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US5073340A - Depositing a binder on a solid support - Google Patents
Depositing a binder on a solid support Download PDF
Gloria J. Covington
Timothy G. Bloomster
Becton Dickinson and Co
1990-07-06 Application filed by Becton Dickinson and Co filed Critical Becton Dickinson and Co
1991-08-26 Priority claimed from US07/749,734 external-priority patent/US5204061A/en
G01—MEASURING; TESTING
G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
G01N33/54366—Apparatus specially adapted for solid-phase testing
A test device for use in determining analyte wherein binder is supported on a solid support in admixture with a marker whereby the presence and location of binder on the support can be determined prior to the assay. A control for the assay may also be applied to the support in admixture with a label.
This application is a continuation of application Ser. No. 106,075, filed Oct. 8, 1987, now abandoned.
This invention relates to a test device useful in an assay for an analyte and to the use thereof in such assays. More particularly, the present invention relates to providing a binder for an analyte on a solid support for use in a solid phase assay for an analyte.
Assays for determining analytes by a solid phase technique involve contact between a sample suspected of containing the analyte and a test device having a binder specific for the analyte supported on a solid support. The amount of analyte which becomes bound to the binder on the solid support is then detected with a tracer as a measure (quantitative or qualitative) of analyte in the sample.
In some solid phase assays, for example, as described in U.S. Pat. No. 4,632,901, the binder for the analyte is supported on a limited portion of the solid support in order to provide a test area and an area surrounding the test area which is free of binder. The area free of binder may be employed as a background area to aid in determining of analyte by the use of a suitable tracer. When the test device includes multiple layers, they need to be assembled with the test area oriented correctly relative to the other components of the device. Obtaining proper registry is difficult when the binder is not visible.
Use of a marker in admixture with a binder allows construction of test devices where a test area of a solid support having binder on it is distinguishable from a background area surrounding the test area. The test device of the present invention is for use in an assay for an analyte. It has a solid support having a test area and a background area. A mixture of binder for the analyte and a marker is supported on the test area. The test kit of the present invention includes the test device and a tracer having a tag which is distinguishable from the marker.
The assay of the present invention employs a test device having a solid support with a test area and a background area. The test device has a mixture of a binder for an analyte and a marker supported on the test area. The test device is contacted with a fluid sample and a tracer. The tracer has a tag which is distinguishable from the marker.
In the manufacturing method of the present invention a solid support which includes a test area and a background area is provided. A mixture of binder and a marker is applied to the test area so that it is supported there. In this manner the marker is useful to determine that binder has in fact been applied to the test areas. The method facilitates assembly of the solid support with other components of the test device without compromising performance of the binder under assay conditions.
The marker which is employed in admixture with the binder is one which does not interfere with the binding properties of the binder, is capable of being supported on the solid support in a manner similar to the binder, and does not interfere with the ability to detect tracer in the test area.
In accordance with a preferred embodiment, the binder is supported on the solid support by adsorption; accordingly, the marker which is employed in admixture with the binder is one which is also capable of being adsorbed by the solid support.
The preferred marker is a material which under appropriate conditions is detectable with the naked eye. The marker may be a colored material which absorbs light of a characteristic band within the visible spectrum. Alternatively, and preferably, the marker is a fluorescent material which upon excitation emits a characteristic fluorescent signal in the visible region of the spectrum. Colored and fluorescent materials are referred to hereafter as "chromogens." With these chromogens, application of the mixture of binder and marker to the test area of the solid support may be easily determined by the presence of color or upon excitation by the presence of a fluorescent signal in the test area.
The marker which is preferably a chromogen, is applied to the solid support in admixture with the binder in an amount which does not adversely affect the binding properties of the binder, and which is sufficient to permit detection of the marker in the test area. The marker is preferably water soluble because in most cases a binder is applied to a solid support in a water based solution.
Thus, by proceeding in accordance with the manufacturing method of the present invention, wherein a binder is applied to the test area of a solid support in admixture with a marker, a test area of the support having binder securely attached it can be distinguished from a background area. In this manner, if the solid support is to be located in a container which includes an inlet port for applying a sample and other materials to the test device, the test area in which the binder is supported can be properly located with respect to the sample inlet port.
In some cases, application of a control (for example an analyte positive control) to a solid support is desirable. In accordance with another aspect of the present invention, the control (in particular a solution containing a known amount of the analyte) is applied to a control area of the solid support in admixture with a label, which is distinguishable from the marker whereby the control area may be determined by detecting the label. In accordance with this aspect of the present invention, the control area may be completely or partially overlapping with the test area or it may be a completely separate area of the support. In most cases, the control is a positive control; however, a negative control may also be applied.
Thus, in accordance with the present invention, the label which is applied to the solid support in admixture with a control is distinguishable from both the tag employed in the tracer used in the assay and from the marker applied in admixture with the binder. The label which is applied in admixture with the control has the characteristics described above with respect to the marker; i.e., the material is one which does not interfere with the binding capabilities of the control, which is capable of being supported by the solid support in a manner similar to the control, which does not interfere with the interactions occurring under assay conditions, and which is distinguishable from the tracer under assay conditions. The label employed in admixture with the control is also preferably a chromogen and most preferably a fluorescent material.
The present invention is useful in any assay where a binder is supported on a solid support. The choice of binder is dependent on the assay format and protocol. The selection of a suitable binder is deemed to be within the scope of those skilled in the art.
The solid support may take a wide variety of forms such as, for example, a sheet or membrane, a test strip, a dip stick, a card, or the like. The selection of a suitable form is deemed to be within the scope of those skilled in the art. Similarly, the solid support may be made of a wider variety of materials. Preferably the support is porous.
FIG. 1 is a top view of a test device which incorporates the present invention;
FIG. 2 is a side elevational view of the test device of FIG. 1;
FIG. 3 is a section taken along line 3--3 of FIG. 1; and
FIGS. 4A and 4B are simplified schematic representations of alternative versions of first layer 11 which includes a binder and control.
In accordance with the present invention, a binder which is to be applied to a test area of a solid support, preferably as an aqueous solution or suspension is mixed with a marker, which is preferably a chromogen, and in particular a fluorescent material. The mixture of binder and chromogen is then applied to a defined test area of the solid support, such as, for example, a spot, square, circle, triangle, or any other shape desired, in a manner to support both the binder and the detectable label in the test area. While those skilled in the art are familiar with a variety of coupling techniques to securely attach a binder to a solid support, adsorption is preferred. The procedure for adsorbing the binder can be any of those commonly available for pattern coating of materials including gravure printing, silk screening and other convenional printing methods.
The preferred materials for use as a marker are fluorescent materials which do not adversely affect the binding ability of the binder, and which are capable of being adsorbed by the support. As representative examples of suitable fluorescent materials, there may be mentioned Acridine Orange, Pyronin Y, Texas Red, Rhodamine 6G, with such materials being employed in amounts which do not adversely affect the binding ability of the binder, which are detectable when subjected to ultraviolet light, and which are not detectable with the naked eye. If the material used as a marker includes groups which may be reactive with the binder or control, such groups may be blocked prior to admixing the material, for example they may be placed in a buffer including a source of amino groups (e.g. glycine) that react with reactive sites on the marker and thereby prevent further reactivity which could influence the biological activity of the binder or provide sites for non specific binding under assay conditions.
After the binder has been supported on the solid support, the test area may be distinguished from the background area by detecting the marker. Thus, for example, in the case where the label is a fluorescent material, the support may be exposed to excitation energy of a suitable wave length (e.g. ultraviolet light from conventional U.V. lamps). The marker will emit a characteristic fluorescent signal which is preferably detectable with the naked eye.
In an embodiment employing a control, a solution or suspension of the control and a label which is distinguishable from the marker employed is prepared, with the label preferably being a chromogen, and most preferably a fluorescent material. The solution or suspension of control and label is applied to the solid support in its designated control area, a portion of which or none of which may overlap with the test area. The fact that the control has been applied to the solid support, and the area to which it has been applied, may be determined by detecting the label, for example, in the case of a fluorescent material by exposing the material to excitation energy of a suitable wavelength.
Thus, the binder may be applied to the solid support in admixture with a fluorescent material having a characteristic emission band within the visible spectrum and observable as a first color (such as yellow), and the control may be applied in admixture with a fluorescent material having a characteristic emission band within the visible spectrum and observable as a second color (such as red), which is distinguishable from the first color. After application of both the control and the binder, the solid support may be exposed to excitation energy of suitable wavelengths to detect the presence of both colors to thereby determine the presence, in their respective areas, of both the binder and the control. Preferably the excitation wavelengths for the marker and the label are in the ultraviolet range of the sprectrum and in the absence of U.V. irradiation, they are colorless or are colored with a relatively weak intensity.
The analyte may be determined (qualitatively or quantitatively) by applying a sample suspected of containing the analyte and tracer to the test area. The sample may be applied to an area smaller, equal to or greater than the test area in which the binder is supported. Depending on the chemistry of the assay, reactions take place and presence or absence of a detectable signal from the tracer is indicative of the presence of analyte.
While the chemistry of the assay is independent of the present invention, the preferred assay chemistry uses interactions among specific binding pairs to determine the presence of analyte. The preferred type of specific binding pairs are antigen/antibody pairs.
In the preferred assay chemistry, the tracer is one member of a specific binding pair having a tag coupled to it. For example, if the assay is a competitive format, the specific binding portion of the tracer would be one which is bound by the binder supported on the solid support. If the assay is a sandwich format, then the specific binding portion of the tracer would be bound by the analyte.
Such assay formats and amplification procedures are generally known in the art, and a further description is not required for a complete understanding of the present invention.
The tag portion of the tracer may be any one of a wide variety of materials, including, for example, enzymes, and chromogens. If the tag is detectable by color (as in colormetric enzyme tags and direct detection with a colored material) the color should be observable in the presence of the marker and label. Presently preferred are colored particulate labels, such as liposomes including a chromogen. The selection of a suitable tag is deemed to be within the scope of those skilled in the art.
