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    188宝金博页面版: Separation in the Time Domain

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    内容提示： 9Separation in the Time DomainThus far we did not pay too much attention to the f l uorescence lifetime (Sect. 2.1).As the lifetime of the excited state is in the range of a few nanoseconds, andtherefore not easy to capture by measurement devices, for a long time it was not astandard parameter that would be used to characterize f l uorescence phenomena.This has changed in the last 20 years, and measurements based on f l uorescencelifetime are found more frequently in the literature. In this chapter we disc...

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    9Separation in the Time DomainThus far we did not pay too much attention to the f l uorescence lifetime (Sect. 2.1).As the lifetime of the excited state is in the range of a few nanoseconds, andtherefore not easy to capture by measurement devices, for a long time it was not astandard parameter that would be used to characterize f l uorescence phenomena.This has changed in the last 20 years, and measurements based on f l uorescencelifetime are found more frequently in the literature. In this chapter we discusssimple lifetime measurements and how we can benef i t from these relationships ineveryday microscopy.9.1 Sensors Sense PhotonsApart from occasionally used “sensor bars” in some spectral instruments, confocalmicroscopy is performed with single (point) sensors. The spot of light is movedpoint by point over the microscopic f i eld, and the measured electrical signal issynchronously stored in an electronic device as a two-dimensional image. Mostoften, photomultiplier tubes (PMT) are used as a sensor (see also Sect. 3.3.1).Occasionally, especially for single molecule applications, avalanche photodiodesare employed (APD). Lately, hybrid detectors (HPD, HyD) have been utilized withgreat success. The various types of sensors differ in wavelength-dependent quantumeff i ciency, useful dynamic range, and temporal resolution. The latter is of particularinterest in lifetime measurements, and the three concepts are discussed in respectthereof.© Springer International Publishing AG 2017R.T. Borlinghaus, The White Confocal,DOI 10.1007/978-3-319-55562-1_9107

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