2006
DOI: 10.1038/nbt1250
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Analyzing proteome topology and function by automated multidimensional fluorescence microscopy

Abstract: Temporal and spatial regulation of proteins contributes to function. We describe a multidimensional microscopic robot technology for high-throughput protein colocalization studies that runs cycles of fluorescence tagging, imaging and bleaching in situ. This technology combines three advances: a fluorescence technique capable of mapping hundreds of different proteins in one tissue section or cell sample; a method selecting the most prominent combinatorial molecular patterns by representing the data as binary ve… Show more

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Cited by 439 publications
(669 citation statements)
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“…9). Extensive literature exists on the development of multiplexed and more recently hyperplexed fluorescence methods for tissue analyses, including instrumentation, software, and bioinformatics tools that can be used in the development of diagnostic tests 9, 20, 21, 22, 23. The ability to measure many parameters in large numbers of tissue samples has propelled these assays into the realm of big data challenges for the future.…”
Section: Hyperplexed Fluorescence Measurements Of the Cellular And Sumentioning
confidence: 99%
See 1 more Smart Citation
“…9). Extensive literature exists on the development of multiplexed and more recently hyperplexed fluorescence methods for tissue analyses, including instrumentation, software, and bioinformatics tools that can be used in the development of diagnostic tests 9, 20, 21, 22, 23. The ability to measure many parameters in large numbers of tissue samples has propelled these assays into the realm of big data challenges for the future.…”
Section: Hyperplexed Fluorescence Measurements Of the Cellular And Sumentioning
confidence: 99%
“…Multiplex protein expression (4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37) …”
mentioning
confidence: 99%
“…Described previously, the multi-epitope-ligand cartography (MELC) 8,9 , is one of these bleaching techniques, that is capable to map the location of different proteins in one sample of cells or tissues using sequential rounds of fluorescent detection 10 . In each cycle, a couple of antibodies are added; phase contrast and fluorescence images then are acquired by a high-sensitivity "charged Coupled Device", a sensor used in digital cameras to record images; the sample is washed with phosphate buffer saline and bleaches at the excitation wavelengths; and post bleaching phase contrast and fluorescence images are acquired.…”
Section: Multi-epitope-ligand Cartographymentioning
confidence: 99%
“…This can be exploited to develop new diagnostic methods. Where single genetic markers are not sufficient to distinguish healthy tissue from a diseased phenotype, combinations of genes can enhance diagnostic strength [5]. Microscopy and comparative analysis of images is therefore becoming an important tool to study the spatio-temporal distribution of multiple gene products.…”
Section: Introductionmentioning
confidence: 99%