1、 g49g50g3g38g50g51g60g44g49g42g3g58g44g55g43g50g56g55g3g37g54g44g3g51g40g53g48g44g54g54g44g50g49g3g40g59g38g40g51g55g3g36g54g3g51g40g53g48g44g55g55g40g39g3g37g60g3g38g50g51g60g53g44g42g43g55g3g47g36g58microscopy Part 2: Advanced techniques in light microscopyICS 01.040.37; 37.020Optics and optical i
2、nstruments Vocabulary for BRITISH STANDARDBS ISO 10934-2:2007BS ISO 10934-2:2007This British Standard was published under the authority of the Standards Policy and Strategy Committee on 30 March 2007 BSI 2007ISBN 978 0 580 50409 9Amendments issued since publicationAmd. No. Date Commentscontract. Use
3、rs are responsible for its correct application.Compliance with a British Standard cannot confer immunity from legal obligations. National forewordThis British Standard was published by BSI. It is the UK implementation of ISO 10934-2:2007.The UK participation in its preparation was entrusted by Techn
4、ical Committee CPW/172, Optics and photonics, to Subcommittee CPW/172/5, Microscopes.A list of organizations represented on CPW/172/5 can be obtained on request to its secretary.This publication does not purport to include all the necessary provisions of a Reference numberISO 10934-2:2007(E)INTERNAT
5、IONAL STANDARD ISO10934-2First edition2007-03-01Optics and optical instruments Vocabulary for microscopy Part 2: Advanced techniques in light microscopyOptique et instruments doptique Vocabulaire relatif la microscopie Partie 2: Techniques avances en microscopie optique BS ISO 10934-2:2007ii iiiFore
6、word ISO (the International Organization for Standardization) is a worldwide federation of national standards bodies (ISO member bodies). The work of preparing International Standards is normally carried out through ISO technical committees. Each member body interested in a subject for which a techn
7、ical committee has been established has the right to be represented on that committee. International organizations, governmental and non-governmental, in liaison with ISO, also take part in the work. ISO collaborates closely with the International Electrotechnical Commission (IEC) on all matters of
8、electrotechnical standardization. International Standards are drafted in accordance with the rules given in the ISO/IEC Directives, Part 2. The main task of technical committees is to prepare International Standards. Draft International Standards adopted by the technical committees are circulated to
9、 the member bodies for voting. Publication as an International Standard requires approval by at least 75 % of the member bodies casting a vote. Attention is drawn to the possibility that some of the elements of this document may be the subject of patent rights. ISO shall not be held responsible for
10、identifying any or all such patent rights. ISO 10934-2 was prepared by Technical Committee ISO/TC 172, Optics and photonics, Subcommittee SC 5, Microscopes and endoscopes. ISO 10934 consists of the following parts, under the general title Optics and optical instruments Vocabulary for microscopy: Par
11、t 1: Light microscopy Part 2: Advanced techniques in light microscopy BS ISO 10934-2:2007blank1Optics and optical instruments Vocabulary for microscopy Part 2: Advanced techniques in light microscopy 1 Scope This part of ISO 10934 specifies terms and definitions to be used in the field of advanced l
12、ight microscopy. 2 Terms and definitions NOTE Terms shown in bold within a definition or a note are defined elsewhere within this part of ISO 10934. 2.1 acousto-optical modulator electronically-tunable device used to control the direction and/or intensity of a laser by an acoustically-induced diffra
13、ction grating in a crystal 2.2 acousto-optical tunable filter AOTF electronically-tunable filter for selection of wavelengths by an acoustically-induced diffraction grating in a crystal 2.3 aliasing phenomenon caused by sampling at too low a frequency (i.e. lower than the Nyquist frequency) resultin
14、g in the loss of information and/or the creation of spurious information 2.4 auto-focus method of bringing an object automatically into focus, controlled by an imaging software algorithm and/or a hardware device that detects the object position 2.5 axial resolution resolution in the direction of the
15、 optical axis 2.6 background subtraction removal of that part of the signal that is present in the absence of the object, to reveal underlying image information 2.7 binning mode of operation of an image sensor where the charge of adjacent pixels is accumulated and is read out as a single value BS IS
16、O 10934-2:20072 2.8 confocal microscopy state in which, ideally, a point in the object field is illuminated by a diffraction-limited spot of light, and light emanating from this point is focused upon and detected from an area smaller than the central area of the diffraction disc situated in the corr
17、esponding position in a subsequent field plane 2.9 channel particular signal path containing one type of image information 2.10 co-localization overlay of images with coincidence of pixels corresponding to the same object points 2.11 confocal microscope microscope in which, ideally, a point in the o
