?
• Chat: Heather Dinich, 2 ET


text:
Lights Up Theatre Season
Theatre Season
2016 Theatre Season Launch2 Dec 2015 - 6:00pm
Theatre Season
Christmas Gift Vouchers25 Dec 2015 - 1:00pm
Like the filament of a light globe glowing intensely, we know our Lights Up 2015 season shines the light on a year of rich and varied performances. The 2015 selection has been carefully chosen to bring you extraordinary entertainment and wonderful performances.
In 2015 we will explore and celebrate our connection to the land and history through performances that resonate with us as Australians. Matthew Ryan's new play Kelly dramatises the last night of the iconic bushranger in a gripping and tautly written piece. We are thrilled to include celebrated local musician Shane Howard in an intimate concert performance of his new album, Deeper South. Australia Day written and directed by comic genius, Jonathan Biggins, pokes fun at Australian traditions, local politics and sacred cows.
Miriam Margolyes, international star of stage and screen will grace our stage in her own flamboyant, theatrical celebration. Michael Veitch brings to life the heroics of the pilots of World War II in a moving and very personal account. Amity Dry (best known from "The Block") delights in her own musical creation Mother, Wife and the Complicated Life.
We continue the family fun with a special pyjama performance of The 26 Storey Tree House and the return of 360 Allstars the exciting, amped up Urban Circus after rave reviews and sell out seasons in Europe and America.
We also welcome back some of our favourite performing arts companies and artists - Australian Chamber Orchestra 2, Bell Shakespeare as well as the delightful Bethany Simons and the irrepressibly comic Damian Callinan.
The 2015 selection has been carefully picked especially to have something for everyone and the chance to enjoy it right on your doorstep. Become a subscriber in 2015 by choosing four or more shows from the season and enjoy the rewards – priority booking, cheaper tickets, VIP functions and deferred payment.
The Lights Up 2015 season glows with fun, vibrancy and entertainment.
We're excited, and hope you will be part of it!


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Part self-help, part autobiography, this book is all about the power of positive thinking. It's also the account of how one suddenly-single grandmother with a great sense of humor came to find herself through caring for her grandson while his parents were deployed overseas by the U.S. Military. A heartwarming tale of love, devotion and inspiration.
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Other writing by reportingfordoodie Reporting for Doodie: Toy Story
Life, Death, Book, Fiction, Non-fiction, Love, Books, Spirituality, Story, Religion, Self-help, Help, Universe, True, Evolution, Relationships, Creation, Nonfiction, World, Living, Romance


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Analysts cite abundant fuel supplies, lower demand
Higher supply, declining demand, and a little tax break from the state have pushed the average price for a regular gallon of gasoline in San Diego County below $4 for the first time since March.
On Monday, the price for a gallon of regular unleaded here cost $3.97, down four cents from a week ago, but a larger drop from the average $4.14 per gallon price a month ago, the Auto Club of Southern California reports.
The price for a gallon of gas fell below the $4 threshold Friday. The highest price for gasoline this year has been $4.29 on April 29.
Charles Langley, a San Diego-based gasoline analyst, said an oversupply of gasoline is causing the drop.
"There's more surplus being made available now to unbranded, independent stations. They're lowering the price and it's making it difficult for the major brands to compete without lowering their price," he said. "As soon as that situation changes, the price will go up."
Langley said a huge spike in oil prices would change the situation, but that oil has been on the decline. For instance, a barrel of the benchmark West Texas Intermediate oil dropped to $101.71 on Monday. Langley said he was fascinated that the shooting down of Malaysia Airlines Flight 17 over Ukraine, and the unrest in Israel and Gaza have not caused oil prices to jump.
"All that stuff was really a good excuse for the market to rally and to drive oil prices up high, and that did not happen at all," he said. "It suggests to me that oil supplies are relatively abundant, especially here in the U.S."
Gregg Laskoski, senior petroleum analyst at Gasbuddy.com, which has its own data that follows a similar pattern, said basic fundamentals are contributing to the drop in prices, which the website reports to be $3.95 per gallon.
"We have a very healthy supply, refinery operations are doing very well right now, and at the same time, even though we're in the summer driving season, consumer demand has not been remarkable," he said.
Laskoski said Americans are driving less as they age, and also newer cars are more efficient, keeping demand down. He said the state in September will switch to its winter blend of gasoline, which should push prices lower, wherever they may be at that time.
Also helping push the price just below $4 is the state Board of Equalization lowering the excise tax on gasoline by 3.5 cents per gallon on July 1, the start of the fiscal year.
Fuel prices were last this low on March 15, when a regular gallon of gas cost $3.99.
A year ago, San Diegans were paying $4.05 per gallon.
In Southern California, only Los Angeles remains above $4 per gallon on average. Nationwide, the average gallon of gasoline costs $3.52, down from $3.63 a year ago.


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Despite Reaper beating 90210 in the ratings, it looks like The CW will be losing its funniest supernatural dramedy. No offense, Supernatural, you just don't have Sock or Ray Wise as the Devil.
According to Zap2It, Michele Fazekas and Tara Butters - the creators of the hilarious show - have signed a seven-figure two-year-deal with 20th Century Fox that asks them to leave the series behind.
So without its creators and producers, how much faith do you have in Reaper's return? It's currently in its second season and has yet to be renewed.