In the realm of neurodegenerative diseases, an unseen but very impactful battle is being waged at the molecular level. Central to this battle are misfolded proteins. These misfolded proteins are key players in the development of debilitating conditions like Alzheimer’s, Parkinson’s, and ALS.

Although we don’t fully understand these conditions, it seems that they’re all characterized by unique misfolded proteins or protein combinations which leads to the accumulation of protein deposits. These accumulations result in specific progressive neuronal pathological features and clinical symptoms​​. Understanding these misfolded proteins is crucial for developing effective treatments.

So how do we study them?

Microplate Readers

In the world of scientific innovations, not all heroes make the headlines. Some products have a profound impact, yet their presence often remains in the shadow of more glamorous technologies. Among these unsung heroes are multi-mode microplate readers – modest in size but monumental in their contribution to scientific advancement.

While multi-mode microplate readers may not be well known their role in propelling research across various fields is indispensable. They can be used to study the above-mentioned protein misfolding, but they do much more.

Multi-mode microplate readers are versatile laboratory instruments. They’re capable of performing various types of assays by combining different detection methods in a single device. These readers are designed to offer flexibility and efficiency in high-throughput screening, essential in research areas like drug discovery, biochemistry, and molecular biology. The “multi-mode” aspect refers to the ability of the reader to use different modes of detection, including:

1. Absorbance (Spectrophotometry): Measures the amount of light absorbed by a sample at specific wavelengths. This is commonly used for assays like ELISA, protein concentration, and cell growth analysis.
2. Fluorescence Intensity: Detects the intensity of light emitted by fluorophores in the sample after being excited by a specific wavelength of light. It’s widely used for DNA, RNA, and protein quantification, as well as many other applications.
3. Luminescence: Measures light emitted by the sample without any external light excitation, which is critical for assays involving bioluminescent or chemiluminescent reactions.
4. Time-Resolved Fluorescence (TRF): This technique, which involves measuring the delay between excitation and emission of light, is particularly useful in studying complex biological processes.
5. Fluorescence Polarization (FP): Used in binding studies and immunoassays, this method measures the change in the plane of polarized light as it passes through a fluorescent sample.
6. AlphaScreen/AlphaLISA: These are bead-based assays used for studying protein-protein interactions, enzyme activities, and receptor-ligand binding.
7. Förster Resonance Energy Transfer (FRET): This technique is used to measure the distance between two fluorophores, which can indicate molecular interactions.

No doubt, they’re pretty useful in different ways.

A versatile tool for science

The advantage of multi-mode microplate readers lies in their versatility. Researchers can switch between different assay types without needing multiple, dedicated instruments. This not only saves space and cost but also streamlines workflow in the laboratory.

When scientists need to perform multiple detection modes (as in, perform multiple times of measurements), they often turn to a multi-mode microplate reader. This tool also allows them to study different aspects of biological processes that enable scientists to accumulate data to help solve whichever problem they may be working on.

Microplate readers have greatly accelerated scientific research by allowing for rapid, automated analysis of many samples simultaneously, providing consistent and reproducible results. Their versatility and efficiency make them a staple in modern laboratory environments.

For instance, in the realm of genetics, multi-mode microplate readers are instrumental in experiments involving gene expression and regulation. A notable study involved using a microplate reader to understand the genetic basis of cystic fibrosis, leading to targeted therapies that significantly improve patient outcomes. The tools were also instrumental in the development of new drugs; and of course, during the recent pandemic, they played a key role.

Infectious diseases, such as COVID-19, have underscored the need for rapid, reliable testing and research tools. Multi-mode microplate readers have been at the forefront of this battle. During the COVID-19 pandemic, they played a crucial role in vaccine development and testing, enabling scientists to quickly evaluate vaccine efficacy against various virus mutations. This rapid response was vital in controlling the spread of the virus and saving countless lives.

Then, of course, there are neurological conditions.

Microplate readers and the brain

Microplate readers are valuable tools in studying Alzheimer’s and other neurological conditions due to their ability to perform high-throughput screening and quantitative analysis of various biological assays. Protein misfolding is just the start. These capabilities are crucial in understanding the complex molecular mechanisms underlying neurological disorders and in the development of potential therapeutic agents. Here’s how microplate readers contribute to this research:

1. Drug Discovery and Screening: Microplate readers allow for the rapid screening of thousands of compounds to identify potential drugs that can modify the course of neurological diseases. Researchers can test the effects of these compounds on cell viability, protein aggregation, enzyme activity, or receptor binding, all of which are relevant to conditions like Alzheimer’s.
2. Protein Aggregation Studies: In Alzheimer’s and other neurodegenerative diseases, the aggregation of specific proteins (like beta-amyloid and tau in Alzheimer’s) is a key pathological feature. Microplate readers can be used to quantify these protein aggregates using fluorescence or absorbance-based assays.
3. Cell Viability and Apoptosis Assays: Understanding how neurons die in neurodegenerative diseases is crucial. Microplate readers can assess cell viability, apoptosis (programmed cell death), and cytotoxicity in neuronal cells under various conditions, including exposure to neurotoxic agents or potential therapeutic compounds.
4. Calcium Signaling: Disrupted calcium signaling is implicated in many neurological conditions. Using fluorescent dyes that respond to calcium levels, microplate readers can monitor calcium signaling pathways in neurons.
5. Gene Expression Analysis: Changes in gene expression are common in neurological disorders. Fluorescence-based assays, such as quantitative PCR, can be performed on microplate readers to study these changes in a high-throughput manner.
6. Enzyme Activity Assays: Many neurological disorders involve dysfunctional enzymes. Microplate readers can measure the activity of these enzymes, aiding in understanding disease mechanisms and identifying potential drug targets.
7. Biomarker Detection: For diagnostic purposes, microplate readers can be used in immunoassays (like ELISA) to detect and quantify biomarkers associated with neurological diseases in biological samples.
8. Kinetic Studies: In neurodegenerative diseases, understanding the kinetics of certain pathological processes (like protein misfolding or aggregation) is important. Microplate readers can conduct time-course studies to observe these kinetics.
9. Neuroinflammation Research: They can be used to study neuroinflammation, a key component of many neurological disorders, by measuring the release of inflammatory cytokines or other markers of inflammation.

The future applications of microplate readers in neuroscientific research are far-reaching. With advancements in technology, these instruments will likely become even more sensitive, accurate, and versatile. This evolution will enable researchers to delve deeper into the molecular intricacies of neurological diseases, uncovering new pathways and targets for intervention.

The integration of artificial intelligence and machine learning in microplate reader technology could further revolutionize high-throughput screening, making it faster, more efficient, and capable of uncovering complex patterns and relationships in data that might otherwise go unnoticed.

This is just one tool out of many at the disposal of researchers; one tool you don’t read about on the news, but which is virtually indispensable to modern research.