18、bject plane is illuminated by a diffraction-limited spot of light, and light emanating from this point is focused upon and detected from an area smaller than the central area of the diffraction disc situated in the corresponding position in a subsequent field plane NOTE 1 An image of an extended are
19、a is formed either by scanning the object, or by scanning the illuminated and detected spots simultaneously. NOTE 2 The confocal principle leads to improved axial resolution by suppression of light from out-of-focus planes. 2.11.1 laser-scanning confocal microscope confocal microscope (2.11) in whic
20、h the light source is a laser 2.11.2 multiple-beam confocal microscope confocal microscope (2.11) using more than one illuminated and detected spot simultaneously 2.11.3 Nipkow disc confocal microscope confocal microscope (2.11) in which the scanning of the illuminated and detected spots is performe
21、d using a Nipkow disc (2.11.3.1) 2.11.3.1 Nipkow disc opaque disc with many ideally identical small holes arranged in Archimedean spirals 2.11.3.2 Tandem-scanning confocal microscope Nipkow disc confocal microscope (2.11.3) in which the illuminating light and the detected light pass through separate
22、 holes 2.11.4 spectral confocal microscope confocal microscope (2.11) in which a spectrum is recorded corresponding to spatial positions in an object 2.11.5 theta confocal microscope confocal microscope (2.11) in which two objectives positioned at an angle, , with respect to one another, and with fo
23、cal points coincident in the object, are used for excitation and collection respectively 2.11.6 white-light confocal microscope confocal microscope (2.11) using an illumination source and a detector operating throughout the visible spectrum BS ISO 10934-2:200732.11.7 confocal point spread function p
24、roduct of the point spread functions (2.35) of the illuminating and detecting optical systems in a confocal microscope (2.11) 2.11.8 confocal volume effective volume around each point in the object which gives rise to the image in a confocal microscope (2.11) 2.11.9 4 Pi confocal microscope confocal
25、 microscope (2.11) in which two opposing objective lenses with focal points coincident in the object are used to produce interference in the focal region from which an image signal is derived, and with further processing produces an image with enhanced axial resolution (2.5) 2.12 deconvolution micro
26、scopy mathematical method for reducing blur, performed either in the spatial domain, or in the frequency domain by inverse filtering techniques NOTE If the deconvolution is based solely on theoretical as opposed to measured values it is known as blind deconvolution. 2.13 digitally enhanced contrast
27、contrast enhanced by manipulation of the intensity and/or colour values in a digital image (2.14) 2.14 digital image image in which the information is in the form of binary or other machine code 2.15 electronic image image in which the information is in the form of electrical signals 2.16 extended d
28、epth of field microscopy microscopy in which the point spread function (2.35) is modified in a known fashion such that it becomes substantially invariant over an extended focal range, and by further processing results in an image with extended depth of field 2.17 extended focus image image processin
29、g two-dimensional image derived by summing the pixel intensity values in a projection through an image stack (2.28.1) 2.18 fluorescence correlation microscopy microscopy in which time-dependant intensity fluctuations occurring within a confocal volume (2.11.8) are used to calculate the mobility of f
30、luorescent molecules 2.19 fluorescence in-situ hybridisation microscopy FISH microscopy in which chromosomes or specific positions within chromosomes can be fluorescently labelled by in-situ hybridisation BS ISO 10934-2:20074 2.20 fluorescence life-time imaging FLIM imaging method based on discrimin
31、ating characteristic fluorescence decay rates 2.21 fluorescence recovery after photobleaching FRAP technique in which a region in the object is irradiated to deplete its fluorescence, the subsequent recovery of fluorescence in the irradiated region being measured 2.22 fluorescence resonance energy t
32、ransfer Frster resonance energy transfer FRET non-radiative transfer of energy between two fluorophores in close proximity 2.23 frame averaging averaging the pixel values from sequential images recorded under identical conditions NOTE Used to increase signal-to-noise ratio. 2.24 image intensifier de
33、vice which increases the dynamic range of a signal to match the range of the detector 2.25 linear array sensor detector in the form of a line of sensitive elements 2.26 maximum intensity image image processing two-dimensional image derived from the maximum pixel intensity values in a projection thro
