Thesis Flow Cytometry

Thesis Flow Cytometry-66
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Specific cell populations can be selected for analysis based on combined variables such as fluorescent intensity, cell shape, cell size and texture.

In this webcast the value of imaging flow cytometry for the assessment of acute and chronic leukaemias will be demonstrated.

The imaging functionality allows the localisation of cellular antigens to be determined (i.e.

cell surface, cytoplasm, nuclear), a feature that cannot be achieved by standard flow cytometry.

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Your access to the NCBI website at gov has been temporarily blocked due to a possible misuse/abuse situation involving your site.Imaging flow cytometry has opened new opportunities in the assessment of leukaemia whereby this information can be generated on one automated instrument: an imaging flow cytometer.Image-based flow cytometry combines high-resolution digital images with the quantitative fluorescence information gained from a standard flow cytometer.In the field of imaging flow cytometry, accuracy, simplicity and processing time of image processing algorithms plays a significant role.The microscopic image data acquired by video camera is of low contrast and low resolution. Immunofluorescence experiments were also performed to label cell nuclei isolated from wild type mouse CNS for FANS analysis. When compared to an existing technique, translating ribosome affinity purification, it was found that FANS was able to detect differentially expressed genes between two types of medium spiny neurons in the striatum with similar or higher sensitivity, using transgenic mice with fluorescence proteins labeling the desired cell types.It was found that our staining method successfully labeled neuronal nuclei using a Neu N antibody, but did not label enriched markers of medium spiny neurons.Further studies are needed to increase the signal-to-back-ground ratio of these stainings, which would allow FANS to be applied to wild type animals.We will look at the detection of disrupted PML protein in acute promyelocytic leukaemia and the ability to distinguish between cytoplasmic and nuclear NPM in acute myeloid leukaemia.The webinar will also present exciting new data demonstrating the ability to detect chromosomal abnormalities by FISH in phenotyped leukaemia cells in chronic lymphocytic leukaemia.

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