Unique magnification 20, green Keratin 8/18 and reddish Keratin 5 immunostaining. HIS-74-1036-s008.avi (6.1M) GUID:?07AB14E9-E894-4D9B-96D5-1472A4440366 Video S9. S6. Z\stack of partial atrophy with an irregular acinar architecture of short blind\closing tubules and large areas lacking a basal cell coating. Initial magnifications 20, green Keratin 8/18 and reddish Keratin 5 immunostaining. HIS-74-1036-s006.avi (4.7M) GUID:?66257CE2-3D30-4F90-8CC5-D597277A4F78 Video S7. Three\dimensional rendering of Golgi\like atrophy, showing parallel AAI101 longitudinal ellipsoid glandular spaces resembling the Golgi\apparatus. Initial magnification 20, green Keratin 8/18 and reddish Keratin 5 immunostaining. HIS-74-1036-s007.avi (5.6M) GUID:?612F122C-230A-40B8-AA46-5AA4CF06EDB3 Video S8. Three\dimensional rendering of adenosis consisting of tubules without obvious acinar organization, closely mimicking the structure of Gleason pattern 3 prostate malignancy. Initial magnification 20, green Keratin 8/18 and reddish Keratin 5 immunostaining. HIS-74-1036-s008.avi (6.1M) GUID:?07AB14E9-E894-4D9B-96D5-1472A4440366 Video S9. Z\stack of high\grade prostate intraepithelial neoplasia with saccular architecture and abundant papillary protrusions. Initial magnification 20, green Keratin 8/18 and reddish Keratin 5 immunostaining. HIS-74-1036-s009.avi (8.2M) GUID:?FF54B653-23D8-4400-B3EF-078A4FCF2C0A Video S10. Z\stack of intraductal carcinoma of the prostate with gland spanning epithelial proliferations and irregular interconnecting luminal spaces. Initial magnification 20, green Keratin 8/18 and reddish Keratin 5 immunostaining. HIS-74-1036-s010.avi (10M) GUID:?02C987F3-B8F3-418B-AA31-5BB818B9C46B ? HIS-74-1036-s011.docx (17K) GUID:?2D06C03F-E5C7-4B66-8B2B-A3BDE8D52DF1 AAI101 Abstract Seeks Many glandular lesions can mimic prostate cancer microscopically, including atrophic glands, adenosis and prostatic intraepithelial neoplasia. While the characteristic histopathological and immunohistochemical features of these lesions have been well established, little is known about their three\dimensional architecture. Our objective was to evaluate the three\dimensional organisation of common prostate epithelial lesions. Methods and results 500?m\solid punches (and animal models, we have recently adapted the strategy for use about formalin\fixed, paraffin\embedded prostate cells. This allows 3D imaging of specific regions of interest.10 With this study we aimed to clarify the 3D architecture of the most common benign and precancerous prostate glandular lesions. Materials and Methods Case Selection A selection was made of archival formalin\fixed, paraffin\inlayed radical prostatectomy specimens from individuals who experienced undergone radical prostatectomy in the Erasmus Medical Center in Rotterdam between 2012 and 2017. These cells specimens were fixed in neutral\buffered formalin and regularly processed for histopathological evaluation. A urogenital pathologist (G.v.L.) indicated regions of interest on 5\m\solid H&E\stained cells slides. In total, 42 areas from 32 individuals were used. These areas included glands from the normal peripheral (fusions in adenosis.20, 21, 22, 23 Although adenosis is not a malignant proliferation, it may share aberrant activation of molecular pathways involved in 3D glandular development with low\grade PCa. A strong point of the current study was the use of intact cells samples from archival prostate specimens. This facilitated selection of specific regions\of\interest for 3D renderings. Imaging of small intact cells samples helps prevent alignment artefacts that could happen when using many consecutively cut and stacked slides. However, the use of fluorescently labelled antibodies to distinguish between luminal and basal epithelial cells restricts the potential sample volume to be visualized, owing to limited antibody penetration, especially in compact glandular constructions. Consequently, 3D imaging of larger areas was not feasible using AAI101 the current protocol, which is a disadvantage compared to additional 3D imaging techniques SGK2 such as sign up of serial sections.24 In addition, image analysis is time\consuming, and this meant that we were able to evaluate only a limited number of cells samples, not covering the entire range of benign prostate epithelial lesions. In conclusion, long\range confocal microscopy and optical cells clearing can be used to comprehensively visualise the unique 3D architecture of benign epithelial lesions of the prostate. While numerous lesions might mimic malignancy on routine cells slides, we found that their 3D architecture is mostly entirely different from PCa. Finally, 3D imaging can be helpful in elucidating the pathophysiology of benign prostate glandular lesions. Conflicts of interest The authors declare no.
- Remove supernatant and resuspend in 1 ml ice-cold HBSS+
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