Parkinson’s disease (PD) is characterized by progressive dopaminergic cell loss in the substantia nigra (SN) and elevated iron levels demonstrated by autopsy. Direct visualization of iron with live imaging techniques has not yet been successful. The aim of this study is to visualize and quantify the distribution of cellular iron using an intrinsic iron hyperspectral fluorescence signal. The 1-methyl-4-phenylpyridinium (MPP+)-induced cellular model of PD was established in SHSY5Y cells exposed to iron with ferric ammonium citrate (FAC, 100 μM). The hyperspectral fluorescence signal of iron was examined using a high-resolution dark-field optical microscope system with signal absorption for the visible/near infrared spectral range. The 6-h group showed heavy cellular iron deposition compared with the 1-h group. The cellular iron was dispersed in a small particulate form, whereas the extracellular iron was aggregated. In addition, iron particles were found to be concentrated on the cell membrane/edge of shrunken cells. The iron accumulation readily occurred in MPP+-induced cells, which is consistent with previous studies demonstrating elevated iron levels in the SN. This direct iron imaging could be applied to analyze the physiological role of iron, and its application might be expanded to various neurological disorders involving metals, such as copper, manganese, or zinc.
We report the application of an optical microscope equipped with a high-resolution dark-field condenser for detecting dynamic responses of cellular nanostructures in real time. Our system provides an easy-to-use technique to visualize biological specimens without any staining. This system can visualize the dynamic behavior of nanospheres and nanofibers, such as F-actin, at the leading edges of adjacent neuronal cells. We confirmed that the nanofibers imaged with this high-resolution optical microscopic technique are F-actin by using fluorescence microscopy after immunostaining the F-actin of fixed cells. Furthermore, cellular dynamics are enhanced by applying noncontact electric field stimulation through a transparent graphene electric field stimulator. High-resolution label-free optical microscopy enables the visualization of nanofiber dynamics initiated by filopodial nanofiber contacts. In conclusion, our optical microscopy system allows the visualization of nanoscale cellular dynamics under various external stimuli in real time without specific staining.
Electrical stimulation affects cellular behaviors including division, migration and wound healing [1-3]. Cellular
injury often occurs due to the imbalance of the endogenous electric field [3]. In order to recover from the injury,
wound healing process requires various cellular changes such as regeneration, migration, and the enhancement of cytoskeletal proteins and growth factors. In previous reports, a weak non-contact electric field stimulation (nEFS) accelerates the cell migration as well as cell-to-cell coupling between neuronal cell junction which are accompanied by increasing of cytoskeletal proteins [4, 5]. In this paper, we further investigated the wound
healing effect of the nEFS in the neuronal cells (SHSY5Y cells) with live cell optical imaging. Cells were
cultured over the optically transparent graphenen EF stimulator. Cellular behavioral changes upon nEFS were
recorded with live optical imaging during stimulation of 120 minutes. The ability of wound healing was
significantly enhanced with the nEFS. In particular, nEFS significantly shorten the duration of wound healing
process. Moreover, after treating cells with cytochalasin D, a block polymerization of the actin filaments, the
nEFS significantly enhanced wound healing process of cytochalasin D treated neural cells as compared to the control neural cells. This study suggests that nEFS may provide an effective way to control neural cells repairing
process from cellular injury. Further mechanism study about the effect of nEFS on the wound healing may shed
new light on cellular behavior.
Parkinson’s disease (PD) is characterized by progressive dopaminergic cell loss in the substantianigra (SN)
and elevated iron levels demonstrated by autopsy and with 7-Tesla magnetic resonance imaging. Direct
visualization of iron with live imaging techniques has not yet been successful. The aim of this study is to visualize and quantify the distribution of cellular iron using an intrinsic iron hyperspectral fluorescence
signal. The 1-methyl-4-phenylpyridinium (MPP+)-induced cellular model of PD was established in SHSY5Y cells. The cells were exposed to iron by treatment with ferric ammonium citrate (FAC, 100 μM)
for up to 6 hours. The hyperspectral fluorescence imaging signal of iron was examined usinga high-
resolution dark-field optical microscope system with signal absorption for the visible/ near infrared
(VNIR) spectral range. The 6-hour group showed heavy cellular iron deposition compared with the small
amount of iron accumulation in the 1-hour group. The cellular iron was dispersed in a small, particulate
form, whereas extracellular iron was detected in an aggregated form. In addition, iron particles were
found to be concentrated on the cell membrane/edge of shrunken cells. The cellular iron accumulation readily occurred in MPP+-induced cells, which is consistent with previous studies demonstrating elevated iron levels in the SN in PD. This direct iron imaging methodology could be applied to analyze the
physiological role of iron in PD, and its application might be expanded to various neurological disorders involving other metals, such as copper, manganese or zinc.
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