34、ugh an image stack (2.28.1) 2.27 microchannel plate device positioned in front of a detector array to multiply incoming photon flux by secondary emission 2.28 multidimensional image data set image data generated by recording data from a sample using several parameters, e.g., three-space dimensions,
35、wavelength, time, polarization 2.28.1 image stack multidimensional image data set (2.28) acquired from a three-dimensional region of an object 2.28.2 focus series Z stack image stack (2.28.1) acquired at different focal positions 2.29 multi-mode fibre optical fibre that can sustain more than one tra
36、nsverse electromagnetic mode BS ISO 10934-2:200752.30 multi-photon fluorescence fluorescence excitation by the simultaneous absorption of multiple coherent photons 2.30.1 multi-photon fluorescence microscopy microscopy in which the image is formed by multi-photon fluorescence (2.30) NOTE Since suffi
37、cient excitation intensity is achieved only in a limited focal volume, multi-photon fluorescence results in optical sectioning without the need for a confocal pinhole. It also permits excitation by longer wavelengths. 2.30.1.1 two-photon fluorescence multi-photon fluorescence (2.30) excited by pairs
38、 of coherent photons 2.31 optical section image from a thin region whose thickness within a thick object is defined by the axial resolution (2.5) of the optical system 2.32 photobleaching destruction of fluorescing properties of molecules by light, resulting in reduced fluorescence of the sample 2.3
39、3 pinhole confocal microscopy diaphragm situated in a plane conjugate with the object, which restricts the area in the object plane that is illuminated and/or from which light is collected 2.34 point detection detection of light collected from a restricted point-like area of an image 2.35 point spre
40、ad function lens system mathematical expression of the distribution of the light amplitude or intensity in the image of a point source 2.36 Raman microscopy microscopy utilizing Raman scattering as the source of image information 2.37 ratio imaging forming an image in which the pixel values are obta
41、ined by dividing the corresponding pixel values of two images 2.38 real time imaging displaying or analysing images at the same rate as that at which they are collected NOTE This rate is normally also commensurate with the dynamics of the processes to be observed within the specimen and perceived to
42、 be continuous by the eye. 2.39 region of interest ROI parts of an image to which discrete observations are applied BS ISO 10934-2:20076 2.40 scan rate number of scan cycles completed per unit time 2.41 scanned field dimensions of the scanned area in object space 2.42 scanning sequential illuminatio
43、n of or detection from regions in an object NOTE Scanning may be accomplished by moving the object, illuminating beam(s), objective or detector(s). 2.42.1 descanning process by which an imaging beam retraces the path of the illuminating beam through the scanning (2.42) mechanism to produce a station
44、ary beam 2.42.2 line scanning scanning (2.42) along a single line 2.42.3 non-descanned detection NDD method of obtaining an image signal in a scanning microscope without descanning (2.42.1) NOTE Widely used in multiphoton fluorescence microscopy. 2.42.4 point scanning scanning (2.42) an area using a
45、 spot of light 2.42.5 raster scanning scanning (2.42) an area by a pattern of lines 2.42.6 slit scanning scanning (2.42) an area with a bar of light 2.42.7 stage scanning scanning (2.42) performed by moving the stage and hence the object 2.43 scanning microscope microscope in which the image is form
46、ed by scanning (2.42) 2.43.1 disc scanning microscope scanning microscope (2.43) in which scanning is achieved by means of a perforated disc rotated in the illumination and/or observation paths 2.43.2 laser-scanning microscope scanning microscope (2.43) in which the object is scanned by a laser beam
47、 BS ISO 10934-2:200772.43.3 scanning near-field microscope scanning microscope (2.43) in which a small light-emitting probe is scanned across the object (or vice versa) at a close distance NOTE When the probe diameter is smaller than the Airy disc, super-resolution (2.49) may be achieved. 2.44 secon
48、d harmonic generation microscopy microscopy in which the second harmonic of the excitation light is used to provide additional image information 2.45 shading correction method for adjusting intensity levels in an image, caused by non-uniformities in the illuminating or detecting systems 2.46 single-
49、mode fibre optical fibre that can sustain only a single transverse electromagnetic mode 2.47 spectral imaging microscopy microscopy in which a spectrum is recorded corresponding to spatial positions in an object 2.48 structured illumination microscope microscope in which the object is illuminated by a spatially varying pattern so as to produce a composite image from which images with enhanced lateral and axial resolution may be produced 2.48.1 grating image microscope structured illumination microscope (2.48) in which