In the assay technique, the tracer is applied to at least the test area of the solid support, and in the case where a control is used, the tracer is also applied to at least the control area. The tracer may be applied to an area greater than the test area and the control area.
In cases where a positive control is employed, the specific binding portion of the tracer is bound by the positive control whereby the tag of the tracer should be detected in the control area. In addition, if analyte is present in the sample, the tag of the tracer should be detected in the test area.
In accordance with a particularly preferred embodiment of the present invention, the binder in admixture with the marker is supported on a test area located at the surface of a solid support in a concentration whereby a tracer which includes a visible particulate label as its tag portion, under assay conditions, is visible on the support, without further treatment.
The binder supported on the test area is preferably present in at least one microgram per cm2, most generally at least 10 micrograms per cm2, and preferably 40 micrograms per cm2. The residual binding capacity of the test area and the background area may be saturated or blocked by treatment of the solid support with one or more types of proteins which do not specifically bind materials to be employed in the assay. Thus, for example, residual binding capacity may be blocked by use of bovine serum albumin. A wetting agent also may be applied to the test area.
In some cases, in applying the binder (in particular an antibody) to the test area, a polyhydroxy compound (e.g., glycerol, erythritol, and sorbitol), or a sugar (e.g., glucose and sucrose) is included in the antibody solution to prevent non-specific binding (false positives) during the assay.
The solid support which is used is one which has a surface area (area/unit weight of material) such that the binder can be supported on the support in a concentration (weight/unit area) such that the tracer is visible under the assay conditions. The term "visible" as used herein means that the material can be detected with the naked eye without the use of instrumentation; although instrumentation may be used to detect the intensity of the absorbence or fluorescence of the material.
The test area is preferably formed from a cellulose ester with nitrocellulose giving exceptionally good results. The term "nitrocellulose" refers to nitric acid esters of cellulose, which may be nitrocellulose alone, or a mixed ester of nitric acid and other acids, and in particular, aliphatic carboxylic acids having from one to seven carbon atoms, with acetic acid being preferred. Sheets which are formed from cellulose esterified with nitric acid alone, or a mixture of nitric acid and another acid such as acetic acid, are often referred to as nitrocellulose paper.
Although nitrocellulose is a preferred material for the test area (and the entire solid support), other materials having a surface area sufficient for supporting the binder in a concentration as described above may also be employed.
As indicated above, in producing the preferred tracer one member of a specific binding pair is labeled with a particulate label, which is visible. A preferred particulate label is a sac, which includes a color substance whereby the tracer, when used in the assay, is visible without destruction of the sac to release the colored substance.
The sac which is used to label the specific binding portion of the tracer may be any one of a wide variety of sacs, including but not limited to liposomes (single walled or multilamellar) or polymer microcapsules (for example, those made by coascervation, or interfacial polymerization).
Polymer microcapsules also may be produced by procedures known in the art except that the solution in which the microcapsules are formed also includes the tag whereby the interior of the polymer microcapsule includes the tag. The preparation of such microcapsules is disclosed for example in Microencapsulation Processes and Applications, edited by Jan E. Vandegger (Plenum Press, 1974). polymer nucleus coated with such a dye or pigment. Such labels are described in more detail in U.S. Pat. No. 4,373,932, which is hereby incorporated by reference. The tracers produced in accordance with that patent may also be employed as tracers in the present invention.
The visible particulate label may be visible polymer particles, such as colored polystryrene particles, preferably of spherical shape.
Representative examples of other suitable particulate labels include ferritin, phycoerythrins or other phycobili-proteins; precipitated or insoluble metals or alloys; fungal, algal, or bacterial pigments or derivatives such as bacterial chlorophylls; plant materials or derivatives, and the like.
The specific binding portion of the tracer may be labeled with the particulate label so as to produce a tracer for use in the invention by procedures generally known in the art, with the procedure which is used being dependent upon the choice of specific binding species and the particulate label. Such techniques include covalent coupling, derivatization or activation, and the like. In producing a tracer wherein the binder is labeled with a sac, the sac may be produced from a component which has been derivatized with a specific binding species, whereby the sac, when produced, is coupled to the specific binding portion.
Thus, the preferred tracer is comprised of a specific binding portion and a particulate label (solid or solid-like, as opposed to non-solid
As known in the art, liposomes can be prepared from a wide variety of lipids, including phospholipids, glycolipids, steroids, relatively long chain alkyl esters; e.g., alkyl phosphates, fatty acid esters; e.g., leitchin, fatty amines and the like. A mixture of fatty materials may be employed, such as a combination of neutral steroid, a charged amphiphile and a phospholipid. As illustrative examples of phospholipids, there may be mentioned lecithin, sphingomyelin, dipalmitoyl, and the like. As representative steroids, there may be mentioned cholesterol, cholestanol, lanesterol, and the like. As representative examples of charged amphiphilic compounds, which generally contain from 12 to 30 carbon atoms, there may be mentioned mono- or dialkyl phosphate ester or an alkylamine; e.g., dicetyl phosphate, stearyl amine, hexadecyl amine, dilauryl phosphate, and the like.
The liposome sacs are prepared in an aqueous solution including the tag whereby the sacs will include the tag in their interior. The liposome sacs may be prepared by vigorous agitation in the solution, followed by removal of tag from the exterior of the sac.
Further details with respect to the preparation of liposomes are set forth in U.S. Pat. No. 4,342,826 and PCT International Publication No. WO80/01515, both of which are hereby incorporated by reference.
The tracer may also be produced by labeling the specific binding species with an aqueous dispersion of a hydrophobic dye or pigment, or a labels, such as radioisotopes, enzymes and various fluorescent materials), and the particulate label provides a tag which is visible under the assay conditions so that the presence or amount of analyte may be determined without further treatment and without the use of instrumentation; e.g., by the use of a liposome containing a colored material as the particulate label.
In accordance with a preferred embodiment, the test device is constructed as described in commonly assigned U.S. Ser. No. 016,846, now abandoned and Ser. No. 106,757, filed Aug. 8, 1987. The preferred device includes a porous solid support for supporting the binder at a test area in a concentration whereby the tracer used in the assay, when bound to the test area, under assay conditions is visible without further treatment. It also includes a flow control layer, beneath the porous solid support, which is formed of a porous material having a pore size to control the rate of flow of assay reagents through the test area. The test device also preferably includes a porous spacer layer for spacing an absorbent layer, formed of an absorbent material, from the flow control layer.
The absorbent layer has an absorbency sufficient to absorb the reagent liquids applied to the test layer during the assay. In addition, the absorbent materials functions to provide for flow through the test layer.
The entire porous solid support may be formed of the material used in the test area. Alternatively, only the test area may be formed of such a material.
In addition, since the test device is employed in a manner such that the assay reagents flow through its layers, solid support has a pore size which is greater than the size of the particulate label employed in the assay so that portions of the tracer, which do not become bound under assay conditions, flow into the absorbent layer and are not visible at the test area. In general, the solid support should have a pore size which is at least 2 um, and most preferably at least 5 um. In general, the pore size does not exceed 12 um. It is to be understood, however, that although the previously described pore sizes are preferred, other pore sizes may be employed, depending upon the materials used in the assay.
The flow control layer of the test device is formed of a porous material which is employed to control the rate of flow of assay reagents through the test layer and into the absorbent layer. The preferred porous material which is employed in forming the flow control layer has a pore size which is less than the pore size of the material employed for forming the solid support. Thus, in effect, the flow control layer functions to reduce the rate of flow of assay reagents through the more porous test area.
The pore size of the flow control layer, as well as the thickness of the flow control layer, is preferably controlled in a manner such that the flow of assay reagents through the test area provides the requisite sensitivity as well as a rapid and accurate assay.
In accordance with one particularly preferred embodiment, the layer for controlling rate of flow through the test device is dimensioned and sized in a manner such that the flow rate of materials through the test area is in the order of at least 0.5 ml/min, and generally no more than 2 ml/min.
The flow control layer is preferably formed from a non-fibrous material polycarbonate and having pores or channels of a uniform size that provide for unidirectional flow from the test layer to the layer beneath the flow control layer.
Immediately, below the flow control layer of the preferred test device, a spacer layer is provided. It is a porous material which functions as a spacer between the flow control layer, and the absorbent layer. The porous spacer layer has a pore size greater than the pore size of the flow controlling layer so that the spacer layer does not function to restrict flow through the test device.
The preferred test device also includes an absorbent layer which is a porous material having an absorbing or absorbent capacity sufficient to absorb the liquids which flow into the test device during the assay. The absorbent layer also functions to provide a driving force (e.g., a concentration differential) which causes reagents applied to the test area to flow into the absorbent layer.
Thus, in accordance with the preferred embodiment of the present invention, an assay employs a tracer wherein the tag portion of the tracer is a visible particulate sac, and wherein the assay is performed on a test device, which is preferably formed from a plurality of layers of material having different characteristics, as described above, and wherein the assay reagents flow through the test area of the test device.
The materials which are employed in forming the various layers of the test device are selected to have the characteristics described above. In addition, such materials should not produce non-specific binding of analyte or tracer. The materials may inherently have such characteristics, or alternatively, the materials may be treated to prevent nonspecific binding; for example, treatment with an appropriate protein, such as bovine serum albumin. The solid support of the test device is preferably also treated with a wetting agent in order to insure proper flow of the assay reagents through the test layer and into the absorbent layer. Representative examples of wetting agents include sucrose, glycerol, glucose, and sorbitol. The solid support may be simultaneously treated with a protein and wetting agent; e.g., an aqueous solution of bovine serum albumin and sucrose.
In general, the test device is mounted on or in a suitable holder, such as a card or a container. The selection of a suitable holder for the test device is deemed to be within the scope of those skilled in the art.
In addition, the preferred test device is provided with a cover having an aperature, which directs assay reagents to the test area. Thus, for example, the test device may be covered with a card including an aperture which overlies the test area whereby the liquid sample and various assay reagents are applied directly to the test area. Alternatively, the test device may be placed in a container which includes a suitable aperture for directing the sample and assay reagents to the test area. The use of a marker in admixture with the binder identifies the test area whereby it may be properly positioned with respect to the aperture.
Referring now to the drawings, a test device, generally designated as 10, comprised of a solid support, generally designated as 11, which has a test area for supporting a binder. A preferred material is nitrocellulose, which has a pore size in excess of 2 microns, and generally less than 12 microns. Most preferred is a pore size of about 5 microns.
Immediately underneath the layer 11 is a flow control layer 12 which is preferably formed from a unidirectional flow controlling polycarbonate membrane having a pore size of 0.6 microns.
Immediately underneath flow controlling layer 12 a spacer layer 13 is provided. The spacer layer 13 is formed of a porous material, and generally has a pore size greater than the pore size of flow controlling layer 12. The layer 13 may be formed, for example, from a non-woven polyacetate.
