THESIS
2018
xix, 108 pages : illustrations ; 30 cm
Abstract
Cells are the most fundamental structural and functional units of life. It was first discovered
and named by Robert Hooke in 1665, since when numerous biologists have been dedicated to
uncover its mysteries. In most conventional cellular bioassays, such as Western blotting, real-time
PCR, ELISA, etc., cell samples containing numerous individual cells are normally treated
as homogenous and analyzed in bulk level. However, in recent decades, advances in different
fields of biology have found that cellular heterogeneity widely exists in all cell populations.
Analysis in bulk assays may not accurately reflect true biology of individual cells. The studies
in single-cell level help to reveal critical secrets concealed in conventional population-averaged
measurements and provide valua...[
Read more ]
Cells are the most fundamental structural and functional units of life. It was first discovered
and named by Robert Hooke in 1665, since when numerous biologists have been dedicated to
uncover its mysteries. In most conventional cellular bioassays, such as Western blotting, real-time
PCR, ELISA, etc., cell samples containing numerous individual cells are normally treated
as homogenous and analyzed in bulk level. However, in recent decades, advances in different
fields of biology have found that cellular heterogeneity widely exists in all cell populations.
Analysis in bulk assays may not accurately reflect true biology of individual cells. The studies
in single-cell level help to reveal critical secrets concealed in conventional population-averaged
measurements and provide valuable insights into many unsolved problems in fundamental
biology and human diseases.
To date, the rapid development of many high-precision tools such as high-resolution
microscopes, patch-clamp techniques, micropipettes, optical tweezers etc., has enabled the
single-cell manipulation and exploration. However, those techniques are typically low
throughput, of which the sampling volume is far below statistical level. To extract meaningful
statistics and accurately assess heterogeneities among cell populations, the number of single
cells for analysis should be large enough, which requires high-throughput single-cell analysis
techniques. To date, many novel techniques with greatly enhanced throughput have been
established. Among them, flow cytometry is a very powerful tool pioneering in high-throughput
single-cell studies, which is able to analyze many thousand single cells per second.
In recent years, microfluidic platforms have also been widely utilized for high-throughput
single-cell analyses by leveraging the well-designed structures with sizes matching well with
cells. With optimized geometries, these platforms have many advantages over conventional
methods, such as high throughput, high sensitivity, improved parallelization and low cost.
In the past four years, my studies were basically focused on developing convenient and
effective microchip-based methods for high-throughput single-cell isolation, and combining
different techniques for single-cell quantitative analysis, including fluorescence microscopy
and mass spectrometry. On the basis of developed single-cell isolation technique, I also
explored its applications in single-cell pairing, which exhibited great potentials in the study of
cell-cell interactions and controllable cell fusion.
In the first chapter, an overview of single-cell studies including their significance, current
advances and challenges are present. Then a brief introduction about single-cell pairing is given.
In the rest chapters, I present the research projects completed during my PhD study, including
centrifugation-assisted single-cell trapping (CAScT) in a truncated cone-shaped microwell
array (TCMA) chip for the real-time observation of cellular apoptosis, fast single-cell patterning
for study of drug-induced phenotypic alterations of HeLa cells using time-of-flight secondary
ion mass spectrometry and cell pairing and polyethylene glycol (PEG)-mediated cell fusion
using two-step centrifugation-assisted single-cell trapping.
Post a Comment