| 1. |
- Aifa, Sami, 1967-, et al.
(författare)
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A basic peptide within the juxtamembrane region is required for EGF receptor dimerization
- 2005
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Ingår i: Experimental Cell Research. - : Elsevier BV. - 0014-4827 .- 1090-2422. ; 302:1, s. 108-114
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Tidskriftsartikel (refereegranskat)abstract
- The epidermal growth factor receptor (EGFR) is fundamental for normal cell growth and organ development, but has also been implicated in various pathologies, notably tumors of epithelial origin. We have previously shown that the initial 13 amino acids (P13) within the intracellular juxtamembrane region (R645-R657) are involved in the interaction with calmodulin, thus indicating an important role for this region in EGFR function. Here we show that P13 is required for proper dimerization of the receptor. We expressed either the intracellular domain of EGFR (TKJM) or the intracellular domain lacking P13 (ΔTKJM) in COS-7 cells that express endogenous EGFR. Only TKJM was immunoprecipitated with an antibody directed against the extracellular part of EGFR, and only TKJM was tyrosine phosphorylated by endogenous EGFR. Using SK-N-MC cells, which do not express endogenous EGFR, that were stably transfected with either wild-type EGFR or recombinant full-length EGFR lacking P13 demonstrated that P13 is required for appropriate receptor dimerization. Furthermore, mutant EGFR lacking P13 failed to be autophosphorylated. P13 is rich in basic amino acids and in silico modeling of the EGFR in conjunction with our results suggests a novel role for the juxtamembrane domain (JM) of EGFR in mediating intracellular dimerization and thus receptor kinase activation and function. © 2004 Elsevier Inc. All rights reserved.
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| 2. |
- Aifa, Sami, et al.
(författare)
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Phosphorylation of Thr654 but not Thr669 within the juxtamembrane domain of the EGF receptor inhibits calmodulin binding
- 2006
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Ingår i: Biochemical and Biophysical Research Communications - BBRC. - : Elsevier BV. - 0006-291X .- 1090-2104. ; 347:2, s. 381-387
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Tidskriftsartikel (refereegranskat)abstract
- Calcium-calmodulin (CaM) binding to the epidermal growth factor receptor (EGFR) has been shown to both inhibit and stimulate receptor activity. CaM binds to the intracellular juxtamembrane (JM) domain (Met645-Phe688) of EGFR. Protein kinase C (PKC) mediated phosphorylation of Thr654 occurs within this domain. CaM binding to the JM domain inhibits PKC phosphorylation and conversely PKC mediated phosphorylation of Thr654 or Glu substitution of Thr654 inhibits CaM binding. A second threonine residue (Thr669) within the JM domain is phosphorylated by the mitogen-activated protein kinase (MAPK). Previous results have shown that CaM interferes with EGFR-induced MAPK activation. If and how phosphorylation of Thr669 affects CaM-EGFR interaction is however not known.In the present study we have used surface plasmon resonance (BIAcore) to study the influence of Thr669 phosphorylation on real time interactions between the intracellular juxtamembrane (JM) domain of EGFR and CaM. The EGFR-JM was expressed as GST fusion proteins in Escherichia coli and phosphorylation was mimicked by generating Glu substitutions of either Thr654 or Thr669. Purified proteins were coupled to immobilized anti-GST antibodies at the sensor surface and increasing concentration of CaM was applied. When mutating Thr654 to Glu654 no specific CaM binding could be detected. However, neither single substitutions of Thr669 (Gly669 or Glu669) nor double mutants Gly654/Gly669 or Gly654/Glu669 influenced the binding of CaM to the EGFR-JM. This clearly shows that PKC may regulate EGF-mediated CaM signalling through phosphorylation of Thr654 whereas phosphorylation of Thr669 seems to play a CaM independent regulatory role. The role of both residues in the EGFR-calmodulin interaction was also studied in silico. Our modelling work supports a scenario where Thr654 from the JM domain interacts with Glu120 in the calmodulin molecule. Phosphorylation of Thr654 or Glu654 substitution creates a repulsive electrostatic force that would diminish CaM binding to the JM domain. These results are in line with the Biacore experiments showing a weak binding of the CaM to the JM domain with Thr654 mutated to Glu. Furthermore, these results provide a hypothesis to how CaM binding to EGFR might both positively and negatively interfere with EGFR-activity. © 2006 Elsevier Inc. All rights reserved.