Immediately underneath spacer layer 13 and in contact therewith is absorbent layer 14. The absorbent layer 14 is preferably formed from a cellulose material, e.g., absorbent cellulose paper.
Thus, the test device is comprised of layers 11, 12, 13 and 14, which are preferably combined to produce a unified device 10. The layers may be attached to each other, for example, by sewing of the layers to each other; however, other methods of attachment are possible.
As particularly shown, the test device 10 is in a test container which includes a base 15, and a cover 16. The base 15 has a depth such that the device 10 is within the container. In the preferred embodiment the container has three sides in a generally triangular shape with rounded corners.
The cover 16 includes a raised portion having a suitable aperture 17 which overlies the test area 18 of the solid support 11. Preferably, a portion of the background area of the support 11 surrounding the test area is also within the opening defined by aperature 17.
The cover 16 is supported over layer 11 by projections 19 extending upward from the sides of the base 15. The projections 19 are of sufficient height so as to provide air spaces 20 which provide for ventilation of the sides of the device 10. The air spaces 20 are bounded by the projections 19, the cover 16, and the base 15.
The raised portion of the cover 16 surrounding the aperture 17 include a colored area 21, the color of which preferably contrasts from that of the cover 16 and the color to be generated in test area 18 to provide for a better reading of the test results which are generally determined by color. In the preferred embodiment, base 15, cover 16, and colored area 21 are made of plastic materials.
For use of the test device 10 in a sandwich fluorescent signal when exposed to ultraviolet energy.
In manufacturing the test article, solid support 11 is initially treated with a mixture of binder and marker which may be a first fluorescent material to apply the mixture to test area 101. Thereafter, a mixture of the control and label which may be a second fluorescent material is applied to the control area 102. As shown, control area 102 is smaller and lies within area 101.
The fact that the analyte and binder have been deposited in their respective areas may then be determined by exposing solid support 11 to ultraviolet light and determining the presence of the respective fluorescent emission colors in the appropriate areas.
Thereafter, solid support 11 may be combined with the other layers, and the device 10 positioned in the test container in a manner such that the test area 101 is beneath aperture 17, whereby assay reagents may be directed to the area in which the binder and control is supported.
An alternative embodiment is shown in FIG. 4B wherein control area 102' is where the control is deposited and test area 101' is where the binder is deposited. As should be apparent, the presence of the control and the binder in their respective areas may be determined as described with reference to FIG. 4A.
In the embodiment of FIG. 4B, if no analyte is present, only area 102' is colored by the tracer, thereby indicating a negative sign on the layer 11. If analyte is present in the sample, assay, the binder may be an antibody specific for the analyte to be determined. If the sandwich assay is to be operated in a sequential mode a sample which is suspected of containing the analyte is applied to the test area through the aperture 17 whereby the sample contacts the binder in test area 18, with the sample flowing through the test device to the absorbent layer 14. The analyte present in the sample will become specifically bound to the binder in area 18.
Thereafter, tracer is applied to the test device through the aperture 17. The tracer becomes bound to the analyte, and any unbound portion flows through the test device to the absorbent layer 14.
If desired, a wash solution may be applied to the test device 10 prior to addition of the tracer. Similarly, after addition of the tracer, a wash solution may be applied to the test device to wash any tracer which may not be specifically bound to the complex in area 18, into the absorbent layer 14.
The presence of color in area 18 is indicative of the presence of analyte, and if the assay is to be a quantitative assay, the intensity of such color is indicative of the quantity of analyte present in the sample.
Referring now to FIG. 4A solid substrate 11 includes a test area 101 and control area 102. Test area 101 includes a mixture of the binder used in the assay and a marker, such as a fluorescent material. Control area 102 includes an analyte control in admixture with a label, preferably a second fluorescent material, which emits a both areas 101' and 102' will be colored, whereby a plus sign is displayed, indicating the presence of analyte in the sample.
Although the invention has been described with respect to a preferred embodiment as shown in the drawings, the present invention is equally applicable to depositing binder, and optionally also control on a solid support other than one particularly shown. Thus, for example, the test device may be in the form of a single layer, or two layers. Similarly, the test layer may be other than a membrane as shown.
These and other modifications should be apparent to those skilled in the art.
A sulfonyl chloride derivative of sulforhodamine 101 (available from Molecular Probes Cat. T353 as Texas Red Dye), after having been incubated for two hours in a 0.1 M glycine buffer pH 8.0 is combined with rabbit anti-group A Streptococcus antibody (n-acetyl glucosamine affinity purified) in a concentration ranging from 0.5 to 20 ug/ml. Separately Group A Streptococcus antigen is extracted with nitrous acid (300 ul HCl (0.1 M) with 40 ul NaNO2 (4M) and after three minutes 40 ul Tris buffer (1M, Trizma™ base (Sigma), with 4M NaCl)is added). To the extracted antigen is added Rhodamine 6G dye (Allied Cat. 663) in glycine buffer (0.1M) pH 8.0 to provide a concentration of less than 4 ug/ml.
Each of the above solutions is applied to nitrocellulose membrane (Schleicher & Schuell, pore size 5 microns) and under long wave (290-350 nm) ultraviolet excitation, the Texas Red, which fluoresces red, may be distinguished from the Rhodamine 6G which fluoresces bright yellow. The presence of the two colors discriminates and identifies the location of the antibody binder and antigen positive control.
The nitrocellulose containing the mixture of antibody and fluorescent material as well as the mixture of antigen positive control and fluorescent material is used in a sandwich assay for detecting Group A Strep antigen in which the tracer is affinity purified antibody (rabbit anti-group A Strep) covalently coupled to liposomes containing sulfo-rhodamine B dye (the procedure for making this tracer is described in South Africa Patent No. 84/9397 corresponding to allowed U.S. Ser. No. 579,667 which is hereby incorporated by reference).
The presence of the fluorescent material with the antibody binder and with the antigen positive control on the nitrocellulose support does not adversely affect sensitivity at 5×105 organisms/ml.
The present invention is applicable to procedures and products for determining a wide variety of analytes. As representative examples of types of analytes, there may be mentioned; drugs, including therapeutic drugs and drugs of abuse; hormones, vitamins, proteins, including antibodies of all classes; peptides; steroids; bacteria; fungi; viruses; parasites; components or products of bacteria, fungi, viruses, or parasites; allergens of all types; and products or components of normal or malignant cells. As particular examples, there may be mentioned T4 ; T3 ; digoxin; hCG; insulin; theophylline; luteinizing hormone; organisms causing or associated with various disease states, such as Streptococcus Pyogenes (group A), Herpes Simplex I and II, cytomegalovirus, rubella, chlamydia, and Candida Albicans.
The analyte may be determined in various samples, including, for example, body fluids, such as saliva, urine, serum, and cerebral spinal fluid or from swab samples, e.g., from the throat.
Numerous modifications and variations of the present invention are possible in light of the above, and therefore, within the scope of the appended claims, the invention may be practiced otherwise than as particularly described.
1. A test device for use in detecting the presence of an analyte, comprising:
a casing having an aperture;
a solid support comprising an upper surface and contained in said casing;
a test area on the upper surface of said support and positioned in registry with said aperture;
a binder for said analyte attached to said test area;
a detectable chromogen marker attached to said binder, related to the presence or absence of the binder which does not interfere with said assay; and
a background area on the upper surface of said support without binder and detectable marker;
wherein:
said marker causes the binder to be distinguishable from said support so that, before the device is used in said assay the device is examined to confirm that binder is present in said test area.
2. A test device for use in an assay for detecting the presence of an analyte, comprising:
a solid support comprising an upper surface and a lower surface;
a test area on the upper surface of said support;
a binder for the analyte attached to said test area;
a detectable marker attached to said binder which is related to the presence or absence of the binder, and does not interfere with said assay; and
a control area on the upper surface of said support;
an assay control attached to said control area; and
a detectable label attached to said assay control which is distinguishable and does not interfere with the assay or with the detectable marker;
the detectable marker causes the binder to be distinguishable from said support so that, before the device is used in said assay the device is examined to confirm that binder is present in said test area; and
said label causes the assay control to be distinguishable from said support so that before the device is used in said assay the device is examined to confirm that assay control is present on said support.
3. The test device of claim 2 wherein the assay control is a positive analyte control.
4. The test device of claim 3 wherein said control area is attached to said test area.
5. The test device of claim 2 wherein the label is a chromogen.
6. The test device of claim 5 wherein the solid support is nitrocellulose.
7. A process for producing a test device suitable for detecting the presence of an analyte, comprising:
providing a solid support comprising an upper surface;
contacting said upper surface with a binder for said analyte;
contacting said binder with a detectable marker related to the presence or absence of the binder which does not interfere with said assay;
detecting the marker to confirm that binder is present in said device; and
placing the support in a casing including an aperture with the test area in registry with the aperture.
8. The process of claim 7 further comprising:
contacting said upper surface with a mixture of an assay control; and
contacting said assay control with a detectable label;
said control area is distinguishable from said test area and said support, and said label is related to the presence or absence of said assay control.
9. The process of claim 8 wherein said control area is attached to said test area.
10. The process of claim 8 wherein the marker is a fluorescent material and the label is a different fluorescent material.
11. The process of claim 7 wherein the marker is a chromogen.
12. The process of claim 7 wherein the marker is a fluorescent dye.
13. The process of claim 7 wherein the support is nitrocellulose.
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To apply for one of the courses we provide, please click on the button below.
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The Transient High Energy Sky and Early Universe Surveyor (THESEUS) is a mission concept, under development by a large international collaboration and currently under evaluation by ESA within the selection process for next M5 mission, aimed at exploiting Gamma-Ray Bursts for investigating the early Universe and providing a substantial advancement of time-domain and multi-messenger astrophysics. These goals will be achieved through a unique combination of instruments allowing GRBs and X-ray transients detection over a broad FOV (more than 1sr) with 0.5-1 arcmin localization, an energy band extending from several MeVs down to 0.3 keV and unprecedented sensitivity in the soft X-ray domain, as well as on-board prompt (few minutes) follow-up with a 0.7m class IR telescope with both imaging and spectroscopic capabilities.
THESEUS is thus perfectely suited not only for addressing main open issues in cosmology such as, e.g., star formation rate and metallicity evolution of the ISM and IGM up to redshift 10, signatures of pop III stars, sources and physics of re-ionization, faint end of galaxies luminosity function, but also for performing an unprecedented monitoring of the X-ray variable sky, thus detecting, locating and identifying the electromagnetic counterparts to sources of gravitational radiation, which may be routinely detected in the late '20s / early '30s by next generation facilities like aLIGO/aVirgo, eLISA, KAGRA, Einstein Telescope, and providing a perfect service and sinergy also to next generation multi-wavalength (e.g., E-ELT, SKA, CTA, ATHENA) observatories.
The aim of this Workshop is to collect the several astorphysical communities involved and interested in the scientific goals, and related technology, of THESEUS, in order to review the status of the project and further discuss and refine the expected scientific return for the several fields of cosmology and astrophysics on which this mission would have an important impact.


