Abstract: A method for measuring autophagosome flux is provided. The autophagosome pool size in a single cell is quantified, where the pool size is the total number of autophagosomes in the cell. Fusion between the autophagosomes and lysosomes in the cell is then inhibited. The autophagosome pool size is quantified over one or more time points after fusion has been inhibited, and the autophagosome flux is calculated as the initial rate of change of the autophagosome pool size at the time point after fusion has been inhibited. The method can be used to determine basal autophagosome flux, whether a cell is diseased or dysfunctional, or to diagnose a subject with a disease, disorder or dysfunction. The transition time, the time required to clear an autophagosome pool, can also be derived from the autophagosome flux. A molecule can also be characterized according to its ability to modulate autophagosome flux in a cell.

Abstract: A computer-implemented method and a system are provided for visualising colocalised fluorescence signals. The method accesses signal intensity data obtained from a first fluorescence channel and a second fluorescence channel in which the signal intensity data is associated with voxels in an image. A regression factor on the signal intensity data is calculated to generate a regression parameter corresponding to a degree of correlation between the signal intensity data obtained from the first and second fluorescence channels The signal intensity data is mapped to the regression parameter and colourmap values are assigned to each voxel based on the mapped signal intensity data in which colourmap values of voxels embodying poorly correlated signal intensity data are reduced. The method renders the voxels in the image in colours according to their colourmap values to visualise colocalisation in the image.

Abstract: A method, device and system for determining autophagic flux are claimed. The levels of proteins which change with increased or decreased autophagy are determined in a sample. The change in the level of each protein is quantified in order to obtain the autophagic flux. This can be compared to a sample flux range associated with autophagy dysfunction or ageing patterns. Diseases or conditions which may be diagnosed include neurodegenerative conditions such as Alzheimer's disease and dementia, cancer, heart conditions, immune conditions or aging-related conditions. The device for determining autophagic flux comprises a housing, receiving zones configured for receiving a substrate and a biological sample, and a set of electrodes for each receiving zone. The device is connectable to circuitry that determines an electrical property of each substrate and uses this to determine the autophagic flux.

Abstract: A method of determining the location and quantity of mitochondrial fission, fusion and depolarisation events that occur in a cell is provided. Using a three-dimensional time lapse image sequence of a cell, the method identifies which of the mitochondria in a cell had depolarised or undergone fission or fusion in the interval between the acquisition of the earlier and later images, indicates the locations of the fission, fusion and depolarisation events, and generates a count of the number of mitochondrial fission, fusion and/or depolarisation events. The method can be used to diagnose a disease or condition associated with mitochondrial dysfunction, such as neurodegenerative disease, cancer or ischaemic heart disease. The method can further be used to screen a compound or composition for use in preventing or treating a disease or condition associated with mitochondrial dysfunction. The method can be computer-implemented, and a computer program product is provided.

Abstract: A computer-implemented method and a system are provided for visualising colocalised fluorescence signals. The method accesses signal intensity data obtained from a first fluorescence channel and a second fluorescence channel in which the signal intensity data is associated with voxels in an image. A regression factor on the signal intensity data is calculated to generate a regression parameter corresponding to a degree of correlation between the signal intensity data obtained from the first and second fluorescence channels The signal intensity data is mapped to the regression parameter and colourmap values are assigned to each voxel based on the mapped signal intensity data in which colourmap values of voxels embodying poorly correlated signal intensity data are reduced. The method renders the voxels in the image in colours according to their colourmap values to visualise colocalisation in the image.

Abstract: A method, device and system for determining autophagic flux are claimed. The levels of proteins which change with increased or decreased autophagy are determined in a sample. The change in the level of each protein is quantified in order to obtain the autophagic flux. This can be compared to a sample flux range associated with autophagy dysfunction or ageing patterns. Diseases M or conditions which may be diagnosed include neurodegenerative conditions such as Alzheimer's disease and dementia, cancer, heart conditions, immune conditions or aging-related conditions. The device for determining autophagic flux comprises a housing, receiving zones configured for receiving a substrate and a biological sample, and a set of electrodes for each receiving zone. The device is connectable to circuitry that determines an electrical property of each substrate and uses this to determine the autophagic flux.

Abstract: A method for measuring autophagosome flux is provided. The autophagosome pool size in a single cell is quantified, where the pool size is the total number of autophagosomes in the cell. Fusion between the autophagosomes and lysosomes in the cell is then inhibited. The autophagosome pool size is quantified over one or more time points after fusion has been inhibited, and the autophagosome flux is calculated as the initial rate of change of the autophagosome pool size at the time point after fusion has been inhibited. The method can be used to determine basal autophagosome flux, whether a cell is diseased or dysfunctional, or to diagnose a subject with a disease, disorder or dysfunction. The transition time, the time required to clear an autophagosome pool, can also be derived from the autophagosome flux. A molecule can also be characterized according to its ability to modulate autophagosome flux in a cell.