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| 8. |
- Alimelli, Adriano, et al.
(författare)
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Fish freshness detection by a computer screen photoassisted based gas sensor array
- 2007
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Ingår i: Analytica Chimica Acta. - : Elsevier BV. - 0003-2670 .- 1873-4324. ; 582:2, s. 320-328
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Tidskriftsartikel (refereegranskat)abstract
- In the last years a large number of different measurement methodologies were applied to measure the freshness of fishes. Among them the connection between freshness and headspace composition has been considered by gas chromatographic analysis and from the last two decades by a number of sensors and biosensors aimed at measuring some characteristic indicators (usually amines). More recently also the so-called artificial olfaction systems gathering together many non-specific sensors have shown a certain capability to transduce the global composition of the fish headspace capturing the differences between fresh and spoiled products. One of the main objectives related to the introduction of sensor systems with respect to the analytical methods is the claimed possibility to distribute the freshness control since sensors are expected to be "portable" and "simple". In spite of these objectives, until now sensor systems did not result in any tool that may be broadly distributed. In this paper, we present a chemical sensor array where the optical features of layers of chemicals, sensitive to volatile compounds typical of spoilage processes in fish, are interrogated by a very simple platform based on a computer screen and a web cam. An array of metalloporphyrins is here used to classify fillets of thawed fishes according to their storage days and to monitor the spoilage in filleted anchovies for a time of 8 h. Results indicate a complete identification of the storage days of thawed fillets and a determination of the storage time of anchovies held at room temperature with a root mean square error of validation of about 30 min. The optical system produces a sort of spectral fingerprint containing information about both the absorbance and the emission of the sensitive layer. The system here illustrated, based on computer peripherals, can be easily scaled to any device endowed with a programmable screen and a camera such as cellular phones offering for the first time the possibility to fulfil the sensor expectation of diffused and efficient analytical capabilities. © 2006 Elsevier B.V. All rights reserved.
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| 9. |
- Anderson, Tony, 1973-, et al.
(författare)
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Frog melanophores cultured on fluorescent microbeads : Biomimic-based biosensing
- 2005
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Ingår i: Biosensors & bioelectronics. - : Elsevier BV. - 0956-5663 .- 1873-4235. ; 21:1, s. 111-120
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Tidskriftsartikel (refereegranskat)abstract
- Melanophores are pigmented cells in lower vertebrates capable of quick color changes and thereby suitable as whole cell biosensors. In the frog dermis skin layer, the large and dark pigmented melanophore surrounds a core of other pigmented cells. Upon hormonal stimulation the black-brown pigment organelles will redistribute within the melanophore, and thereby cover or uncover the core, making complex color changes possible in the dermis. Previously, melanophores have only been cultured on flat surfaces. Here we mimic the three dimensional biological geometry in the frog dermis by culturing melanophores on fluorescent plastic microbeads. To demonstrate biosensing we use the hormones melatonin and α-melanocyte stimulating hormone (α-MSH) as lightening or darkening stimuli, respectively. Cellular responses were successfully demonstrated on single cell level by fluorescence microscopy, and in cell suspension by a fluorescence microplate reader and a previously demonstrated computer screen photo-assisted technique. The demonstrated principle is the first step towards "single well/multiple read-out" biosensor arrays based on suspensions of different selective-responding melanophores, each cultured on microbeads with distinctive spectral characteristics. By applying small amount of a clinical sample, or a candidate substance in early drug screening, to a single well containing combinations of melanophores on beads, multiple parameter read-outs will be possible. © 2004 Elsevier B.V. All rights reserved.
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| 12. |
- Arwin, Hans, 1950-, et al.
