A common aberration with water‐immersion objective lenses

Arimoto, Rieko; Murray, John M.

doi:10.1111/j.0022-2720.2004.01383.x

Cited by 44 publications

(36 citation statements)

References 6 publications

(6 reference statements)

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“…As mentioned before, any difference in the coverslip thickness with respect to its expected and designed thickness, produces defocus and spherical aberration. Another coverslip related aberration has been discussed in [44], which mostly affects the water immersion microscope objectives. The tilt of the coverslip generates an asymmetry of the PSF, which is particularly detrimental for 3D deconvolution.…”

Section: Sample Holder Aberrationsmentioning

confidence: 99%

Light sheet adaptive optics microscope for 3D live imaging

Bourgenot

Taylor

Saunter

et al. 2013

Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XX

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The full-text may be used and/or reproduced, and given to third parties in any format or medium, without prior permission or charge, for personal research or study, educational, or not-for-prot purposes provided that:• a full bibliographic reference is made to the original source• a link is made to the metadata record in Durham E-Theses• the full-text is not changed in any wayThe full-text must not be sold in any format or medium without the formal permission of the copyright holders. DeclarationThe work in this thesis is based on research carried out at

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Section: Sample Holder Aberrationsmentioning

confidence: 99%

Light sheet adaptive optics microscope for 3D live imaging

Bourgenot

Taylor

Saunter

et al. 2013

Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XX

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“…16 Aberrations in biospecimens and their effects have been experimentally investigated by several authors. [17][18][19] Further, wavefront aberrations can be described using Zernike polynomials.…”

Section: Introductionmentioning

confidence: 99%

Transmissive liquid-crystal device for correcting primary coma aberration and astigmatism in biospecimen in two-photon excitation laser scanning microscopy

et al. 2016

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Nemoto, "Transmissive liquid-crystal device for correcting primary coma aberration and astigmatism in biospecimen in two-photon excitation laser scanning microscopy," J. Abstract. All aberrations produced inside a biospecimen can degrade the quality of a three-dimensional image in two-photon excitation laser scanning microscopy. Previously, we developed a transmissive liquid-crystal device to correct spherical aberrations that improved the image quality of a fixed-mouse-brain slice treated with an optical clearing reagent. In this study, we developed a transmissive device that corrects primary coma aberration and astigmatism. The motivation for this study is that asymmetric aberration can be induced by the shape of a biospecimen and/or by a complicated refractive-index distribution in a sample; this can considerably degrade optical performance even near the sample surface. The device's performance was evaluated by observing fluorescence beads. The device was inserted between the objective lens and microscope revolver and succeeded in improving the spatial resolution and fluorescence signal of a bead image that was originally degraded by asymmetric aberration. Finally, we implemented the device for observing a fixed whole mouse brain with a sloping surface shape and complicated internal refractive-index distribution. The correction with the device improved the spatial resolution and increased the fluorescence signal by ∼2.4×. The device can provide a simple approach to acquiring higher-quality images of biospecimens.

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“…Despite the slanted scan being capable of simultaneously scanning multiple depths of the specimen, the slide slant induces optical aberrations resembling that of a lens coma. The main artefacts include movement of the object blur at different depths (bending of the PSF) and the reduction of the image contrast compared with images acquired in the conventional flat orientation (Arimoto and Murray 2004;Bamford and Mayer 2009) as shown in Fig. 2.5.…”

Section: Slanted Scanmentioning

confidence: 99%

“…2.5 have been reported in fluorescence microscopy when using a water-immersion objective when the slide is slanted unintentionally (Arimoto and Murray 2004). Interestingly, the oil-immersion objective lens does not suffer from this type of aberration (Arimoto and Murray 2004) as the refractive index of oil is closely matched to that of the coverslip.…”

Section: Introductionmentioning

confidence: 96%

“…However, the slanted slide induces optical aberrations that resemble lens coma. The main artefacts include movement of objects in the image between different focal planes (due to bending of the optical point spread function) and a reduction in both image resolution and contrast compared to images acquired from the conventional flat scan (Arimoto and Murray 2004;Bamford and Mayer 2009). An example of this aberration is illustrated in Chapter 2 Fig.…”

Section: Introductionmentioning

confidence: 99%

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Methods for rapid and high quality acquisition of whole slide images

Fan¹

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In the past two decades, whole slide imaging (WSI) has been increasingly used in clinical pathology, research and education thanks to advances in technology. However, existing digital slide scanners for WSI are incapable of efficiently scanning cytology specimens in "glass-faithful quality". In particular, cervical cytology specimens used for cervical cancer screening need to be imaged at multiple focal planes and at high spatial resolution. So, the aim of this thesis is to develop novel methods for improving the scan efficiency of cytology specimens. Preliminary investigations narrowed the research into two directions: to optimise the scan parameters (e.g. the scan map, the focus profile map and the number of focal planes to be scanned) for scanning cytology specimens, and to investigate the fast slanted (specimen) scan.Creating an accurate scan map followed by estimating the focus profile map of the specimen can significantly reduce the scan time, but previous methods are not robust to slide artefacts such as ink marker and dusts. The first research aspect of this thesis has focused on the development of a robust method that not only creates an accurate scan map but also provides a ranked list of focus candidates required for estimating the focus map. Specifically, tiles representing field-of-views to be scanned at high resolution are located on the low resolution images and evaluated with regards to whether they contain foreground objects and how good they are as focus candidates. Four metrics were proposed for the tile evaluation: threshold index (TI) that measures the image intensity; normalised auto-correlation index (NACI) that measures the spatial image similarity; auto-phase correlation index (APCI) that measures the image phase diversity; and entropy index (EI) that measures the predictability of image intensities. The experimental results indicated that while NACI, APCI and EI are effective for specimen delineation, only APCI is capable of selecting superior focus candidates and ignoring artefacts.Knowing the three-dimensional distribution of the specimen can help to determine the minimal number of focal planes to be acquired, and hence the minimal scan time. A method has been developed to evaluate the thickness of all cell and cell clumps in cervical cytology specimens incorporating an exhaustive high-resolution scan, an over-complete wavelet transform across multi-focal planes and a cell clump segmentation method. The accuracy and usefulness of the method was evaluated and demonstrated by quantitative analysis of ten Pap stained Thin-prep cervical slides. In particular, it was found that creating a focus profile map was able to reduce the number of scanning focal planes from 25.4 to 21.4 on average. It was also shown that the distribution of cells was skewed towards the cover-slip (top of the slide), so by considering the thickness of the specimen, an improved focal map i can be produced to further reduce the required number of 1 micron spaced focal planes to 18.6.The last research aspect of...

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A common aberration with water‐immersion objective lenses

Cited by 44 publications

References 6 publications

Light sheet adaptive optics microscope for 3D live imaging

Light sheet adaptive optics microscope for 3D live imaging

Transmissive liquid-crystal device for correcting primary coma aberration and astigmatism in biospecimen in two-photon excitation laser scanning microscopy

Methods for rapid and high quality acquisition of whole slide images

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