"Nobody does Dylan like Dylan,'' indie folk maestro Josh Pyke told a Brisbane audience last night before adding "especially my father''.
His father would wander around the family home and sing Tangled Up in Blue – a childhood memory that didn't diminish Pyke's love of that Bob Dylan classic.
Even professional, accomplished musicians have butchered Dylan songs. I would sooner bleed from the ears than listen to Olivia Newton-John sweetly sing If Not For You.
Others attempt to imitate Dylan, becoming poor clones of the real deal. A few manage to reinterpret and arguably improve on the originals. By many accounts, Dylan was so enamoured with Jimi Hendrix's version of All Along the Watchtower that he took to performing the late, great guitarist's version of the anthemic tune.
The master ... Bob Dylan.
So it was with trepidation and excitement in equal measure that I headed off last night to the Lyric Theatre for Bob Dylan Night: The 50th Anniversary Concert.
Pyke shared the stage – shrouded in smoke and bathed in retina-singeing lights - with Eskimo Joe's Kav Temperley, Jebediah's Kevin Mitchell, Holly Throsby, and The Grates' Patience Hodgson.
It was a rollicking good time despite the venue being conducive to sitting while politely clapping and nodding in time with the music rather than surrendering to the urge to jump to your feet and dance, particularly when Temperley tackled All Along The Watchtower and Rainy Day Women #12 & 35 as Hodgson and Mitchell commandeered tambourines and danced through the aisles like hipsta Hare Krishnas.
The band was brilliant and the playlist would please music-lovers who had enjoyed the most casual fling with Dylan.
It's All Over Now Baby Blue, Blowin' In the Wind, Hurricane, The Times They Are A-changin', Positively 4th Street, Lay Lady Lay, Just Like A Woman and Subterranean Homesick Blues were among the popular songs that made the cut.
One highlight was Temperley, Mitchell and Pyke's rendition of Mr Tambourine Man, but it was more The Bryds than Dylan. Mitchell and Hodgson teamed up to perform If Not For You – a version stripped of its angst and transformed into a pop song.
Throsby's breathy and haunting interpretation of Girl From the North Country was sublime as was Hodgson's version of Visions of Johanna and Temperley's rendition of It Aint Me Babe, with the Eskimo Joe frontman remarking it felt like he had written the song.
The only left-field selection was Tomorrow Is A Long Time sung by Throsby who had liked the song after hearing it on Masterpieces – a 3-LP compilation that she has owned since high school.
The talented quintet finished the two-and-a-half-hour concert with a crowd-pleasing encore, singing Like A Rolling Stone as the audience was liberated from their seats and invited to dance.
But Jimi "sang them exactly the way they were intended to be sung and he played them the same way. He did them the way I would've done them if I was him,'' he said according to an article on Jimi and Bob Dylan: A Cosmic Friendship.
At the Lyric Theatre last night, I doubt there was a Hendrix experience but it was a thoroughly entertaining romp through the substantial back catalogue of the one of modern history's greatest musicians.


