This experiment aims to capture the dynamic process of phagosome maturation and phagolysosome biogenesis in phagocytes. By utilizing fluorescent conjugated dex strands, researchers can visualize lysosomal compartments and analyze the transfer of lysosomal material to phagosomes.
The overall goal of the following experiment is to capture the real time dynamic process of phagosome maturation and phagolysosome biogenesis in phagocytes. This is achieved by loading the lysosomal compartments of the cells with fluorescent conjugated dex strand to enable the visualization of lysosomal compartments and transfer of its luminal cargo to phagosomes. As a second step, microparticles are added, which act as a phagocytic model.
Next, the delivery of lysosomal material to the phagosomes is analyzed using live sale imaging and subsequent digital image processing. In order to quantify the phagosome maturation process, the results show the effect of various factors on the process of phagosome maturation based on the delivery of lysosomal content to the phagosomes. The main advantage of this technique of existing methods, such as the evaluation of macrophage maturation in fixed cells is the quantitative readout with a high temporal resolution Begin by dissolving Texas red labeled 70 kilodalton DExT strand or Dex 70 KD in PBS at 20 milligrams per milliliter and store aliquots at minus 20 degrees Celsius.
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