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E-book Application of Diffractive Lens Arrays in Confocal Microscopy

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Penilaian

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Since its first invention by Minsky in the 1950s [Min61], confocal microscopyhas become one of the most important advances of light microscopy in the pastdecades [Wil09]. Confocal microscopes have the advantages over conventionalwide-field microscopes in resolution and contrast, making them widely usedin biomedical imaging and industrial metrology.The setup of a typical reflected-light scanning confocal microscope is shownin Figure 1.1 (a). A point light source, usually a laser, is collimated and focusedto a spot by an objective. The spot illuminates a tiny part of the sample. Then,it is imaged by the objective onto the detector. A small pinhole is used to block the out-of-focus light scattered by the other parts of the sample. In this way,the sample is scanned by the spot and a sharper image can be reconstructed.Confocal microscopes can image the sample with higher resolution than con-ventional wide-field microscopes in both the lateral and the axial directions.The pinhole can be replaced by a pinhole array to increase the measurementspeed by multi-spot scanning. For example, a typical Yokogawa spinning-diskconfocal microscope is shown in Figure 1.1 (b) [Fav92, Ich96]. A Nipkow disk[Nip84] rotates along the axis and the sample is scanned by multiple spotswhich are produced by the pinholes on the disk. Comparing to the single-spotscanning, it can achieve a higher measurement speed. where????is the refractive index of the medium and????is the half angle of thelight cone focused by the objective. According to the well-known resolutionlimit, which was first described by Abbe in 1873 [Abb73] and later refinedby Lord Rayleigh in 1879 [FRS79], the smallest resolvable distance betweentwo points which are imaged by a microscope is inversely proportional tothe NA of the objective where?????is the minimum resolvable distance and????is the wavelength. Sucha relation is also widely known as the Rayleigh criterion.Subsequently, high-NA objectives are generally required in order to image thesample with high resolution, while they usually also have large magnifications,which lead to small fields of view (FOVs). The relation between the resolutionand the FOV of an optical system can be described by the space-bandwidthproduct (SBP), which is, in the two-dimensional case, defined as by the detector noise and sensitivity [Ore14] in certain applications such asfluorescence microscopy with low light intensities.Conversely, multi-spot scanning confocal microscopes utilize parallel scanningwith camera sensors which have higher quantum efficiencies to realize higherframe rates [Too06]. Traditionally, the projection of multiple spots is achievedby pinhole arrays or beam splitting in front of the objectives. The most commontype is the spinning-disk setup which was first introduced by Petran et al.[Pet68] and later modified by other researchers [Fav92, Ich96, Ino02, Tan02].Besides, a number of papers [Pac17, Ada18, Bes18] and patents [Lun08, Abr16,Sch20, Mat21] have used various kinds of diffractive optical elements (DOEs),e.g., Dammann gratings [Jah89], to split the illumination beam into multiplebeams with different incident angles. Then, the beams are also scanned by thetraditional galvanometer mirror scanners as the CLSMs. Nevertheless, suchmulti-spot confocal microscope configurations still rely on the objectives toproject spots. Consequently, they remain to suffer from the trade-offs betweenFOVs and NAs, and thus they are limited in SBPs.

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Ketersediaan

Informasi Detail

Judul Seri

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No. Panggil

502.82 ZHE a

Penerbit

Berlin : KIT Scientific Publishing., 2022

Deskripsi Fisik

178 hlm

Bahasa

English

ISBN/ISSN

9783731511885

Klasifikasi

502.82

Tipe Isi

text

Tipe Media

computer

Tipe Pembawa

online resource

Edisi

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Subjek

Microscopes

Info Detail Spesifik

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Pernyataan Tanggungjawab

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Versi lain/terkait

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