Three-dimensional imaging of single isolated cell nuclei using optical projection tomography

Fauver, Mark; Seibel, Eric J.; Rahn, John; Meyer, Michaël; Patten, Florence W.; Neumann, Thomas; Nelson, Aimee J.

doi:10.1364/opex.13.004210

Cited by 135 publications

(112 citation statements)

References 22 publications

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“…Recent technical developments have made it possible to combine flow cytometry with CT on the cellular level, allowing for cytometry in 3 dimensions. 32 In addition, it has been demonstrated that automated cancer cell recognition based on 3-dimensional measurements is superior to 2-dimensional (slide-based) cell analysis.…”

Section: Discussionmentioning

confidence: 99%

Premalignant and malignant cells in sputum from lung cancer patients

Neumann

Meyer²,

Patten³

et al. 2009

Cancer Cytopathology

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BACKGROUND:The objective of this study was to assess the frequency of premalignant and malignant cells in sputum from patients with lung cancer and to measure the dependence of these cells on cancer stage, histologic type, tumor size, and tumor location.METHODS:This analysis included 444 patients with lung cancer. First, all patients were asked to produce sputum spontaneously; then, they underwent sputum induction. Slide preparations of the sputa were screened for the presence of abnormal cells.RESULTS:Of all patients with lung cancer who had produced adequate specimens, 74.6% had sputum that was positive for premalignant or worse cells, whereas 48.7% had sputum that was positive for malignant cells alone. Surprisingly, the presence of premalignant or worse cells in sputum depended only moderately on disease stage (82.9% of stage IV cancers vs 65.9% of stage I cancers), tumor size (78.6% of tumors >2 cm vs 64.7% of tumors ≤2 cm), and location (83.3% of central lesions vs 68% of peripheral lesions) and was found to be independent of histologic tumor type (78.4% of squamous cell carcinomas vs 71.5% of adenocarcinomas, 74.5% of small cell carcinomas, and 75% of large cell carcinomas).CONCLUSIONS:The findings of the current study suggested the important potential of sputum cytology for lung cancer detection and risk assessment across all stages, histologic types, tumor sizes, and locations. However, the high sensitivities in this study were achieved with a level of scrutiny not feasible in the laboratory routine. The diagnostic potential of sputum cytology may be exploited better through the standardization and automation of sputum preparation and analysis. Cancer (Cancer Cytopathol) 2009. © 2009 American Cancer Society.

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Section: Discussionmentioning

confidence: 99%

Premalignant and malignant cells in sputum from lung cancer patients

Neumann

Meyer²,

Patten³

et al. 2009

Cancer Cytopathology

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“…OPT has been shown to reveal superb sectioning ability, useful for reconstructing vertebrate embryos and in the examination of the 3D anatomy of developing organs [222,224,225] (see Figure 18); modelling of early human brain development [226]; 3D imaging of isolated cell nuclei [227]. With regard to contributions this technique has made to the knowledge of flow and related dynamics, OPT has been shown to be applicable in studies of gas-solids flow to analyse particle concentration [228]; general flow rate analyses [229,230]; cell tracing [231]; and the spatiotemporal analysis of zebrafish [232].…”

Section: Optical Projection Tomographymentioning

confidence: 99%

‘Go with the flow ’: A review of methods and advancements in blood flow imaging

Daly

Leahy

2012

Journal of Biophotonics

108

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Physics has delivered extraordinary developments in almost every facet of modern life. From the humble thermometer and stethoscope to X‐Ray, CT, MRI, ultrasound, PET and radiotherapy, our health has been transformed by these advances yielding both morphological and functional metrics. Recently high resolution label‐free imaging of the microcirculation at clinically relevant depths has become available in the research domain. In this paper, we present a comprehensive review on current imaging techniques, state‐of‐the‐art advancements and applications, and general perspectives on the prospects for these modalities in the clinical realm. (© 2013 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)

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“…The system was built on a widefield optical microscope (Olympus IX71) with a customdesigned microcapillary-based rotation stage. 25 Since OPTM can do multimodal imaging, both absorption as well as fluorescence imaging were performed on the same instrument. 26 For absorption imaging, the light from the halogen lamp (QTH) is filtered by a diffuser and a color filter.…”

Section: Feulgen Staining the Feulgen Staining Technique Was Per-mentioning

confidence: 99%

“…OPTM generates 3-D, submicron, isometric, and high-resolution images of nuclear features. 25 Since the entire volume of the nucleus is measured, there is less uncertainty in the measure of total DNA content as every voxel is used to calculate the integrated optical density. Variations in perspective of the nucleus from 2-D images are eliminated in a single OPTM image.…”

Section: Introductionmentioning

confidence: 99%

Three-dimensional DNA image cytometry by optical projection tomographic microscopy for early cancer diagnosis

Agarwal

Biancardi

Patten³

et al. 2014

J. Med. Imag

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Abstract. Aneuploidy is typically assessed by flow cytometry (FCM) and image cytometry (ICM). We used optical projection tomographic microscopy (OPTM) for assessing cellular DNA content using absorption and fluorescence stains. OPTM combines some of the attributes of both FCM and ICM and generates isometric highresolution three-dimensional (3-D) images of single cells. Although the depth of field of the microscope objective was in the submicron range, it was extended by scanning the objective's focal plane. The extended depth of field image is similar to a projection in a conventional x-ray computed tomography. These projections were later reconstructed using computed tomography methods to form a 3-D image. We also present an automated method for 3-D nuclear segmentation. Nuclei of chicken, trout, and triploid trout erythrocyte were used to calibrate OPTM. Ratios of integrated optical densities extracted from 50 images of each standard were compared to ratios of DNA indices from FCM. A comparison of mean square errors with thionin, hematoxylin, Feulgen, and SYTOX green was done. Feulgen technique was preferred as it showed highest stoichiometry, least variance, and preserved nuclear morphology in 3-D. The addition of this quantitative biomarker could further strengthen existing classifiers and improve early diagnosis of cancer using 3-D microscopy.

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Three-dimensional imaging of single isolated cell nuclei using optical projection tomography

Cited by 135 publications

References 22 publications

Premalignant and malignant cells in sputum from lung cancer patients

Premalignant and malignant cells in sputum from lung cancer patients

‘Go with the flow ’: A review of methods and advancements in blood flow imaging

Three-dimensional DNA image cytometry by optical projection tomographic microscopy for early cancer diagnosis

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