Rapid histopathological evaluation of fresh, unfixed human tissue using optical sectioning microscopy would have applications to intraoperative surgical margin assessment. Microscopy with ultraviolet surface excitation (MUSE) is a low-cost optical sectioning technique using ultraviolet illumination which limits fluorescence excitation to the specimen surface. In this paper, we characterize MUSE using high incident angle, water immersion illumination to improve sectioning. Propidium iodide is used as a nuclear stain and eosin yellow as a counterstain. Histologic features of specimens using MUSE, nonlinear microscopy (NLM) and conventional hematoxylin and eosin (H&E) histology were evaluated by pathologists to assess potential application in Mohs surgery for skin cancer and lumpectomy for breast cancer. MUSE images of basal cell carcinoma showed high correspondence with frozen section H&E histology, suggesting that MUSE may be applicable to Mohs surgery. However, correspondence in breast tissue between MUSE and paraffin embedded H&E histology was limited due to the thicker optical sectioning in MUSE, suggesting that further development is needed for breast surgical applications. We further demonstrate that the transverse image resolution of MUSE is limited by the optical sectioning thickness and use co-registered NLM to quantify the improvement in MUSE optical sectioning from high incident angle water immersion illumination.
The assessment of protein expression in immunohistochemistry (IHC) images provides important diagnostic, prognostic and predictive information for guiding cancer diagnosis and therapy. Manual scoring of IHC images represents a logistical challenge, as the process is labor intensive and time consuming. Since the last decade, computational methods have been developed to enable the application of quantitative methods for the analysis and interpretation of protein expression in IHC images. These methods have not yet replaced manual scoring for the assessment of IHC in the majority of diagnostic laboratories and in many large-scale research studies. An alternative approach is crowdsourcing the quantification of IHC images to an undefined crowd. The aim of this study is to quantify IHC images for labeling of ER status with two different crowdsourcing approaches, image-labeling and nuclei-labeling, and compare their performance with automated methods. Crowdsourcing- derived scores obtained greater concordance with the pathologist interpretations for both image-labeling and nuclei-labeling tasks (83% and 87%), as compared to the pathologist concordance achieved by the automated method (81%) on 5,338 TMA images from 1,853 breast cancer patients. This analysis shows that crowdsourcing the scoring of protein expression in IHC images is a promising new approach for large scale cancer molecular pathology studies.
Experimental evidence supports a protective role of 25-hydroxyvitamin D (25(OH)D) in breast carcinogenesis, but epidemiologic evidence is inconsistent. Whether plasma 25(OH)D interacts with breast tumor expression of vitamin D receptor (VDR) and retinoid X receptor-alpha (RXR) has not been investigated. We conducted a nested case-control study in the Nurses’ Health Study, with 1,506 invasive breast cancer cases diagnosed after blood donation in 1989–90, 417 of whom donated a second sample in 2000–02. VDR and RXR expression were assessed by immunohistochemical staining of tumor microarrays (n=669 cases). Multivariate relative risks (RRs) and 95% confidence intervals (CIs) were calculated using conditional logistic regression.
Results
Plasma 25(OH)D levels were not associated with breast cancer risk overall (top (≥32.7ng/mL) vs. bottom (<17.2ng/mL) quintile RR=0.87, 95% CI (0.67–1.13), p-trend=0.21). 25(OH)D measured in summer (May–October) was significantly inversely associated with risk (top vs. bottom quintile RR=0.66, 95% CI (0.46–0.94), p-trend=0.01); winter levels (November–April) were not (RR=1.10, 95% CI (0.75–1.60), p-trend=0.64; p-interaction=0.03). 25(OH)D levels were inversely associated with risk of tumors with high expression of stromal nuclear VDR (≥30ng/mL vs. <30ng/mL RR (95% CI): VDR≥median=0.67 (0.48–0.93); VDR
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