Tailored high throughput microarray technique
NeuroValida uses tailored high throughput human brain tissue microarray technique to establish localisation of putative drug targets, the protocols for which are published in high-ranking journals (Waldvogel HJ, et al. Nature Protocols 1:2719-2732, 2006; Singh-Bains et al. Nature Protocols 16:2308-2343, 2021). The technique employs well characterised adult human brain tissue microarray slides containing up to 60 regions of interest (or 60 cases) effectively reducing experimental error and eliminating reagent variability and time delays between samples for drug discovery and validation. The tissue microarray slides are constructed from well characterized human brain available from the Neurological Foundation Human Brain Bank whose pathology has been verified independently by a board-certified pathologist and use a core biopsy of brain tissue approximately 2 mm in diameter set into a wax block. The array contains either different regions of the human brain or different brain specimens or a combination of the two. Once the array is made, sections typically 5-10 µm in thickness can be cut from the array using standard histological techniques. The sections containing 60 brain samples then undergo immunohistochemistry or a similar process. All brain samples are processed on a single slide and there is no variation in the processing from one case or region to the next therefore allowing for high reproducibility. Once stained, the slide is scanned at high resolution and MetaMorph® journals are used for data analysis (number of cells, cell size, neurite length, neurite number, staining density changes, area of staining etc.). The data is then used to feedback any changes that are evident with respect to the target of interest.
Available Tissue Microarrays
Available tissue microarrays focus on Alzheimer’s disease, Parkinson’s disease, Huntington’s disease, and motor neurone disease among others, to provide data on disease-specific markers. For example, our Alzheimer’s disease arrays have data on loads for tau, amyloid, microglia, astrocytes, etc. allowing us to correlate proteins of interest with key pathological features of Alzheimer’s disease. Our brain tissues are also exome sequenced allowing for availability of rich genetic data sets along with Braak, ARP and ApoE scores, in addition to well characterised loads for tau, amyloid, microglia, astrocytes, etc for each tissue microarray. This means that we can correlate proteins of interest with key pathological features of neurodegenerative diseases. We also supply matched neurologically normal controls for each of the neurological conditions and can include a selection of appropriate animal brain cores in each tissue microarray to reference concurrent research activities. We recognize that each customer has different needs and we work towards tailoring a solution that is best for you.
Automated Image Acquisition
We use the VSlide and Olympus SLIDEVIEW VS200 research slide-scanner to acquire images from tissue microarrays and then feed these images into our automated (high content) image analysis program.
Automated Image Analysis
We use MetaMorph® to perform automated high content image analysis which extracts both simple and complex features from the acquired images of the drug target, including cellular and sub-cellular localisation and many other features. Data is interpreted by the Neurovalida research team, who have a combined 60 years of expertise studying the anatomy and chemistry of the human brain.
We use ultrastructural studies to determine where proteins of interest might be in a cell using contrast enhanced methodology, immunostaining methods and gold beads of various sizes for colocalization studies. We provide images and a report of what we find using electron microscopy.