With growing interest in leveraging in vitro models to reduce costs, increase throughput, and minimize ethical concerns throughout the early stages of drug discovery and development, microplates utilized in in vitro assays have become increasingly specialized. But, with the wide variety of microplate types on the market today, plate choice can be a daunting decision when setting up any new assay. Many considerations need to be taken into account such as culture approach (if any), throughput requirements, and analytical needs. Below we summarize a few of these considerations.
Immunofluorescence is a powerful tool that we utilize at Visikol to image whole tissues and 3D cell culture models. The imaging of large tissues can provide powerful quantitative insight into the sample of interest like protein colocalization, spatial information about cell viability, and penetration depth of test compounds.
In this blog post Dr. Tom Villani discusses loading and processing 3D confocal image stacks and performing volume measurements with ImageJ. This is crucial for processing data from tissues imaged in 3D with confocal or light sheet microscopy through the use of tissue clearing.
A major step in increasing the impact of these technologies is to improve throughput of screening. Successful genetic alteration or chemical resistance can be screened chemically, but morphological improvement will still be assessed via microscopy by skilled technicians (at least until machine learning can catch up). To assist with this effort we have developed the Visikol for Plant Biology reagent which allows for easy and high quality botanical microscopy through tissue clearing.
Primary cell spheroid cultures developed in this manner offer the potential to model complex diseases, previously only tractable in vivo. For example, a genetic modification or an environmental manipulation (such as a high fat diet or other stressor conditions) could be induced in an in vivo model and translated to an in vitro model by isolating relevant primary cells and culturing in 3D formats. To be clear, while the in vivo model would still require the use of animal subjects, the cells derived from a single or few subjects could be divided into hundreds of small models, enabling a higher throughput approach to a drug screen while still maintaining higher order relevance.
A common question for pharmaceutical companies developing therapeutics for neuroscience applications is the passage of small and large molecules across the blood brain barrier. Without this passage, these therapeutics will not reach their target and will thus not be efficacious. Therefore, many researchers have worked to develop in vitro models for the blood brain barrier that can mimic the pharmacokinetic properties in an inexpensive in vitro model.