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Subcellular tracking and quantification of metals was achieved through use of the x-ray fluorescence microprobe (XFM) of beam-line 2-ID-E of the Advanced Photon Source.
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We demonstrate its superior performance by resolving subcellular structures and tracking single-mitochondrion dynamic processes in live model organisms.
For organelles that contain their own DNA, such as mitochondria and chloroplasts, researchers can follow the organelle purification with a DNA-isolation step to track subcellular genetics.
In the first, we linked YFP to the C terminus of HKI or HKII (HKI-YFP and HKII-YFP) to track subcellular location.
Fluorescence labeling and tracking of subcellular organelles and proteins are considered as a powerful tool to reveal the mystery of cellular activities.
Several of these applications have been hinted at in the text (for example semi-automated segmentation, extraction of 3D data from complex cell shapes (like lobed cells), tracking of subcellular elements).
Our results using live cell imaging to track the subcellular distributions of HKI and HKII provide direct support for this hypothesis.
In the present study, we have expressed HKI and HKII tagged with YFP in Chinese Hamster Ovary (CHO) cells to track their subcellular location in real time and their mobilization in response to substrates.
The possibility to control the fluorescence properties enabled numerous innovative applications, for instance, regional optical marking and tracking of cells, subcellular organelles or proteins [3], [15], [21].
Larvae were processed for histological and ultrastructural analyses to detect the main affected organs, to look for specific lesions at the subcellular level and to image and track CNPs into tissues.
The technique uses novel fluorescence nuclear track detectors with fluorescence confocal laser scanning microscopy in the beam line to visualize particle track traversals within the subcellular compartments of live cells within seconds after injury.
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