text:
Harley Davidson Ride Atlas of North America
Earlier this month, Rand McNally introduced a new road map for motorcycle riders, dubbed " Harley Davidson Ride Atlas of North America ...
https://www.bikernewsonline.com/2006/05/harley-davidson-ride-atlas-of-north.htm
Earlier this month, Rand McNally introduced a new road map for motorcycle riders, dubbed "Harley Davidson Ride Atlas of North America ".
It's packed with definitive maps of secondary roads, road construction information, local motorcycle laws and an expansive mileage directory, into a compact 8.5-inch by 11-inch ride atlas fits snugly into a saddlebag.
Wrapped in a black, rubber-like, touche cover are 284 pages of four-color maps, mileage charts, city indexes and descriptions of motorcyclist-tested rides in the United States.
The Harley-Davidson Ride Atlas is packed with in-depth descriptions, custom trip maps and essential rider tips for 25 key rides nationwide. The scenic rides take motorcyclists off the beaten path -- to the coves, hollows and water gaps of North Carolina's Blue Ridge Parkway, through the towering oaks and cedars of Texas Hill Country, and past the emerald lakes and swift streams of the Cascade Range in northern California.
Other unforgettable rides climb from the southern tip of Lake Superior toward the Canadian border along state Route 61, or hug the shoulders along the Chief Joseph Scenic Byway in Wyoming. Others meander through history along "The Trace" in the Land Between the Lakes in Kentucky and Tennessee. Still others cruise down a desert highway well-stocked with gas stations -- but no roadwork -- or crisscross the Rio Grande along the U.S./Mexico border.
Each ride features its own map highlighting the route, nearby points of interest and geographic features. The rides also include useful touring information, suggested side trips, best seasons to ride, gasoline availability, road conditions, Web sites and phone numbers for state tourism resources, and other insider tips.
In addition to ride maps, the Ride Atlas contains reliable, detailed Rand McNally maps of each U.S. state, major U.S. cities, Canada, Mexico and Puerto Rico. The maps are tailored to riders' needs, with the minor highways and secondary roads highlighted in bold color. The 25 featured rides and other scenic routes also are highlighted brightly.
Each state map features listings, locations and phone numbers of Harley-Davidson dealerships so that you won't miss getting that dealer t-shirt.
Beside the maps are crucial state motorcycle laws and contacts for emergency roadside assistance, state motor vehicle information and tourism resources.
The Ride Atlas also features detailed border crossing information for Mexico and Canada, contacts for U.S. state parks and profiles of America's 10 most popular national parks. A mileage and minutes guide provides driving distances and times between major U.S. cities. It also includes information on Harley-Davidson factory tours, Rider's Edge -- The Harley-Davidson Academy of Motorcycling, Harley-Davidson Authorized Rentals, the Harley Owners Group and motorcycle shipping.
The Ride Atlas is available now at authorized Harley-Davidson dealerships, Randmcnally.com, select bookstores, select automotive stores, truck stops and military outlets.
Suggested retail price is $34.95.
Maps 114853601253518730