(författare)
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Computer screen photo-assisted measurement of intensity or polarization change of light upon interaction with a sample
- 2006
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Patent (populärvet., debatt m.m.)abstract
- A method for measuring a property of a test sample utilizing one of the test methods ellipsometry, surface plasmon resonance and nephelometry, wherein the method includes the steps; providing a test sample being an object with which an optical interaction with light takes place, illuminating said test sample using a program controlled display as a light source, which program controlled display is composed of at least one activated pixel providing an illumination from an illuminating area of said program controlled display, arranging said program controlled display to illuminate said test sample with polarized light, detecting light emerging from said test sample utilizing a detector coupled to said program and evaluating said property from signals from said detector.
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| 15. |
- Bakker, Jimmy W.P., et al.
(författare)
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Immunodetection using computer screen photo-assisted ellipsometry
- 2008
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Ingår i: Physica Status Solidi. C: Current Topics in Solid State Physics. - Weinheim, Germany : Wiley-VCH Verlagsgesellschaft. - 1862-6351. ; 5:5, s. 1431-1433
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Tidskriftsartikel (refereegranskat)abstract
- Detection of antibody-antigen reactions is demonstrated by measuring changes in reflectance of light polarized parallel to the plane of incidence, using a computer screen as light source and a web camera as detector, giving results similar to traditional off-null ellipsometry and in accordance with a simplified theoretical model.
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| 16. |
- Barkå, Jonas, et al.
(författare)
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Background compensation in computer screen photo-assisted reflectance fingerprinting
- 2006
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Ingår i: Sensors and actuators. B, Chemical. - : Elsevier BV. - 0925-4005 .- 1873-3077. ; 120:1, s. 79-85
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Tidskriftsartikel (refereegranskat)abstract
- The computer screen photo-assisted technique (CSPT) is a method for the classification of colorimetric assays utilizing ordinary computer sets and web cameras as instrumentation. In CSPT measurements the web camera captures the image of the assay under the screen illumination, and typically a spurious spatial distribution of intensities is overlapped on the image. This issue is examined here, focusing on the effect of the sample and illuminating colors on the spatial modulation of intensity. A method for the selection of colors composing an illuminating sequence that minimizes the spatial variability is proposed. The approach is tested for the classification of different color substances showing improvements up to 53% of the intra/inter cluster distance ratio measured in a PCA space, when compared to randomly chosen colors. © 2006 Elsevier B.V. All rights reserved.
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| 22. |
- Dini, Francesca, et al.
(författare)
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Combining porphyrins and pH indicators for analyte detection
- 2015
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Ingår i: Analytical and Bioanalytical Chemistry. - : Springer. - 1618-2642 .- 1618-2650. ; 407:14, s. 3975-3984
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Tidskriftsartikel (refereegranskat)abstract
- High sensitivity and cross-selectivity are mandatory properties for sensor arrays. Although metalloporphyrins and pH indicators are among the most common and appropriate choices for the preparation of optical sensor arrays, the sensitivity spectrum of these dyes is limited to those analytes able to induce an optical response. To extend the receptive field of optical sensors, we explore the design of composite materials, where the molecular interaction among the subunits enriches their sensing working mechanisms. We demonstrate that blends of single metalloporphyrins and pH indicators, tested with a transduction apparatus based on ubiquitous and easily available hardware, can be endowed with sensing properties wider than those of single constituents, enabling the recognition of a broad range of volatiles.
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| 23. |
- Enander, Karin, 1972-, et al.
