Finite conjugate spherical aberration compensation in high numerical-aperture optical disc readout

Stallinga, Sjoerd

doi:10.1364/ao.44.007307

Cited by 16 publications

(16 citation statements)

References 11 publications

(13 reference statements)

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“…The experimental setup is illustrated in The axial magnification of the imaging path is M ax = χM 2 /n = 352, where a sample refractive index n = 1.5 is assumed and χ = 1.07 is a non-paraxial correction factor [32,33]. The specimen (slide) is scanned using two stages: a PI M-505 low profile translation stage for positioning of the slide in the field direction and a Newport XM1000 ultra precision linear motor stage for the continuous scanning motion of the sample.…”

Section: Methodsmentioning

confidence: 99%

Multi-line fluorescence scanning microscope for multi-focal imaging with unlimited field of view

Graaff¹,

Leenders²,

Boyaval³

et al. 2019

Biomed. Opt. Express

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Confocal scanning microscopy is the de facto standard modality for fluorescence imaging. Point scanning, however, leads to a limited throughput and makes the technique unsuitable for fast multi-focal scanning over large areas. We propose an architecture for multifocal fluorescence imaging that is scalable to large area imaging. The design is based on the concept of line scanning with continuous 'push broom' scanning. Instead of a line sensor, we use an area sensor that is tilted with respect to the optical axis to acquire image data from multiple depths inside the sample simultaneously. A multi-line illumination where the lines span a plane conjugate to the tilted sensor is created by means of a diffractive optics design, implemented on a spatial light modulator. In particular, we describe a design that uses higher order astigmatism to generate focal lines of substantially constant peak intensity along the lines. The proposed method is suitable for fast 3D image acquisition with unlimited field of view, it requires no moving components except for the sample scanning stage, and provides intrinsic alignment of the simultaneously scanned focal slices. As proof of concept, we have scanned 9 focal slices simultaneously over an area of 36 mm 2 at 0.29 µm pixel size in object space. The projected ultimate throughput that can be realized with the proposed architecture is in excess of 100 Mpixel/s.

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Section: Methodsmentioning

confidence: 99%

Multi-line fluorescence scanning microscope for multi-focal imaging with unlimited field of view

Graaff¹,

Leenders²,

Boyaval³

et al. 2019

Biomed. Opt. Express

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“…It follows that in this case we will suffer from spherical aberration. The sensitivity to spherical aberration can be analyzed along the lines of [43]. In that analysis, it is assumed that the objective lens and tube lens form an aplanatic telecentric imaging system.…”

Section: B Finite Conjugate Imaging and Spherical Aberrationmentioning

confidence: 99%

Computational imaging modalities for multi-focal whole-slide imaging systems

et al. 2020

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Whole-slide imaging systems can generate full-color image data of tissue slides efficiently, which are needed for digital pathology applications. This paper focuses on a scanner architecture that is based on a multi-line image sensor that is tilted with respect to the optical axis, such that every line of the sensor scans the tissue slide at a different focus level. This scanner platform is designed for imaging with continuous autofocus and inherent color registration at a throughput of the order of 400 MPx/s. Here, single-scan multi-focal whole-slide imaging, enabled by this platform, is explored. In particular, two computational imaging modalities based on multi-focal image data are studied. First, 3D imaging of thick absorption stained slides (∼60 µm) is demonstrated in combination with deconvolution to ameliorate the inherently weak contrast in thick-tissue imaging. Second, quantitative phase tomography is demonstrated on unstained tissue slides and on fluorescently stained slides, revealing morphological features complementary to features made visible with conventional absorption or fluorescence stains. For both computational approaches simplified algorithms are proposed, targeted for straightforward parallel processing implementation at ∼GPx/s throughputs.

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“…Shifted intensity peaks are useful for sample illumination but suffer from spherical aberration as described. 5 There it is shown that finite conjugate illumination has aberration W, given by…”

Section: Finite Conjugate Illuminationmentioning

confidence: 99%

“…These observations demonstrate profile adaptability by use of the LFL. Finite conjugate illumination was used to compensate for aberrations introduced by specimen refractive index inhomogeneities 5 and could likewise be used on biological samples to correct for sample inhomogeneities affecting excitation focus. The objective correction collar nominally corrects for temperature and coverslip thickness variability.…”

Section: Introductionmentioning

confidence: 99%

“…The exciting light intensity profile is strictly proportional to fluorescence detected, confirming the bimodal axial intensity distribution for the two-beam configuration. The objective is not translating axially, hence no emission profile or emission aberration is involved; however, there is aberration in the exciting profile as discussed previously 5 that I fully account for in simulation. The LFL translates the exciting light axial profile such that the peak intensity and emission focal plane are brought into coincidence.…”

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Evanescent field shapes excitation profile under axial epi-illumination

Burghardt

2012

J. Biomed. Opt.

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Abstract. Axial epi-illuminating light transmitting a >1.3-numerical-aperture microscope objective creates an excitation volume at focus with size and shape dictated by diffraction and due to refraction by the objective and by the coverslip interface separating a specimen in aqueous buffer from the oil immersion objective. The evanescent field on the coverslip aqueous side affects primarily the excitation volume axial dimension as the specimen in focus approaches the interface to within a few hundred nanometers. Following excitation, an excited stationary dipole moment emits fluorescence in a spatially varying pattern collected over the large objective aperture. Collected light propagates in parallel rays toward the tube lens that forms a real three-dimensional image that is decoded to identify dipole orientation. An integral representation of the excitation and emitted fields for infinity-corrected optics-including effects of finite conjugate illumination, fluorescence emission near an interface, emitter dipole orientation, spherical aberration, light transmission through a dichroic filter, and for real microscopic specifications-accurately models observed field intensities including the substantial excitation from the evanescent field. The goal is to develop and verify the practical depiction of excitation and emission in a real microscope for quantitative interpretation of the 3-D emission pattern.

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Finite conjugate spherical aberration compensation in high numerical-aperture optical disc readout

Cited by 16 publications

References 11 publications

Multi-line fluorescence scanning microscope for multi-focal imaging with unlimited field of view

Multi-line fluorescence scanning microscope for multi-focal imaging with unlimited field of view

Computational imaging modalities for multi-focal whole-slide imaging systems

Evanescent field shapes excitation profile under axial epi-illumination

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