TAU Protein cell characterisation towards establishing microfluidic platforms

A dielectrophoresis (DEP) study on Tau-441 protein has been performed for lab-on-a-chip bioparticle manipulation investigation. In this work, a study of bioparticles manipulation which targeted on recombinant human Tau-441 protein under the gold rectangular shape microelectrode platform has been con...

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Main Author: Da En, Zuriel Shee
Format: Thesis
Published: 2022
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spelling my-mmu-ep.115372023-07-18T01:48:01Z TAU Protein cell characterisation towards establishing microfluidic platforms 2022-09 Da En, Zuriel Shee QC501-(721) Electricity A dielectrophoresis (DEP) study on Tau-441 protein has been performed for lab-on-a-chip bioparticle manipulation investigation. In this work, a study of bioparticles manipulation which targeted on recombinant human Tau-441 protein under the gold rectangular shape microelectrode platform has been conducted with employing dielectrophoresis (DEP) technique to investigate and characterise this Tau-441 protein which is a protein in the brain neurons that is associated with Alzheimer’s disease (AD). Series of characterisation of recombinant human Tau-441 protein has been conducted. The morphology and structure of the Tau-441 protein have been examined with optical microscopy and fluorescence microscopy analysis with an average length of 206.62µm. Through Fourier-Transform Infrared Spectroscopy, Tau-441 protein was misfolded into pair helical filaments (PHF) and neurofibrillary tangles (NFTs) forms and 84.28% of its secondary structure is made up of β-Sheet structure. An electrochemical impedance spectroscopy was performed to identify the Tau-441 dielectric properties, the average of conductivity of Tau-441 protein calculated is 1.046 S/m. A parametric study on the rectangular shape microelectrode was conducted by using COMSOL® simulation analysis, the optimum microelectrode design parameters studied is a 130µm × 130µm rectangular shape gold microeletrode and was fabricated by using photolithography technique. A DEP test on model yeast cells was conducted and manipulated with a positive DEP in 60 second at 5 peak to peak voltage (Vpp) with 5MHz frequency. Next, two DEP configurations experiments were conducted on Tau-441 protein which were suspended in deionised water (DIW) and artificial cerebrospinal fluid (CSF) which is the brain fluid. Both experiments shown a negative DEP response at 5V with 10MHz. Lastly, DEP separation experiment was conducted on the combination of model yeast cells representing AD infected brain cells and Tau-441 protein which is the AD associated protein in the CSF medium. A successful separation of model yeast cells and Tau-441 protein mixture was achieved at 5Vpp with 10MHz. Overall, this research has successfully trapped and separate Tau-441 protein aggregates with model yeast cells by using dielectrophoresis technique with a separation manipulation efficiency up to 90%. The knowledge from these investigations has brought significant insight for research related to establishing dielectrophoresis microfluidic brain-on-chip platforms relevant to Alzheimer’s disease. 2022-09 Thesis http://shdl.mmu.edu.my/11537/ http://erep.mmu.edu.my/ masters Multimedia University Faculty of Engineering and Technology (FET) EREP ID: 10866
institution Multimedia University
collection MMU Institutional Repository
topic QC501-(721) Electricity
spellingShingle QC501-(721) Electricity
Da En, Zuriel Shee
TAU Protein cell characterisation towards establishing microfluidic platforms
description A dielectrophoresis (DEP) study on Tau-441 protein has been performed for lab-on-a-chip bioparticle manipulation investigation. In this work, a study of bioparticles manipulation which targeted on recombinant human Tau-441 protein under the gold rectangular shape microelectrode platform has been conducted with employing dielectrophoresis (DEP) technique to investigate and characterise this Tau-441 protein which is a protein in the brain neurons that is associated with Alzheimer’s disease (AD). Series of characterisation of recombinant human Tau-441 protein has been conducted. The morphology and structure of the Tau-441 protein have been examined with optical microscopy and fluorescence microscopy analysis with an average length of 206.62µm. Through Fourier-Transform Infrared Spectroscopy, Tau-441 protein was misfolded into pair helical filaments (PHF) and neurofibrillary tangles (NFTs) forms and 84.28% of its secondary structure is made up of β-Sheet structure. An electrochemical impedance spectroscopy was performed to identify the Tau-441 dielectric properties, the average of conductivity of Tau-441 protein calculated is 1.046 S/m. A parametric study on the rectangular shape microelectrode was conducted by using COMSOL® simulation analysis, the optimum microelectrode design parameters studied is a 130µm × 130µm rectangular shape gold microeletrode and was fabricated by using photolithography technique. A DEP test on model yeast cells was conducted and manipulated with a positive DEP in 60 second at 5 peak to peak voltage (Vpp) with 5MHz frequency. Next, two DEP configurations experiments were conducted on Tau-441 protein which were suspended in deionised water (DIW) and artificial cerebrospinal fluid (CSF) which is the brain fluid. Both experiments shown a negative DEP response at 5V with 10MHz. Lastly, DEP separation experiment was conducted on the combination of model yeast cells representing AD infected brain cells and Tau-441 protein which is the AD associated protein in the CSF medium. A successful separation of model yeast cells and Tau-441 protein mixture was achieved at 5Vpp with 10MHz. Overall, this research has successfully trapped and separate Tau-441 protein aggregates with model yeast cells by using dielectrophoresis technique with a separation manipulation efficiency up to 90%. The knowledge from these investigations has brought significant insight for research related to establishing dielectrophoresis microfluidic brain-on-chip platforms relevant to Alzheimer’s disease.
format Thesis
qualification_level Master's degree
author Da En, Zuriel Shee
author_facet Da En, Zuriel Shee
author_sort Da En, Zuriel Shee
title TAU Protein cell characterisation towards establishing microfluidic platforms
title_short TAU Protein cell characterisation towards establishing microfluidic platforms
title_full TAU Protein cell characterisation towards establishing microfluidic platforms
title_fullStr TAU Protein cell characterisation towards establishing microfluidic platforms
title_full_unstemmed TAU Protein cell characterisation towards establishing microfluidic platforms
title_sort tau protein cell characterisation towards establishing microfluidic platforms
granting_institution Multimedia University
granting_department Faculty of Engineering and Technology (FET)
publishDate 2022
_version_ 1776101408155631616