Tissue microarray - A new approach for the immunofluorescence based biomarker discovery in neurodegenerative diseases
Independent thesis Advanced level (degree of Master (Two Years)), 20 credits / 30 HE creditsStudent thesis
Alzheimer's disease (AD) and Lewy body dementia (DLB) are mong the most common form of dementia. Neurodegenerative diseases affect many people worldwide, and due to the lack in therapeutic targets, as well as biomarkers for diagnostics, there is a need for novel biomarkers for monitoring disease progression and especially early diagnose. A list of disease associated, brain enriched genes served as a starting point for the initiative phase in the search of a protein as a potntial biomarker. The method used was multiplex- immunofluorescence, in which a limitationtoday is the availability of primary antibodies raised in different host species. Here, heatinactivation as elution to eliminate cross-reaction of antibodies was investigated using tyramide signal amplification in an automated stainer. As proof of principle, sequential staining wasperformed on human cortex, with four rabbit antibodies targeting different morphological structures. There was no cross-rection between secondary antibodies and the protocol designed was used throughout the study. Protein distribution pattern was analysed on a tissue microarray(TMA) composed of human temporal cortex tissue obtained from 29 subjects; ten from patientsdiagnosed with AD, ten with DLB and nine controls, two cores from each case. IF was applied on proteins selected from the list provided, to assess how they relate to disease progression and evaluate TMA as approach. TMA was regarded as a useful tool for initiative step in screening for proteins as biomarkers, as long as representative tissue for cores is carefully selected.There are challenges important to address in using the sequential staining method and the TMA, but as a whole, a new approach for qualitative analysis of protein distribtuion was proposed. With thisapproach, the full potential of the antibodies generated within the HPA project, and other primary antibodies derived from the same species, can be combined with desired fluorophore within the same staining protocol, in a high throughput screening for proteins as a biomarker.
Place, publisher, year, edition, pages
Alzheimer's disease, multiplex, immunofluorescence, tissue microarray, brain, neuroscience
Engineering and Technology
IdentifiersURN: urn:nbn:se:kth:diva-163674OAI: oai:DiVA.org:kth-163674DiVA: diva2:801742