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Since the Pathology Services Core is a core research facility providing a range of histology and imaging services, including complete tissue-to-data workflow, it is important that submitters follow good laboratory practices (GLP) in tissue handling, cell culture (if applicable), and record-keeping. This will ensure the data derived from these biological specimens will be of the highest quality. If you have any questions regarding the guidelines listed below, please contact PSC by telephone at 919-843-3409 or e-mail nanan@med.unc.edu.

Detecting fluorescence

For target detection on most of our fluorescence platforms (especially the ScanScope FL), a counterstain (e.g., DAPI/Hoechst) that is uniformly distributed across the tissue is essential for proper focusing. Alternatively, it is possible to stain serial sections for IHC (e.g., DAB detection) in order to identify the marker of interest (MOI). Such a slide can be scanned on our bright field scanner (ScanScope XT). Then, the MOI can be located more easily for the fluorescence scan. For multiplex target detection, we recommend the use of fluorophores that have maximum spectral separation between channels such as DAPI/Hoechst (blue channel), Alexa 488/FITC/GFP (green channel), Alexa 555/Cy3/RFP (orange channel), and Alexa 647/Cy5 (red channel) or equivalent reporter molecules.

To avoid residual glass artifacts that appear with fluorescence, gently rinse coverslips with deionized water immediately before mounting them. Please do not use non-drying mounting media (e.g., Vectashield or other glycerol-based mountants) as this has a tendency to leak (even from sealed coverslips). After mounting medium is fully cured, seal the coverslip to the slide with clear nail polish. Otherwise, the coverslip may move during handling, which can damage the tissue. Also, note that aqueous mounting media can slowly evaporate and shrink over time if coverslips are not sealed. This can be a problem for thick tissue sections. PSC currently uses ProLong Gold mountant (https://www.thermofisher.com/order/catalog/product/P36930) for fluorescence slides. Some of our clients have had good results with Fluoro-gel (https://www.emsdiasum.com/microscopy/technical/datasheet/17985.aspx). Avoid the use of Vectashield HardSet mountant as this can cause significant background fluorescence in the DAPI channel.

Good Input = Good Output

If customers submit slides with improper staining, smudges, mounting medium smears, or other poor quality preparation, PSC staff may spend extra time cleaning and/or remounting (if possible), and the Principal Investigator will be billed for the extra work. In rare cases, the slides cannot be safely scanned and will be returned to the P.I.

Regarding Access to Digital Slides in the eSlide Manager Database

The digital slide data stored on tpl-spectrum.med.unc.edu is considered the property of the data owner, i.e., the Principal Investigator or other party so named in the Research Agreement / IRB document on file with the Translational Pathology Laboratory. Therefore, only the data owner (P.I.) can grant access to this data in accordance with UNC policy. If a P.I.’s collaborator or lab member needs access to this data, the P.I. must explicitly authorize PSC to give this access by providing the data user’s name, UNC e-mail address, and work phone number to PSC management (look under “Contact Us” on the Home page). When requesting access for another person, the P.I. should also specify the level of access as either “Full Control” or “Read Only.” Full Control access grants the user the ability to edit and annotate digital slides in eSlide Manager. Read Only access allows only viewing of the slides. If no access level has been specified, it will be set to “Full Control” by default. Once PSC has approved the request, the new eSlide Manager account will be created, and instructions for logging in will be sent to the new user. Note that eSlide Manager accounts expire after six months, but can be easily renewed on demand. NOTE – PSC complies fully with all institutional policies regarding the security of Protected Health Information (PHI). If any digital slides contain PHI, only individuals named in the P.I.’s IRB documents will be permitted access to it.