redpajama_set_name: RedPajamaCommonCrawl


Author Kathryn Mattingly brings us her new title, The Tutor, in the coming months. Kathryn is a gifted writer with a growing catalog. Catch up on her previous titles available in print and eBook.
This May we will be releasing Prismatic, a young-adult fantasy but author Sarah Elle Emm. Prismatic follows a group of segregated inter-racial youths who must work to find acceptance and freedom.
Welcome the very talented Gary Beck!


This Autumn Pafuri Camp is offering a special rate R3,170.00 per person sharing per night. The package includes, accommodation, all meals, two game drives per day and a bottle of wine per booking. It excludes all travel costs, gratuities, park entry fees, conservation levies and beverages.
Pafuri Camp has something to offer everyone, from nature fundis to the easy going safari holidayer. With the options to either relax on your private viewing deck from your room, go on a game drive and learn about the wildlife, ecology and bird life of Pafuri, to stopping off at look out points to see Pafuri from differing vantages, or to enjoy something a little more challenging like our bush walks. These walks take you along trails that the animals have walked for centuries, letting you experience nature at its best and off the beaten track.
At Pafuri Camp we want everyone to go at their own pace. Be it watching the sunrise in bed, relaxing besides the pool in the afternoon or taking a stroll along the raised wooden walkways connecting the camp.
Each tent is set apart from the others, offering privacy and your own sanctuary. Each luxury tent has an outlook of the Luvuvhu river, where you can watch the hippos and crocodiles at play will enjoying your morning coffee.


When is the last time you did absolutely nothing for 10 whole minutes? Not texting, talking or even thinking?
In this part of his talk, Bill White discusses the degrees/depths of recovery. He describes how some better people feel 'better than well' after recovery.
He goes on to describe different types of recovery initiation/maintenance framework and different styles of recovery.
Danielle Tarino is a person in long-term recovery from alcohol and other drugs since age 19. Danielle graduated from Rutgers University as a member of the Rutgers Recovery House on campus with a degree in political science and psychology.
Resulting from her own personal experience graduating from a supportive collegiate recovery program, Danielle has harnessed the opportunity to bring the lived youth recovery voice to Washington, D.C. serving as a Public Health Advisor for the federal government's Substance Abuse and Mental Health Services Administration.
Marion believes her culture is changing and she has learned to adapt when changes occur.
"Here's to the crazy ones. The misfits. The rebels. The troublemakers. The round pegs in the square hole. The ones who see things differently.
Bill describes a metaphor for explaining a paradigm shift, such as the paradigm shift to recovery-based care in mental health.
This metaphor is that mankind once thought the world was flat. This understanding led to certain questions such as, "How far do I sail before I fall off the end of the earth?" Once mankind learnt the world was round, these questions were redundant. We asked different questions.
Please check out Jim's blog. Jim is the executive director of a counseling center in the States that specialises in addiction, dual diagnosis, and trauma.
Barwon Health Drug and Alcohol Services, together with Salvation Army Kardinia Alcohol and Other Drugs Service, facilitate 'Making Changes' – an ongoing therapeutic recovery group in Geelong, Victoria, for people healing from alcohol and other drug addictions.
Art Therapy is used as part of the therapeutic processes of the group. These processes provide insight into the difficult journey individuals undertake on the road to recovery.
To recognise National Drug Action Week 2013, clients consented to sharing their recovery through a public display expressing their stories. An Exhibition entitled 'Recovery Journeys Expressed' was held at Deakin University, Waterfront Campus, Geelong during Drug Action Week – 17th to 21st of June.
In my last blog, I referred to Stephanie Brown's book A Place Called Self, in which she describes recovery as radical change in personal identity, or the self. Stephanie goes on to emphasise a number of myths about recovery.
Stephanie emphasises that recovery is more like a starting over than a restoration of what was lost during addiction. This is because for many people the real self was never really developed.


text:
Q: $(window).scroll() Loop, due to no variable I have a bit of a problem; I have this code:
  $(window).scroll(function(){
    if ($(document).scrollTop() >= $('.services-container').offset().top-80) {
      alert("test");
    }
  });

Now everything works ok, except I need to run the if statement once, I tried variables but I cannot get them not to re-set to previous state due to the window scroll loop... any help?

A: You can try with some flag:
var flag = true;

$(window).scroll(function() {
    if ($(document).scrollTop() >= $('.services-container').offset().top-80 && flag) {
       alert("test");
       flag = false;
    }
});

Demo Fiddle

A: Try this:
var flag = true;
$(window).scroll(
    function() {
        if ($(document).scrollTop() >= $('.services-container').offset().top-80 && flag) {
            flag = false;
            alert("test");
        }
    }
);

A: You can use;
var enabled = true;
$(window).scroll(
    function() {
        if ($(document).scrollTop() >= $('.services-container').offset().top-80 && enabled) {
            alert("test");
            enabled = false;
        }
    }
);

Here is working jsFiddle.

redpajama_set_name: RedPajamaStackExchange


I was having lunch last week with a workout officer at a large bank. He was commenting on how customers referred to his department were often in a state of denial. Not unlike the man who finds himself in Hell but won't accept it!
It seems that a lot of businesses are in denial. Their sales are off on average thirty percent, they have been collecting old receivables and are losing money. Furthermore, they know they are going to have a problem with their debt compliance certificate in the coming quarter. Yet, they wait for something to change.
It is important for all of us to accept that this recession will last longer that those in the past. It already is one of the longest!
One of the perplexities of finance is that a company often has more cash when it is shrinking and going out of business than when it is growing. Consequently, a number of companies are generating cash and losing money at the same time. In a way they are consuming their flesh! At some point they will have run through their liquidity and have fewer options to save their company.
What every company should be doing is developing a business plan to reposition themselves to first stabilize the profitability at a lower level of sales and, more importantly, take advantage of the changes taking place. It all starts with making a business plan and putting it in writing.