(författare)
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A versatile polypeptide platform for integrated recognition and reporting : affinity arrays for protein-ligand interaction analysis
- 2004
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Ingår i: Chemistry - A European Journal. - : Wiley. - 0947-6539 .- 1521-3765. ; 10:10, s. 2375-2385
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Tidskriftsartikel (refereegranskat)abstract
- A molecular platform for protein detection and quantification is reported in which recognition has been integrated with direct monitoring of target-protein binding. The platform is based on a versatile 42-residue helix–loop–helix polypeptide that dimerizes to form four-helix bundles and allows site-selective modification with recognition and reporter elements on the side chains of individually addressable lysine residues. The well-characterized interaction between the model target-protein carbonic anhydrase and its inhibitor benzenesulfonamide was used for a proof-of-concept demonstration. An affinity array was designed where benzenesulfonamide derivatives with aliphatic or oligoglycine spacers and a fluorescent dansyl reporter group were introduced into the scaffold. The affinities of the array members for human carbonic anhydrase II (HCAII) were determined by titration with the target protein and were found to be highly affected by the properties of the spacers (dissociation constant Kd=0.02–3 μM). The affinity of HCAII for acetazolamide (Kd=4 nM) was determined in a competition experiment with one of the benzenesulfonamide array members to address the possibility of screening substance libraries for new target-protein binders. Also, successful affinity discrimination between different carbonic anhydrase isozymes highlighted the possibility of performing future isoform-expression profiling. Our platform is predicted to become a flexible tool for a variety of biosensor and protein-microarray applications within biochemistry, diagnostics and pharmaceutical chemistry.
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| 31. |
- Filippini, Daniel, 1968-, et al.
(författare)
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Chemical Sensing with Familiar Devices
- 2006
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Ingår i: Angewandte Chemie : : a journal of the Gesellschaft Deutscher Chemiker.. International edition. - : Wiley. ; 45:23, s. 3800-3803
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Tidskriftsartikel (refereegranskat)
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| 32. |
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| 33. |
- Filippini, Daniel, 1968-, et al.
(författare)
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Computer screen photo-assisted techniques
- 2005
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Ingår i: Encyclopedia of Sensors. - California, USA : American Scientific Publishers. - 9781588830562 - 158883056X ; , s. -8000
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Bokkapitel (övrigt vetenskapligt/konstnärligt)abstract
- The applications of sensors range from medical diagnostics to industrial manufacturing and to defense and national security applications. When an area spans such a large diversity of research, and where research from many different countries is also involved, a review of these developments becomes especially useful. Because it bridges science and technology the field also provides a desired interaction between researchers and research in technologically advanced and developing countries. The present series of volumes, "The Encyclopedia of Sensors" , the first of its kind, is intended to provide a timely compendium of the entire field. As such it can be expected to play a significant role in worldwide future progress and understanding."
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| 34. |
- Filippini, Daniel, 1968-, et al.
(författare)
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Computer screen photo-assisted techniques for global monitoring of environmental and sanitary parameters
- 2007
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Ingår i: Sensors and actuators. B, Chemical. - : Elsevier BV. - 0925-4005 .- 1873-3077. ; 121:1, s. 93-102
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Tidskriftsartikel (refereegranskat)abstract
- The computer screen photo-assisted technique (CSPT) is a measuring method that uses regular computer screens as controlled light sources and web cameras as imaging detectors for optical evaluations of sensing substances, which optical properties respond to target analytes. The fact, that CSPT uses a globally distributed and familiar infrastructure as instrumentation and that is able to serve multiple targets with the same setup, makes it an attractive candidate for home testing. This article reviews the CSPT concept and focuses in two recent studies that demonstrate thorough CSPT determinations using a handy and eventually disposable assay format, with the potential to be as ubiquitous as the CSPT platforms themselves. The implications of such combination of technologies is discussed, and its potential to support the global monitoring of environmental and sanitary parameters, using Internet geography browsers, is analyzed. © 2006 Elsevier B.V. All rights reserved.
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| 39. |
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| 41. |
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| 42. |
- Filippini, Daniel, 1968-, et al.
(författare)
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Effect of fingerprint conformation and spectral scaling on the performance of computer screen photo-assisted experiments
- 2006
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Ingår i: The Analyst. - : Royal Society of Chemistry (RSC). - 0003-2654 .- 1364-5528. ; 131:1, s. 118-125
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Tidskriftsartikel (refereegranskat)abstract
- The use of computer screens as controlled light sources and web cameras as image detectors (the so-called computer screen photo-assisted technique, CSPT) is an ubiquitous alternative for the evaluation of colorimetric quick tests at homes or in primary care units. The performance of CSPT for such evaluations depends on several factors, from which the most relevant are the composition of illuminating sequences and the conformation of CSPT substance signatures. In this work, with the aid of a CSPT model, the effect of the construction of the substance signatures on the classification performance of different representative substance sets is studied. The correlation of illuminating colors with such classification is investigated, allowing one to determine redundancy and limitations with respect to visible spectroscopy. The concept of spectral scaling is introduced and its properties compared with standard procedures. © The Royal Society of Chemistry 2006.
