Effects on immune cell viability, morphology and proliferation in a sub-microliter cell sampler system
Independent thesis Advanced level (degree of Master (Two Years)), 20 credits / 30 HE creditsStudent thesis
Today, most traditional method used in the research of immune cells, such as flow cytometry and microscopy, are based on average values of cell responses. However, immune cells are heterogeneous and respond differently to a given stimuli. There is also a risk that important, but rare, behaviors of individual cells are missed when a larger population of immune cells is analyzed. Also, flow cytometry and microscopy do not allow long-term survival of cells; these methods lack the ability to do dynamic long-term analysis of motile immune cells, i.e. studies of cell-cell interactions, morphology and proliferation.
In a patient who is affected by cancer, the cell heterogeneity contributes to the ability to battle various types of cancer or virus infections. In an outbreak, immune cells recognize and kill tumor cells. However, the number of specific immune cells is sometimes too few to kill all the tumor cells in a successful way. One way to help these patients is to isolate, select out and cultivate the active immune cells with capacity to kill tumor cells.
The Cell Physic Laboratory (a part of the department of Applied Physics) at the Royal Institute of Technology (KTH) has developed a method for single-cell analysis where the immune cells are trapped in microwells in a silicon chip. The immune cells are then studied by using fluorescence microscopy in an inverted setup. The method enables high-throughput experiments due to the parallelization. Furthermore, since the immune cells survive long periods in the chip, the cells can be analyzed over several days up to weeks. The research group has also developed a semi-automatic ‘cell-picker’. The cell-picker will be used in combination with the developed method for single-cell analysis, which enables picking of cells of interest.
In this report, experiments for the characterization and evaluation of the biocompatibility of two generations of the cell-picker will be presented. The experiments include development of a protocol for the cell-picking process, studies of the survival time of transferred cells for both generation of the cell-picker and studies of surface coating in the chip in order to increase the biocompatibility. The preliminary results indicate that the cell-picker has potential to be used as a selection tool for immune cells of interest.
Place, publisher, year, edition, pages
2013. , 71 p.
Cell-picker, sub-microliter cell sampler system, immune cells, T cells, NK cells, B cells, cell viability, morphology, proliferation, surface coating, microchip, wells, silicon chip, single-cell analysis, flow system, fluorescence microscopy, cell staining, characterization
Engineering and Technology
IdentifiersURN: urn:nbn:se:liu:diva-89982ISRN: LITH-IFM-A-EX--13/2714--SEOAI: oai:DiVA.org:liu-89982DiVA: diva2:610689
Kungliga Tekniska Högskolan (KTH)
Subject / course
UppsokPhysics, Chemistry, Mathematics