Histological and Imaging Protocols for Tissue Analysis

Tissues were fixed in formalin, paraffin embedded, and sectioned at 5 microns, and were stained with hematoxylin and eosin (H&E), or prepared for immunostaining as previously described.[10 (link), 26 , 27 (link)] Images were captured on a Nikon Eclipse NI-U with a DS-QI1 or DS-Ri1 camera and NIS Elements software, and adjusted in Adobe Photoshop. Antibodies were as follows: rabbit anti-Krt10, chicken anti-Krt5, mouse anti-Krt8, and rabbit anti-Ki-67. Alexafluor 488, 546, and 647 secondary antibodies were from Thermo Fisher Scientific. Whole-mount in situ hybridization was performed on embryos with digoxigenin-labeled riboprobes, as described.[28 (link)] β-galactosidase staining was carried out as described[29 ]. Whole mount images were captured on a Leica MZ16 stereomicroscope and QImaging 5.0 RTV digital camera.

Partial Protocol Preview
This section provides a glimpse into the protocol.
The remaining content is hidden due to licensing restrictions, but the full text is available at the following link: Access Free Full Text.

Yang C.S., Melhuish T.A., Spencer A., Ni L., Hao Y., Jividen K., Harris T., Snow C., Frierson H., Wotton D, & Paschal B.M. (2017). The Protein Kinase C Super-family Member PKN is Regulated by mTOR and Influences Differentiation During Prostate Cancer Progression. The Prostate, 77(15), 1452-1467.

Publication 2017

Eclipse NI-U DS-QI1 DS-Ri1

Antibodies Camera Chicken Digoxigenin Embryos Eosin Formalin In situ hybridization Krt10 Krt5 Krt8 Mouse Paraffin Rabbit Tissues β galactosidase

Corresponding Organization : University of Virginia

Top 5 similar protocols

Variable analysis

independent variables

Tissue fixation method (formalin)
Paraffin embedding
Sectioning thickness (5 microns)
Staining method (H&E, immunostaining)
Antibodies used (rabbit anti-Krt10, chicken anti-Krt5, mouse anti-Krt8, rabbit anti-Ki-67)
Whole-mount in situ hybridization with digoxigenin-labeled riboprobes
β-galactosidase staining

dependent variables

Images captured on Nikon Eclipse NI-U microscope with DS-QI1 or DS-Ri1 camera
Image adjustment in Adobe Photoshop

control variables

Not explicitly mentioned

positive controls

Not specified

negative controls

Not specified

Annotations

Based on most similar protocols

Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.

As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!