Histology Lab Service Core

The Histology Core is a core facility of the Indiana Center for Musculoskeletal Health and part of the Bone and Body Composition Core of the Indiana Clinical Translational Sciences Institute (CTSI). The Histology Core provides histological services for basic science (non-clinical) research. Both mineralized (plastic embedded) and soft tissue (paraffin embedded) specimens can be prepared by the facility.

Services

Plastic (methyl methacrylate) Embedding and Sectioning Service

This research facility offers processing of specimens through plastic (methyl methacrylate; MMA) embedding as well as thick sectioning of cortical bone, thin sectioning of trabecular (cancellous) bone, and staining of plastic sections.

The facility provides cover-slipped unstained sections for analysis of fluorochromes (dynamic histomorphometry) at no charge. Specialized stains (for static histomorphometry) are available. Goldner’s Trichrome (GT; osteoid, osteoblasts and osteoclasts), Hematoxylin and eosin (HE; routine assessment) and Toluidine Blue (TB; cement lines and cartilage) are available for both thick and thin sections. Tartrate-Resistant Acid Phosphatase (TRAP; osteoclasts) and Von Kossa / MacNeal (VKM; osteoid, osteoblasts and osteoclasts) are available for thin sections only.

Paraffin Embedding and Sectioning Service

The Histology Core offers de-mineralization of mineralized tissue (if necessary), processing of specimens through paraffin embedding, thin sectioning of paraffin-embedded tissue, and staining of paraffin sections.

For routine assessment, slides are stained with hematoxylin and eosin (HE; Harris hematoxylin, regressive method). Specialized stains are available, including Acidified Toluidine Blue (AcTB; for mast cells); Fast Green – Safranin O (FG/SO; for cartilage); Gomori’s Aldehyde Fuchsin, Wenk Modification (GAF; for pancreatic β cells, elastin, mast cells); Jones’ Periodic Acid-Silver Methenamine, AFIP Modification (JPA-ArM; for kidney glomerular capillary basement membrane); Masson’s Trichrome (MT; for collagen); Oil Red O (ORO; for lipids); Periodic Acid Schiff Procedure (PAS; for carbohydrates); Perls Iron Stain (PI); Tartrate-Resistant Acid Phosphatase (TRAP; for osteoclasts); Verhoeff-Van Giesen (VVG; for elastin); and Von Kossa Method for Mineral (VK).

Cryosectioning Service

The Histology Service Core offers frozen tissue sections. Slides can be stained for standard and specialized stains, or for immunohistochemistry. Samples for cryosections should be frozen by the investigator and submitted for sectioning by the core. Please contact Drew Brown to schedule the delivery of the tissues.

Visium Spatial Transcriptomics

If you are interested in next-generation molecular profiling, the Histology Core offers RNA-free sectioning for FFPE blocks in collaboration with the Center for Medical Genomics (CMG). We can section for any type of Visium slide, as well as produce "RNA scrolls" for you to test the RNA quality prior to the analysis. Please reach out to the CMG core first to schedule a time for the spatial transcriptomics analysis, and then we will make sure to complete the sectioning just prior to that date.

Materials for processing can be dropped off on the bench opposite the lab door (MS-5045G). All submissions require a completed work order form and request made through iLab.

Formalin-fixed tissue for either plastic or paraffin embedding should be submitted immersed in 70 percent ethanol (EtOH), not formalin. Specimens not submitted in histology cassettes will be charged for cassettes, if appropriate. N.B. Cassettes must be labeled with either pencil or solvent-resistant markers as listed below. Labeling done with Sharpies® or similar ink pens comes off during processing. Approved marking pens include Cancer Diagnostics Moist Marker Plus from Lab Safety Supply, HistoPrep Pen from Fisher Scientific, Klinipath KP Marker Plus from Mercedes medical, and Stafmark Pen from Statlab Medical Products.

Blocks for re-cutting and slides for staining can be submitted as is. The designated contact person will be notified by email when the work order is complete. For questions concerning submissions, email Dr. Lilian Plotkin.

To get started, you must register for an account:

1. Navigate to the core page: https://iu.ilab.agilent.com/service_center/show_external/5517/histology_services
2. In the upper-right-hand corner of the screen select the ‘Register using Indiana University Credentials’ option
3. You will be directed to an authentication page where you will need to enter your Indiana University credentials
4. Once you have entered your credentials, click the ‘Login’ button
5. You will be directed to an iLab Registration page where you will need to select your PI/Lab and verify your contact information.
6. Once your registration has been submitted, your PI will receive a notification that you have requested membership to their lab in iLab.  They will need to approve your membership and assign any accounts for your use.

Once you have been accepted into your PI’s lab and assigned accounts, you can create service requests.

1. Navigate to the core page: https://iu.ilab.agilent.com/service_center/show_external/5517/histology_services
2. At the upper right hand of the page ‘Sign In’ and then select the ‘Sign In using Indiana University Credentials’ option.
3. Enter your Indiana University and password, and sign in.
4. Select the Request Services tab and click on the ‘Request Service’ button next to the service of interest.
5. You will be asked to complete a form before submitting the request to the core.
6. Your request will be pending review by the core. The core will review your request and either Agree to the work or they will ask for more information if needed.