August 2018 – blindambitionsgroups.org | We may have lost our sight, but we have not lost our VISION!
One of the primary necessary life skills, math opens many possibilities in the kid's adult life and while we are aware that it is a must, a mind may not be made to comprehend. The Expanded Core Curriculum, designed to produce general curriculum accessible for students that are blind or have a visual impairment, deems a student can just have success in the overall curriculum should they have data in a format that's accessible to their precise needs. It's critically important that we're giving students that are blind or visually impaired the skills they will need to attain their fullest potential. 1 student, who's equipped to observe colors at a brief distance, couldn't quit laughing whenever the color names the screen reader gave matched the colors he managed to see. Special education was typically only provided in big cities. Students may fall over things they aren't utilized to being in a sure spot. Taking a peek at the legal history shows precisely how much progress was made in educating students with disabilities in the USA, and how much more is needed.
At Purdue, there's a program called EPICS in which students may work with real world clients to engineer products to be applied locally at large. It's simple to think of projects where empathy was not used. Then you warn the men and women in your community, and seek out a company that's prepared to fulfill your wants.
A responsive layout will enable your design to scale at different screen sizes on various devices. The plan should react to the user and their requirements. Prototyping responsive design is in fact pretty easy in Justinmind as a result of our latest update. Seems like some designers aren't conscious of the Photoshop built-in feature that permits you to preview the design among the two most-common groups of color blinds.
Contrast isn't pretty much color though. As an example, providing sufficient contrast between your site's foreground elements and background elements may look like a minor detail, but nevertheless, it can be a large improvement for users which are color blind or have difficulty seeing. Color blind folks gain from well-contrasting colours. You may refer to it should you truly feel like you would like to know more of my background.
Specialized technologies make it possible for teachers to customize instruction when giving more time to the whole class, and offer different students with an enriching educational experience. Laptop computers with readers ought to be supplied to visually impaired students, together with keyboarding instruction. Back then, there were not any personal computers, no web, zero smartphones. Despite the fact that it can be utilized with the keyboard, it doesn't report plenty of things.
What Tools Are There For Visually Impaired Students: the Ultimate Convenience!
Both have smartphone apps too. The app changes the colours of whatever you see on the screen according to the sort of color-blindness you decide on. A good deal of folks use various applications to visualize things. To begin with, try to earn your web or app distinguishable. Although most users do require some training, the majority of people can walk up and get started employing a SMARTBoard immediately. You will soon understand a number of the pains users with disabilities go through when seeking to use a cell device. Likewise in code editors the majority of the feedback is visual and can't be understood through screen readers.
In chronic leukemia, early indications and symptoms might not be apparent for months. The indications of acromegaly aren't only huge hands and feet and significant height. Anyway, if your symptoms have ceased, it's no indication you could start the use of alcohol all over again. Signs of inner ear issues can be debilitating, and since the signs are so varied, it is quite often misdiagnosed.
In some instances, the headache may worsen, the moment the kid is lying down, or whenever the child coughs or sneezes. It is supposed to be one of the most common chronic complaints of people all around the world. Tension headaches are among the most frequent types of headaches. My pain of a busted arm and a dislocated shoulder couldn't be relieved with pain pills on each occasion, on account of the allergic reactions that I now had when taking brain altering pharmaceuticals.
The signs of diabetes are similar for both women and men. They should not be neglected, if the headaches worsen. The signs of inner ear disorders shouldn't be ignored, as some conditions can cause permanent hearing loss. The common symptoms incorporate Many members of the world test for diabetes only when they find wounds that aren't healed for a very long time in their foot. The indications of leukemia will differ based on its type. Acute leukemia symptoms are apparent within a brief time period.
The ear is just one of the most vital sensory organs of the body. You will also learn to safeguard your eyes when using technology and screens. Eyesight might no longer continue being normal in the event the brain function was damaged considerably.
There's marked rise in the frequency of urination. According to some studies, it is thought that honey have laxative effects only owing to its incomplete absorption within the body. It's further imperative to comprehend the side effects of drugs, particularly when they should be given to children.
Certain medications like benzodiazepine drugs are beneficial in treating inner ear troubles. The dosage of the drug needs to be increased, or a different corticosteroid drug should be prescribed to control the signs of the status. Prednisone is a kind of steroid which helps restore immunity levels, which malfunction for a consequence of illness or injury. It comes under the category of drugs called corticosteroids.
If you're taking blood-thinning medications or anticoagulants like aspirin, then do check in with your physician to figure out if it's safe that you consume papaya. Fully being a low-cost and efficient drug, it's often prescribed by doctors. Only a health care provider would have the capacity to determine the right plan of treatment. In addition, the physician may suggest the crucial supplements that will need to get taken orally to overcome riboflavin deficiency. Doctors haven't been in a position to recognize the precise causes of leukemia. They advise their patients not to consume alcohol, especially in the first phase of therapy when the possible side effects of prednisone have to be understood. Besides, in the event the patient has been a drinker, the physician will prescribe a few tests to establish the level of liver damage, if any, as a result of excess alcohol consumption.


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Apple Reportedly Working on Redesigned Apple TV Remote
Tuesday March 30, 2021 1:23 am PDT by Sami Fathi
Rumors that Apple intends to launch a new Apple TV have been around for some time, with the headlining feature being a more powerful processor for gaming and all-round performance benefits. However, alongside a new powerful chip, new evidence is pointing towards a possible new Apple TV remote also shipping alongside the TV set-top box.
According to a report from 9to5Mac, citing people familiar with Apple's plans, the company has been developing a new Apple TV remote under the codename "B519," which differs from the current "B439" codename for the existing remote. Specifics on what to expect from the new remote aren't provided; however, 9to5Mac believes the changes will be "significant."
The last time Apple updated the remote was with the Apple TV 4K in 2017, when the company added a white border around the Menu button. However, the remote has remained unchanged since, while its slimline design and lack of physical navigation buttons continues to divide users.
MacRumors recently discovered that within the tvOS 14.5 beta, Apple has renamed the remote from "Siri Remote" to "Apple TV Remote" in multiple instances across the system. The system-wide change in wording could indicate that the new remote will have less of a focus on Siri and instead be designed around more traditional content navigation and other TV functionality.
Related Roundup: Apple TV
Buyer's Guide: Apple TV (Neutral)
Related Forum: Apple TV and Home Theater
MacFather
x5tuu
I really don't get the hate for the remote at all - we have 4 apple TVs in the house all with the touch remote and everyone uses them without any problem at all (including a tech-averse wife and our 6yr old) and for most of those rooms they have become the primary remote that is used. The remote is possibly the simplest piece of 'just works' tech that I have seen in a long while.
LeeTom
Thank god. Either the touch is half baked in hardware or it just doesn't work. Would love an all-button remote.
Just as long as it has a Mute Button.
Juicy Box
I don't understand why anyone even uses the remote. Not becuse it's bad but because your phone is almost always at hand, not lost down the back of the sofa and just seems like an easier alternative.
At least for me, the Remote app on the iPhone takes most of the bad stuff about the Siri Remote such as lack of physical buttons, a sudo D-Pad, no mute, and then double downs on it with no physical buttons, no D-pad, and no volume buttons.
Basically making the experience on the Remote app much worse than so-so experience with the Siri Remote.
Plus.......
your phone is almost always at hand
Mine is not.
When I am home, I hardly touch my iPhone. At home, I much rather use a Mac or ATV for anything I would normally do with an iPhone while not at home.
BMox81
No issues with the remote in our house. My daughter loves using it now.
Apple TV 4K Very Easy to Repair but Siri Remote Battery Difficult to Replace, Teardown Shows
Wednesday May 26, 2021 5:00 am PDT by Hartley Charlton
iFixit has released its teardown of the second-generation Apple TV 4K, revealing an easy to repair, modular design, alongside a laborious process to dismantle the redesigned Siri Remote. As with previous Apple TV models, the entire plastic shell of the device is transparent to IR light, allowing the Siri Remote to be used from any angle. The large fan inside the Apple TV is not connected...
Apple's TV Remote App Pulled From App Store Since Functionality is Available in Control Center
Tuesday October 20, 2020 10:26 pm PDT by Juli Clover
Apple recently pulled the "Apple TV Remote" app from the App Store, which was designed to let users control their Apple TVs from their iPhones or iPads with a touch-based remote interface. It's not quite clear when Apple nixed the app, but it was quietly removed and clicking on the prior link for the app confirms that it is no longer available. 9to5Mac discovered that Apple has also removed...
New Apple TV Ships With Redesigned Siri Remote, Also Sold Separately for $59
Tuesday April 20, 2021 10:19 am PDT by Joe Rossignol
Apple today announced the next-generation Apple TV 4K with a faster A12 chip and a completely redesigned Siri Remote pictured below. The new Siri Remote is included with the new Apple TV 4K, and the Apple TV HD going forward. The new remote will also be sold separately for $59 and is compatible with the previous-generation Apple TV 4K and Apple TV HD. The all-new Siri Remote features a...
New Siri Remote Doesn't Have U1 Chip and Won't Work With Find My
Tuesday April 20, 2021 4:06 pm PDT by Juli Clover
The newly redesigned Siri Remote, which will be sold alongside the updated Apple TV 4K and is also available on a standalone basis, does not have a built-in U1 chip and does not appear to be compatible with the Find My app. There is no U1 ultra wideband chip listed in the specs for the Siri Remote, nor is there a mention of the Find My app, and now Apple has confirmed to The Verge that it's...
Hands-On With the Redesigned Apple TV 4K Siri Remote
Monday May 24, 2021 12:20 pm PDT by Juli Clover
Apple alongside the second-generation Apple TV 4K debuted a redesigned version of the Siri Remote, which is now available to customers with new Apple TV orders and as a $60 standalone replacement for the original Siri Remote. Subscribe to the MacRumors YouTube channel for more videos. We picked up the updated Siri Remote to see if it's worth picking up if you have an original Apple TV 4K or...
Universal Electronics Offers Alternative Apple TV Remote to Cable Companies
Sunday November 22, 2020 2:02 am PST by Tim Hardwick
Universal Electronics has announced its own take on Apple's divisive Apple TV Remote that's "specifically developers and designed" for cable, satellite, and MVPD (multichannel video program distributor) customers, according to the company (via The Verge). The UEI Remote uses Bluetooth Low Energy (BLE) connectivity together with Infrared to provide universal control, and offers Apple TV users ...
tvOS 14.5 Beta 6 References Apple TV Remote With 'Center Button' [Updated]
Wednesday March 31, 2021 2:06 pm PDT by Juli Clover
The tvOS 14.5 beta that was released this morning includes a code change that references an Apple TV remote control that features a "Center Button," which is a description that does not fit the current Apple TV remote. "Press the center button or the touch surface to continue," reads the tvOS 14.5 code, which was discovered by MacRumors contributor Steve Moser. The remote for the fourth...
New Siri Remote Lacks Accelerometer and Gyroscope for Gaming on Apple TV
Thursday April 22, 2021 3:48 pm PDT by Joe Rossignol
The new Apple TV 4K unveiled this week ships with a redesigned Siri Remote with a physical clickpad, new power and mute buttons for a TV, and a repositioned Siri button, but there's another change that has gone less noticed. According to Digital Trends, the new Siri Remote lacks an accelerometer and gyroscope, which allowed the original Siri Remote to function as a gaming controller. Apple...


