Single cell deformability cytometry using microfluidics

The changes in cellular mechanical properties are linked to several biological activities such as cell differentiation, cell proliferation and disease development. Conventional biomechanical tools are slow (~10 cells/hr) and laborious, which advocates a critical need for novel tools enabling high th...

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Main Author: Low, Xavier Jun Wei
Other Authors: Hou Han Wei
Format: Final Year Project (FYP)
Language:English
Published: Nanyang Technological University 2020
Subjects:
Online Access:https://hdl.handle.net/10356/141647
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author Low, Xavier Jun Wei
author2 Hou Han Wei
author_facet Hou Han Wei
Low, Xavier Jun Wei
author_sort Low, Xavier Jun Wei
collection NTU
description The changes in cellular mechanical properties are linked to several biological activities such as cell differentiation, cell proliferation and disease development. Conventional biomechanical tools are slow (~10 cells/hr) and laborious, which advocates a critical need for novel tools enabling high throughput single-cell mechanophenotyping. In this thesis, we report a novel microfluidic deformability cytometer which compasses the use of hydropipetting technique with viscoelastic fluid to induce cell deformation in a rapid and precise manner. The microchannel design consists of inertial-based asymmetric flow focusers and two cross junctions where fluid siphoning and hydropipetting occur continuously to induce flow-induced cell stretching and compression, respectively. As a proof of concept, polydimethylsiloxane (PDMS) microparticles of different stiffness were used to evaluate the sensitivity of the device. We first optimized the fabrication method (surfactant, vortex timing), and characterize particle size and yield using 2 different PDMS formulations (PDMS 184, PDMS 527). Next, we tested the deformability cytometer using the fabricated PDMS particles and showed that we can distinguish them based on deformability index. Finally, bladder carcinoma (HTB9) cells treated with and without paraformaldehyde (PFA) (a cell fixing agent) were tested, and significant morphological differences were observed on the stiffer PFA-treated HTB9 cells. Taken together, the results demonstrated the developed technology is high throughput (100 cells/sec) with a large dynamic range of cell stiffness (5.0 kPa to 1.7 MPa), and can be further developed as a label-free single cell technique for rapid and point-of-care diagnostics.
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spelling ntu-10356/1416472023-03-04T19:40:23Z Single cell deformability cytometry using microfluidics Low, Xavier Jun Wei Hou Han Wei School of Mechanical and Aerospace Engineering Thales at NTU Joint Research Laboratory hwhou@ntu.edu.sg Engineering::Mechanical engineering The changes in cellular mechanical properties are linked to several biological activities such as cell differentiation, cell proliferation and disease development. Conventional biomechanical tools are slow (~10 cells/hr) and laborious, which advocates a critical need for novel tools enabling high throughput single-cell mechanophenotyping. In this thesis, we report a novel microfluidic deformability cytometer which compasses the use of hydropipetting technique with viscoelastic fluid to induce cell deformation in a rapid and precise manner. The microchannel design consists of inertial-based asymmetric flow focusers and two cross junctions where fluid siphoning and hydropipetting occur continuously to induce flow-induced cell stretching and compression, respectively. As a proof of concept, polydimethylsiloxane (PDMS) microparticles of different stiffness were used to evaluate the sensitivity of the device. We first optimized the fabrication method (surfactant, vortex timing), and characterize particle size and yield using 2 different PDMS formulations (PDMS 184, PDMS 527). Next, we tested the deformability cytometer using the fabricated PDMS particles and showed that we can distinguish them based on deformability index. Finally, bladder carcinoma (HTB9) cells treated with and without paraformaldehyde (PFA) (a cell fixing agent) were tested, and significant morphological differences were observed on the stiffer PFA-treated HTB9 cells. Taken together, the results demonstrated the developed technology is high throughput (100 cells/sec) with a large dynamic range of cell stiffness (5.0 kPa to 1.7 MPa), and can be further developed as a label-free single cell technique for rapid and point-of-care diagnostics. Bachelor of Engineering (Mechanical Engineering) 2020-06-10T00:59:17Z 2020-06-10T00:59:17Z 2020 Final Year Project (FYP) https://hdl.handle.net/10356/141647 en A187 application/pdf Nanyang Technological University
spellingShingle Engineering::Mechanical engineering
Low, Xavier Jun Wei
Single cell deformability cytometry using microfluidics
title Single cell deformability cytometry using microfluidics
title_full Single cell deformability cytometry using microfluidics
title_fullStr Single cell deformability cytometry using microfluidics
title_full_unstemmed Single cell deformability cytometry using microfluidics
title_short Single cell deformability cytometry using microfluidics
title_sort single cell deformability cytometry using microfluidics
topic Engineering::Mechanical engineering
url https://hdl.handle.net/10356/141647
work_keys_str_mv AT lowxavierjunwei singlecelldeformabilitycytometryusingmicrofluidics