Accuracy and perceptions of virtual microscopy compared with glass slide microscopy in cervical cytology

Evered, Andrew; Dudding, N.

doi:10.1111/j.1365-2303.2010.00758.x

Cited by 86 publications

(117 citation statements)

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“…A stack of 41 images (20 above and below the BSFP) were acquired at interval of 1µm for every non-empty FOV. The 41µm sampled depth range range was believed to be larger than the reported thickness of the specimens (15-20µm) [8]. To exclude any outlier FOVs, the segmentation results and depths maps were manually examined for quality, especially where the specimen was thick.…”

Section: Specimen Preparation and Imagingmentioning

confidence: 99%

“…In addition, important diagnostic cells are often found within these cell clumps, such as secretory cells from the endocervic and low-grade squamous intraepithelial lesions [15]. Therefore, it is no surprise to see that the acquisition of multiple focal planes achieves better diagnostic performances than acquiring only one or a few focal planes [26,8]. Additionally, interpreting cellular objects requires the microscope to operate at high magnification (40x), preferably at resolutions close to the diffraction limit (∼0.2µm/pixel), where the depth-of-field is reduced to around one micron [5].…”

Section: Introductionmentioning

confidence: 99%

“…For example, it was first shown that digital cervical cytology slides with seven focal planes achieved higher diagnostic accuracy than those of a single focal plane (both at 40X), but both were outperformed by conventional glass slides [26]. Another study reported that cervical specimens scanned with 21 focal planes at a 1.5µm interval achieved better diagnostic accuracy than those scanned with 5 focal planes at 1µm interval [8]. A more recent work argues that scanning only three focal planes (with 1µm interval) is able to achieve diagnostic performances close to that of a conventional glass slide [6].…”

Section: Introductionmentioning

confidence: 99%

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A method for quantitative analysis of clump thickness in cervical cytology slides

Fan

Bradley

2016

Micron

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A c c e p t e d M a n u s c r i p tWe developed an algorithm to estimate the thickness of cervical cytology specimens.The proposed algorithm reached an accuracy of 1 micron at 90% of times.The algorithm was used to quantitatively analysis of ten normal Thin-prep cervical cytology slides.It was found that the distribution of cells is skewed towards the cover-slip (top of the slide).It was also proved that considering the thickness of focal points produced focus maps in superior qualities compared to conventional ones. Highlights (for review)Page 2 of 28 A c c e p t e d M a n u s c r i p t AbstractKnowledge of the spatial distribution and thickness of cytology specimens is critical to the development of digital slide acquisition techniques that minimise both scan times and image file size. In this paper, we evaluate a novel method to achieve this goal utilising an exhaustive high-resolution scan, an over-complete wavelet transform across multi-focal planes and a clump segmentation of all cellular material on the slide. The method is demonstrated with a quantitative analysis of ten normal, but difficult to scan Pap stained, Thin-prep, cervical cytology slides. We show that with this method the top and bottom of the specimen can be estimated to an accuracy of 1 micron in 88% and 97% of the fields of view respectively. Overall, cellar material can be over 30 microns thick and the distribution of cells is skewed towards the cover-slip (top of the slide). However, the median clump thickness is 10 microns and only 31% of clumps contain more than three nuclei. Therefore, by finding a focal map of the specimen the number of 1 micron spaced focal planes that are required to be scanned to acquire 95% of the in-focus material can be reduced from 25.4 to 21.4 on average. In addition, we show that by considering the thickness of the specimen, an improved focal map can be produced which further reduces the required number of 1 micron spaced focal planes to 18.6. This has the potential to reduce scan times and raw image data by over 25%.

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Section: Specimen Preparation and Imagingmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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A method for quantitative analysis of clump thickness in cervical cytology slides

Fan

Bradley

2016

Micron

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“…It is generally believed that the thickness of the LBC based cervical cytology specimens used for cervical cytology screening is around 20 microns, so around 20 focal planes are acquired in some studies (Bradley and Bamford 2004;Evered and Dudding 2011). Although some studies had argued that fewer number of focal planes could be sufficient for diagnosis, the exact number of focal planes required is generally decided empirically.…”

Section: Specimen Localisationmentioning

confidence: 99%

“…In addition, important diagnostic cells are often found within these cell clumps, such as secretory cells from the endocervic and low-grade squamous intraep-ithelial lesions (Lee et al 2011). Therefore, it is no surprise to see that the acquisition of multiple focal planes achieves better diagnostic performances than acquiring only one or a few focal planes (Evered and Dudding 2011;Wright et al 2013). Additionally, interpreting cellular objects requires the microscope to operate at high magnification (40×), preferably at resolutions close to the diffraction limit (∼0.2µm/pixel), where the DOF is reduced to around one micron (Castleman 1996).…”

Section: Introductionmentioning

confidence: 99%

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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Use of Digitally Stained Multimodal Confocal Mosaic Images to Screen for Nonmelanoma Skin Cancer

Lewin

Stevenson

et al. 2016

JAMA Dermatol

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IMPORTANCE Confocal microscopy has the potential to provide rapid bedside pathologic analysis, but clinical adoption has been limited in part by the need for physician retraining to interpret grayscale images. Digitally stained confocal mosaics (DSCMs) mimic the colors of routine histologic specimens and may increase adaptability of this technology.OBJECTIVE To evaluate the accuracy and precision of 3 physicians using DSCMs before and after training to detect basal cell carcinoma (BCC) and squamous cell carcinoma (SCC) in Mohs micrographic surgery fresh-tissue specimens.DESIGN This retrospective study used 133 DSCMs from 64 Mohs tissue excisions, which included clear margins, residual BCC, or residual SCC. Discarded tissue from Mohs surgical excisions from the dermatologic surgery units at

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Accuracy and perceptions of virtual microscopy compared with glass slide microscopy in cervical cytology

Cited by 86 publications

References 10 publications

A method for quantitative analysis of clump thickness in cervical cytology slides

A method for quantitative analysis of clump thickness in cervical cytology slides

Methods for rapid and high quality acquisition of whole slide images

Use of Digitally Stained Multimodal Confocal Mosaic Images to Screen for Nonmelanoma Skin Cancer

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