Activities are one of the most important parts of a college application. They show colleges that students are well-rounded and don't only focus on grades. Not sure which ones you should list? Read on for some ideas.
Include any groups you are involved with in high school. You can list larger organizations such as student government or the student newspaper. You can also put down any smaller clubs you are involved that are based around shared hobbies or culture. Another great thing to add is involvement in competitive academic teams such as debate, MUN, or mock trial.
Involvement in sports teams shows both leadership and teamwork skills. You can list sports whether you play on a team inside or outside of school. You can also list if you train privately in an individual sport such as tennis. Solo sports still show that you are committed and hardworking.
Do you read to kids at the library or volunteer at a homeless shelter? These are great activities to include in a college application. If you show that you were a productive, generous member of your home community, colleges will believe that you will also be able to contribute in the same way to their school community.
You should definitely list activities like theater, dance, orchestra, music lessons, and visual arts, whether you do them inside or outside of school. Even if you don't plan on majoring in an artistic field, you will still be able to contribute to the cultural community of any college you attend. Every school wants their campus to be an artistically vibrant place, so show them that you can help create that atmosphere.
Start practicing resume building now. No matter where you work, it will look good on a college application. This includes jobs in retail and fast food. Babysitting can also count if it's something you're committed to and do very frequently.
Don't list any chores as activities on your application. Colleges don't care if you wash the dishes every night. However, if you spend a substantial amount of time caring for an older relative or helping your family out in a significant way, then definitely put it in your application. You'll show that you are responsible, organized and mature.
It's better to have a few impressive activities on your application than a dozen random clubs that you didn't really contribute to. Try to think about how you spent most of your time in high school, what you care about the most, and what you love the most. These activities will show admissions officers who you truly are.
Worried about how you'll balance all of those applications with your homework and extracurriculars? Read this post for some tips.
Want a well-rounded college application? Join the club.


At the currently on-going Perfect World event, CEO Robert Hong Xiao has given the word that the first beta wave of the Chinese Dota 2 servers will be coming later this month, on April 28.
According to the details, this will mark the end of Dota 2 via the Steam platform in China — it is yet to be seen whether Chinese players will have free interactions with international players (we hope that everything remains open).
Update: Chinese Dota 2 will run on a stripped down version of Steam, re-done specifically for Dota 2 in China. Inter-connectivity between players around the world will remain!
Pingback: Perfect World's Dota 2 league coming in May | Dota 2 CN -- Dotaland!


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Q: Use name of controllers to find table name in Select I want to extract a ID , User_ID value from one of the Companies and Contract tables, depending on the ContorollerName value.
select P.TitleProject, P.StartDateProject, P.EndDateProject,P.ControllerID,P.RecordID,P.IsAllocated,P.ProjectStatus_ID,
CN.ControllerName,CN.PersianName,
PU.ProjectID,PU.UserID,PU.RoleID,
    CASE
        WHEN CN.ControllerName = 'Company' THEN
            Companies.Id,Companies.[User_Id]
        WHEN CN.ControllerName = 'Contract' THEN
            Contracts.Id,Contracts.[User_Id]
        END
from Projects P
    left outer join Controllers CN ON P.ControllerID = CN.Id
    left outer join ProjectUsers PU ON P.Id = PU.ProjectID
where P.IsAllocated = 1

For example, if ContorollerName is 'Company' , the select command is as follows :
select P.TitleProject, P.StartDateProject, P.EndDateProject,P.ControllerID,P.RecordID,P.IsAllocated,P.ProjectStatus_ID,
CN.ControllerName,CN.PersianName,
PU.ProjectID,PU.UserID,PU.RoleID,
Companies.Id,Companies.[User_Id]

A: You are on the right track -- using left join.  But you need to add the tables to the from clause with the appropriate logic.
The logic for the join is quite unclear.  The query looks something like this:
select . . .,
       coalesce(c.id, co.id) as id,
       coalesce(c.user_id, co.user_id) as user_id
from Projects P left join
     Controllers CN
     on P.ControllerID = CN.Id left join
     ProjectUsers PU
     on P.Id = PU.ProjectID left join
     companies c
     on c.? = ? and   -- no idea what the right join conditions are
        c.ControllerName = 'Company' left join
     contracts co
     on co.? = ? and  -- no idea what the right join conditions are
        co.ControllerName = 'Contract'
where P.IsAllocated = 1