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Microplate Based Assays for the Study of Neurodegenerative Diseases

Neurodegenerative disease is a general term that encompasses any disease that is characterized degeneration of nerve cells leading to cognitive decline and problems with movement. Since nerve cells don’t normally regenerate these diseases are progressive and cannot be cured. Many neurodegenerative diseases like Alzheimer’s, Parkinson’s and prion diseases are characterized by a buildup of protein plaques that cut off nerve cell communication and are likely involved in the eventual death of these cells.

Thioflavin T (ThT) has long been the choice among scientist studying Alzheimer’s to detect amyloid fibrils that form the plaques in this disease. ThT exhibits a large enhancement in fluorescence emission upon binding to fibrils and has subsequently been found to be suitable for binding and thus detecting numerous other protein oligomers that form in a similar fashion.

preferred binding site of ThT to pleated sheets

Figure legend: Preferred binding site of ThT to b-pleated sheets From Wu, et al ‘Binding Modes of Thioflavin-T to the Single-Layer b-Sheet of the Peptide Self-Assembly Mimics’

Scientist studying prion diseases used our Omega microplate reader to develop an assay to detect prion infectivity of samples. The assay employs defined cycles of high intensity shaking and detects an increase in ThT as prion protein is converted to an infective form and oligomerization occurrs. The Omega platform proved to have the robust components needed to withstand the particular strains that this assay puts on the instrument. The assay, now known as RT-QuIC (real time quaking induced conversion) has been applied similarly to beta-amyloid for Alzheimer’s and alpha-synuclein for Parkinson’s.

Representative RT-QuIC data

( Image from CLARIO Omega Prion Brochure 2017 page 5 fig 2) Figure legend: Representative RT-QuIC data ThT fluorescence increases more rapidly with samples of high concentration

In response to RT-QuIC user needs we have made minor modifications to the Omega designed to ehance performance of this assay. These alteration are designed assure that reader / computer communication is maintained, prevent possible plate movement during shaking, improve ease of assay setup and improve shaking mechanism lifespan. Similar modifications are also available on the CLARIOstar®. So whether your lab needs dedicated instrumentation for RT-QuIC or have additional detection needs BMG has options that you can depend on.

FLUOstar Omega equipped to perform RT-QuIC

(Image from CLARIO Omega Prion Brochure 2017 page 2) Figure legend: FLUOstar Omega equipped to perform RT-QuIC

 

 

 

 

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Figure Legend: Modified plate carrier available on CLARIOstar® and Omega Plate carrier is designed to ensure plate is held securely in place during vigorous shaking necessary for RT-QuIC