This PhD thesis is concerned with developing a methodology for early diagnosis of cancer by comparing the resonant frequencies in the amplitude spectra obtained during a vibration test using the AFM or, by comparing the stiffness properties of single cancerous and normal cells obtained using a resonant technique. As there is no reliable data in the literature to prove the existence of resonant frequencies of single cells, this work pioneers the search for resonant frequencies of related microspherical soft bodies using the AFM. Experiments to investigate the resonant behaviour of single cells depends on various parameters which are difficult to control; for example, the cell type, deciding at what stage the cell should be tested during the culturing process, determining the nucleus size, determining the cytoskeleton integrity and designing an appropriate vibration test setup among others. For this reason, agarose microspheres were selected to carry out preliminary work as these samples have similar properties to human cells and their resonances are affected by fewer variables. Although these micrometric spheres were tested under different conditions, no clear resonant behaviour was found at frequencies below 20 kHz and, only wide curves (interpreted as highly damped peaks of resonance) in the interval ranging from 20 kHz to 100 kHz were observed. By considering those curves as the quadrupole (Qp) vibration mode, approximate stiffness values for the agarose microspheres were found to be in between 37 kPa and 72 kPa. These values are similar to those obtained during an indentation test performed on the same samples whic¬¬h gave Young’s modulus values ranging from 10 kPa to 200 kPa. In order to gain a greater insight into the vibration test performed on microscopic samples, the research was extended to include agarose spheres of millimetric size. The characterization of these samples was carried out using an innovative purpose-built experimental setup. For the vibration test, a PZT based excitation device and a vibro-acoustic sensor were designed and constructed. The amplitude spectra of the vibration tests performed on millimetric samples consistently showed at least three peaks of resonance from which after the numerical simulation of the vibration test were interpreted as the quadrupole (Qp) and octupole (Op) vibration modes. Using this information, stiffness values for the samples ranging from 100 kPa to 700 kPa were calculated. In order to obtain the stiffness of the millimetric samples using a different technique, an experimental setup was constructed to perform a compression test. However, due to high viscoleasticity of the samples, it was not possible to obtain a standard compression curve necessary for their mechanical characterization. The results obtained from the tests on millimetric agarose samples demonstrate that spheres made of this material are able to provide measurable vibrational characteristics. Consequently, this methodology can be further implemented on micrometric samples and possibly on human cells to detect their resonant frequencies and equivalent stiffness values which can be used as a cancer marker. From the vibrational experiments on millimetric samples, it was noticed that the excitation mechanism plays an important role and for this reason future work is proposed to continue in this direction.
| Identifer | oai:union.ndltd.org:bl.uk/oai:ethos.bl.uk:626933 |
| Date | January 2014 |
| Creators | Yescas, Jorge Arturo |
| Publisher | University of Manchester |
| Source Sets | Ethos UK |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
| Source | https://www.research.manchester.ac.uk/portal/en/theses/vibrational-study-of-agarose-spheres-of-millimetric-and-micrometric-size(c4bf7877-f6cf-48b8-bed2-ece48e230c5d).html |
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