Histology & Molecular Pathology Core

The overall goals of the Histology and Molecular Pathology Core are:

To provide a common facility and expertise for the broad spectrum of morphologic studies for interpretation of models of kidney disease and response to interventions
To provide tissue samples from kidney disease models for assessment of expression patterns of novel target molecules
To develop novel molecular approaches that enhance existing morphologic techniques to assess specific molecules in injured tissue, and to interpret their significance based on the spatial localization and relationship to patterns of injury.

The Histology and Molecular Pathology Core offers processing, embedding and staining of frozen and paraffin-embedded tissues and cell culture to study normal and different AKI and CKD models.

We analyze these tissue sections that have been stained with hematoxylin and eosin (H&E), periodic acid-schiff (PAS), trichrome, Sirius red and/or immunohistochemistry for morphological injury and response to various interventions.

We have worked with about 12 different pharmaceutical companies. At least 25 investigators from outside institutions and from Vanderbilt departments and divisions, including Adult Nephrology, Pediatric Nephrology, Cardiology, Pharmacology and Pathology, have utilized our histology services.

Core Assays and Service

Processing/Embedding/Sectioning

Specific objective:

To provide appropriate processing of tissue, including freezing, fixation in various media, paraffin-embedding and sectioning
To provide expert advice on preparation of tissue for electron microscopic studies

Available service:

Communication with investigators through expert Core personnel on how to prepare tissue. Examples of this include clamping of renal pedicle vessels before terminal blood drawing, procedures for laparoscopic renal biopsies, optimal sectioning, and allocation of tissues for various molecular and morphologic needs
Processing tissue samples for embedding and sectioning

Staining and Molecular Histology

Specific objective:

To provide routine H&E and special stains as needed
To provide immunohistochemistry (IHC) for selected panels of tested and validated antibodies, multiplex staining
To provide in situ hybridization (ISH), using the state of the art novel senstitive RNAscope approach
To provide expert advice on spatial proteinomics/transcriptomics

Available service:

Processing tissue samples for various stains (Table 1)
Provide a selected array of IHC for validated and tested antibodies
Spatial transcriptomics approach based of specific project needs (Figure 1)

Table 1:

Stain	Staining Properties	Key Use
Hematoxlyn and Eosin (H&E)	Nuclei and cytoplasm	Tubular cell injury, inflammation
Jones silver stain	Basement membrane/matrix	GBM changes/matrix
Masson's Trichrome	Collagen	Fibrosis
Sirius red	Collagen	Fibrosis (polarized for optimal quantitation)
Periodic acid-schiff (PAS)	Glycoproteins	Arterial hyalinosis, matrix increase
Elastin	Elastic fibers	Arterial injury
Von Kossa	Phosphate (part of calcium phosphate)	Calcium-phosphate deposits
Alizarin red	Calcium	Calcium deposits
Congo red	Intercalates with β-pleated sheets	Detection of amyloid

Figure 1:

Tissue Array

Specific objective:

To provide archival tissue samples from tissue arrays of various disease phenotypes, treated or nontreated, to detect key novel molecules of interest

Available service:

Mouse tissue microarrays (TMAs) - contains 144 cores of kidney tissue samples and 12 cores of normal samples. Including sample from C57BL/6J, DBA/2J, FVB/NJ, MRL/MpJ, A/J, KK/HlJ, eNOS-/- x db/db, nephrin COX-2 transgenic, floxed AADC, floxed 11βHSD2, AT1a and AT2 receptor knockout mice
Human diabetic nephropathy tissue microarrays (hDN-TMA) - contain 126 cores of kidney tissues with diabetic nephropathy (42 different patients) and 30 cores of kidney tissues with minimal or no abnormality from donor biopsies (10 donors)

Morphology Analysis

Specific objective:

To provide expert analysis of kidney tissues submitted to the Core
To provide expert advice and services for basic morphometry of key aspects of electron microscopy (EM) samples
To provide morphometric analysis by using artificial intelligence (Figure 2)

Available service:

Semi-quantification of glomerular injury, acute and chronic tubular injury, and vascular injury by LM
Quantitation of IHC
EM quantitation includes GBM width (measured directly and by harmonic mean), ultrastructural alteration of GBM composition, podocyte injury (e.g. extent of foot process effacement, slit diaphragm density per unit length of GBM/capillary loop), endothelial cell injury (e.g. loss of fenestrations), increase in mesangial matrix formation, and presence of immune complexes
Artificial intelligence assessment of key parameters of CKD, for instance interstitial fibrosis, tubular atrophy, global glomerulosclerosis

Figure 2:

Pricing Information

The service fees are set to recover a portion of the true costs of operation, including:

Service contracts (e.g. for tissue processor, electron microscope, microtomes)
Disposables, such as materials necessary for staining, glass slides, blades
Computer software for digital imaging and record maintenance
Staff time for routine hardware and software maintenance
Minor upgrades to equipment and software.

Please contact Core leadership for pricing.

Histology and Molecular Pathology Core

Histology & Molecular Pathology Core

Core Assays and Service

Processing/Embedding/Sectioning

Staining and Molecular Histology

Tissue Array

Morphology Analysis

Pricing Information

Core Leadership

Agnes Fogo, MD

Haichun Yang, MD, PhD

Ming-Zhi Zhang, MD

Vanderbilt O’Brien Kidney Center

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