{"title":"在溶液中实现空间分辨单分子检测的步骤","authors":"H P Mathis, G Kalusche, B Wagner, J S McCaskill","doi":"10.1002/1361-6374(199709)5:3<116::AID-BIO5>3.0.CO;2-J","DOIUrl":null,"url":null,"abstract":"<p>Spatially resolved single molecule detection in solution is a prerequisite for molecule tracking and provides new opportunities in single molecule manipulation, for example in a sample flow. A detector with fast timing properties and high spatial resolution is required for these purposes. We introduce a concept for spatially resolved optical single molecule detection in an epi-illuminated microscope using a novel kind of detector and a new algorithm in configurable hardware for intelligent data processing. The analysis is performed on a parallel hardware interface and includes burst detection. It can be extended to <i>on-line</i> spatial and temporal data analysis. This detector will be used in combination with a molecular sorter where molecules are sorted in a microstructured flow device made of silicon. In a first step towards such sorting in these microstructures, we show that a reliable detection of single molecules in silicon microstructures is possible in zero-dimensional detection volumes. The noise structure of the data is analysed in terms of Poissonian statistics and it is shown that in the smallest structure used (depth 20 μm) a signal-to-noise ratio of 40 is achieved.</p>","PeriodicalId":100176,"journal":{"name":"Bioimaging","volume":"5 3","pages":"116-128"},"PeriodicalIF":0.0000,"publicationDate":"2001-05-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/1361-6374(199709)5:3<116::AID-BIO5>3.0.CO;2-J","citationCount":"0","resultStr":"{\"title\":\"Steps towards spatially resolved single molecule detection in solution\",\"authors\":\"H P Mathis, G Kalusche, B Wagner, J S McCaskill\",\"doi\":\"10.1002/1361-6374(199709)5:3<116::AID-BIO5>3.0.CO;2-J\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p>Spatially resolved single molecule detection in solution is a prerequisite for molecule tracking and provides new opportunities in single molecule manipulation, for example in a sample flow. A detector with fast timing properties and high spatial resolution is required for these purposes. We introduce a concept for spatially resolved optical single molecule detection in an epi-illuminated microscope using a novel kind of detector and a new algorithm in configurable hardware for intelligent data processing. The analysis is performed on a parallel hardware interface and includes burst detection. It can be extended to <i>on-line</i> spatial and temporal data analysis. This detector will be used in combination with a molecular sorter where molecules are sorted in a microstructured flow device made of silicon. In a first step towards such sorting in these microstructures, we show that a reliable detection of single molecules in silicon microstructures is possible in zero-dimensional detection volumes. The noise structure of the data is analysed in terms of Poissonian statistics and it is shown that in the smallest structure used (depth 20 μm) a signal-to-noise ratio of 40 is achieved.</p>\",\"PeriodicalId\":100176,\"journal\":{\"name\":\"Bioimaging\",\"volume\":\"5 3\",\"pages\":\"116-128\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2001-05-11\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://sci-hub-pdf.com/10.1002/1361-6374(199709)5:3<116::AID-BIO5>3.0.CO;2-J\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Bioimaging\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://onlinelibrary.wiley.com/doi/10.1002/1361-6374%28199709%295%3A3%3C116%3A%3AAID-BIO5%3E3.0.CO%3B2-J\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Bioimaging","FirstCategoryId":"1085","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/1361-6374%28199709%295%3A3%3C116%3A%3AAID-BIO5%3E3.0.CO%3B2-J","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
Steps towards spatially resolved single molecule detection in solution
Spatially resolved single molecule detection in solution is a prerequisite for molecule tracking and provides new opportunities in single molecule manipulation, for example in a sample flow. A detector with fast timing properties and high spatial resolution is required for these purposes. We introduce a concept for spatially resolved optical single molecule detection in an epi-illuminated microscope using a novel kind of detector and a new algorithm in configurable hardware for intelligent data processing. The analysis is performed on a parallel hardware interface and includes burst detection. It can be extended to on-line spatial and temporal data analysis. This detector will be used in combination with a molecular sorter where molecules are sorted in a microstructured flow device made of silicon. In a first step towards such sorting in these microstructures, we show that a reliable detection of single molecules in silicon microstructures is possible in zero-dimensional detection volumes. The noise structure of the data is analysed in terms of Poissonian statistics and it is shown that in the smallest structure used (depth 20 μm) a signal-to-noise ratio of 40 is achieved.