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The challenge

The importance of molecular screening in drug discovery

The recent discovery that adult neurons are in fact capable of regeneration has triggered new hope for patients affected by these conditions. It has also attracted interest from the pharmaceutical industry in discovering new drugs that can positively affect neuron regeneration.

Image-based high throughput screening (HRT) is an important method in drug discovery to enable the analysis and modelling of neuron and neurite morphology. A neurite is any cell projection of the body of a neuron. The morphology of a neurite refers to the identification of its shape, structure, form and size. However, measuring changes in the morphology and characterising complex cellular structures such as neurite outgrowth are challenging tasks, especially with a dense image such as the one shown below.

A typical dense image with many “clumped” neurons and overlapped neurite structures. Left image shows a high degree of neurite branching complexity. Right image displays the segmented neurons and neurites using our software. ©Marjo Götte, Novartis Institutes for BioMedical Research

Several free and commercial high-content analysis software packages measure some basic features such as total neurite length and number of branching points. However, with only these measurements, neuronal morphologies that appear distinctly different to a human observer can be measured as identical by an automated classifier. Manually or semi-automatically doing this is a time-consuming, labour-intensive process, and the results are subjective and can undermine reliability.

Our response

An analysis algorithm for microscopy images

A robust and flexible high-content analysis (HCA) software package is essential for automated measurement and analysis of neuronal analysis. By measuring and analysing cellular responses in individual cells in a multi-well plate format, a flat plate with multiple "wells" used as small test-tubes, new compounds inducing neurite outgrowth can be identified. Hence, effective neurite tracing and measuring of changes in neuron morphology is crucial.

Our neurite analysis software is an automated solution for the quantification of neurite growth assays, investigative analytical techniques commonly used in target identification/validation and compound screening in drug discovery. This analysis algorithm can be used for analysis of both florescent and bright-field microscopy images.

This automated assay enables quantitation and correlation of neuron and neurite morphology. Analysis can range from simple neurite outgrowth measurement to complex analysis of extended neurite outgrowth and branching. Neurons are selected using a nuclear stain to locate the cell and a choice of neuron-specific stains or immunofluorescent labels to detect the cell body and neurite.

The results

Automating batch-processing images for rapid drug identification

Our software has been developed to automate the process of measuring and examining, analysing and cloning a complex image in seconds. It can be used to automatically batch-process tens of thousands of images produced from a high-content screening system. This can significantly promote productivity in the pharmaceutical industry for the rapid identification of active drug candidates from millions of compounds.

Through collaborative partnerships we have developed custom solutions for more complex neurite outgrowth assays, such as neurite detection and analysis in 2D images of co-cultures of neurons and support cells, as well as 3D neurite detection and analysis in 2-photon confocal 3D images in neurons.

Specifically designed for the drug development of neurological disorders, our Neurite Outgrowth Analysis algorithm has been licensed by PerkinElmer, a supplier of innovative tools and technologies for life sciences and pharmaceutical drug discovery, while the Neurite Outgrowth analysis algorithm has been adopted by 247 licenses in 2019 and more than 1,500 in total so far. PerkinElmer has also incorporated CSIRO's neurite outgrowth analysis algorithm to run its High Content Screening System, Opera™, and its data management analysis software, Columbus™.

Apart from licensing our cellular image analysis modules to commercial partners, we have developed a standalone software package, HCA-vision, which offers powerful functionalities for the analysis of cellular images. It features an easy to use graphical user interface, extensive reporting capabilities, and fast processing. The package is being used by bio-medical research institutes and pharma companies in Australia and overseas.

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