616.234.5000

Histopathology

Pathology and Biorepository Core

The Core’s knowledgeable staff provide a range of histopathology services, in addition to protocol review and analytical expertise.

Service requests and pricing are managed through Agilent CrossLab (See Schedule Equipment and/or Request Services). Current CrossLab users can login here.

VARI staff that need a login (or further CrossLab support) should contact Lori Moon and MSU staff that need a login should contact Erica Lange. External users that need an CrossLab account/login can go here for account creation. After logging in, external users should click “list all cores” under the Core Facilities section in the left menu and scroll down to “Cores at Van Andel Institute.”

Histology services

The Core’s pathology services are led by Dr. Galen Hostetter, a board certified pathologist with more than 15 years of experience.

Automated hematoxylin and eosin (H&E) staining

H&E staining is the traditional stain for standard histology and morphology. The use of H&E stained slides to provide specific diagnosis based on morphology is a core skill of anatomic pathology.

Equipment:
Ventana Symphony Automated HE Stainer

Automated immunohistochemistry (IHC) staining (single and mulitplex ability)

IHC staining uses antibodies to detect the abundance, presence, and/or localization of specific proteins in a sample. This staining technique is critical to distinguish between diseases with similar morphology, as well as characterizing the molecular properties of certain cancers.

Antibodies are ordinarily monoclonal (mab), prepared primarily in mouse cells, or polyclonal (pcab), prepared in a variety of living hosts, including rabbit, goat, mouse or donkey, among others.

For multiplex approaches, the Core offers multiple stains that can mark the same or different analytes on the same tissue section. Two different antibodies or a combination of in situ hybridization (ISH) probe (DNA) and an antibody are applied to visualize expression and co-localized proteins at the tissue/cellular level. The ISH probe is used to determine species origin of harvested xenograft tissue. The automated immunostainer platform allows investigator design of additional multiplex strategies, with the potential of up to six markers simultaneously applied to a tissue section. This often requires histotechnician expertise, which the Core provides.

Equipment:
Dako Autostainer Link 48
Ventana Discovery Automated IHC Stainer

DNA in situ hybridization (ISH) staining and RNA scope

The fully automated Discovery Ultra immunostainer offers ISH protocols for chromogenic labeling of mRNA sequences in tissue sections. Various providers offer tested probes specific for mRNA of interest, which can be visualized with optimized denaturation and hybridization signals and by using a standard light microscope. Improved qualitative and quantitative signal enumeration is provided by VARI’s Vectra system.

Equipment:
Ventana Discovery Automated IHC Stainer

Tissue microarray

The purpose of the tissue microarray (TMA) is to represent (with small samplings of tissue through cores from source blocks) the histomorphological spectrum of the original surgical specimen. Therefore, construction of a TMA that will provide this appropriate representation necessitates careful case choice, case review, array design, technical skill at construction, and quality control. The advantage of a TMA slide is that hundreds of small samples are stained simultaneously on one slide, rather than hundreds of slides, thus saving costs of antibodies, stains, reagents and time.

Equipment:
Pathology Devices TMA Arrayer

Digital imaging

Digital slide scanners create electronic images of an entire microscope slide with superior image quality and speed. These high-quality electronic images can be transferred directly or hosted online for remote review.

Equipment:
Aperio Scanscope XT

Microscopy

Arcturus PixCell II laser capture microscope (LCM) is used for microdissection of histological samples into specific cell types. The isolated cells may be used for RNA/DNA analysis or other biochemical procedures.

Fluorescent labels maybe used to assist in the identification of the desired cell type.

A dual-headed Nikon microscope with a color CCD camera is available to image non-fluorescent samples at high resolution. The microscope also may be used to photograph immunohistochemistry (IHC) and special stains.

Equipment:
Arcturus PixCell II Laser Capture Microscope
Nikon Dual Headed Photomicroscope

Pathology services

Certified internal users should login to CrossLab and schedule service times through the system. For questions and pricing information, please contact Bree Berghuis at 616.234.5265.

Tissue fixation, processing and embedding

Fixation alters the tissue by stabilizing the proteins so that they are resistant to degredation. The aim of tissue processing is to remove the water from tissues and replace it with a medium that solidifies the tissue and allows for thin sections to be cut.

Equipment:
Sakura VIP5 Tissue Processor
Sakura VIP6 Tissue Processor
Leica Embedding Center

Paraffin sectioning/Cryosectioning

Thin sections of tissue are cut on rotary microtomes. Routine thickness for most sections is between 4 and 6 microns.

Equipment:
Leica RM2235
Leica RM2255
Leica CM1900 Cryostat