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| 43. |
- Filippini, Daniel, 1968-, et al.
(författare)
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ELISA test for anti-neutrophil cytoplasm antibodies detection evaluated by a computer screen photo-assisted technique
- 2005
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Ingår i: Biosensors & bioelectronics. - : Elsevier BV. - 0956-5663 .- 1873-4235. ; 21:2, s. 266-272
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Tidskriftsartikel (refereegranskat)abstract
- The computer screen photo-assisted technique (CSPT), a method for substance classification based on spectral fingerprinting, which involves just a computer screen and a web camera as measuring platform is used here for the evaluation of a prospective enzyme-linked immunosorbent assay (ELISA). A anti-neutrophil cytoplasm antibodies (ANCA-ELISA) test, typically used for diagnosing patients suffering from chronic inflammatory disorders in the skin, joints, blood vessels and other tissues is comparatively tested with a standard microplate reader and CSPT, yielding equivalent results at a fraction of the instrumental costs. The CSPT approach is discussed as a distributed measuring platform allowing decentralized measurements in routine applications, whereas keeping centralized information management due to its natural network embedded operation. © 2004 Elsevier B.V. All rights reserved.
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| 44. |
- Filippini, Daniel, 1968-, et al.
(författare)
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Fingerprinting of fluorescent substances for diagnostic purposes using computer screen illumination
- 2005
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Ingår i: Sensors and actuators. B, Chemical. - : Elsevier. - 0925-4005 .- 1873-3077. ; 106, s. 302-310
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Tidskriftsartikel (refereegranskat)abstract
- Spectral fingerprinting of representative fluorescent substances prospective of potential assays is demonstrated by using a computer screen photo-assisted technique (CSPT) that involves standard computer sets and web cameras as only measuring platform.The unfolding of the recorded information along the color channels of the web camera enables to generate distinctive substance features that retain key spectral information (spectral fingerprints), which are able to disentangle predominant emission from absorption features. Principal component analysis (PCA) of the CSPT measurements and of standard visible absorption/emission spectroscopy performs similarly, allowing substance classification with a ubiquitous and inexpensive resource suitable with home test evaluations.
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| 45. |
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| 46. |
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| 47. |
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| 48. |
- Filippini, Daniel, 1968-, et al.
(författare)
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Measurement strategy and instrumental performance of a computer screen photo-assisted technique for the evaluation of a multi-parameter colorimetric test strip
- 2006
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Ingår i: The Analyst. - : Royal Society of Chemistry (RSC). - 0003-2654 .- 1364-5528. ; 131:1, s. 111-117
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Tidskriftsartikel (refereegranskat)abstract
- A measuring strategy for the evaluation of a seven parameters colorimetric test using a computer screen photo-assisted technique (CSPT) is demonstrated. CSPT is a versatile approach aimed at point of care or home tests that uses regular computer sets and web cameras as the whole instrument. Issues such as the stability and the equivalency on different platforms of the determinations have been addressed in the present work. The method uses an embedded local reference simultaneously measured with the tests and solves the evaluation as a classification problem. The achieved performance tested along 580 classifications covering all the ranges of the assay, using synthetic samples, yielded 97.2% correct determinations compared with 89.7% for the case of colorimetric determinations. The errors were concentrated in only two parameters that show a significant correlation with a set of quality indices used to assess the performance of the classification. © The Royal Society of Chemistry 2006.
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| 49. |
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| 50. |
- Filippini, Daniel, 1968-, et al.
(författare)
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Natural nanosystems
- 2007
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Ingår i: International Symposium on Biomolecular Nanoscale Assemblies,2007.
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Konferensbidrag (refereegranskat)
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