STED super-resolution imaging of mitochondria labeled with TMRM in living cells

Ishigaki, M; Iketani, Masumi; Sugaya, Maki; Takáhashi, Mayumí; Tanaka, Masashi; Hattori, Satoshi; Ohsawa, Ikuroh

doi:10.1016/j.mito.2016.03.009

Cited by 36 publications

(32 citation statements)

References 49 publications

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“…Cationic rhodamine derivatives localize to mitochondria 35 , but do not blink spontaneously 8 . We synthesized the cationic rhodamine derivative RhoB-TCO (Fig.…”

Section: Resultsmentioning

confidence: 96%

Long time-lapse nanoscopy with spontaneously blinking membrane probes

et al. 2017

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Long time-lapse, diffraction-unlimited super-resolution imaging of cellular structures and organelles in living cells is highly challenging, as it requires dense labeling, bright, highly photostable dyes, and non-toxic conditions. We developed a set of high-density, environment-sensitive (HIDE) membrane probes based on HMSiR that assemble in situ and enable long time-lapse, live cell nanoscopy of discrete cellular structures and organelles with high spatio-temporal resolution. HIDE-enabled nanoscopy movies are up to 50x longer than movies obtained with labeled proteins, reveal the 2D dynamics of the mitochondria, plasma membrane, and filopodia, and the 2D and 3D dynamics of the endoplasmic reticulum in living cells. These new HIDE probes also facilitate the acquisition of live cell, two-color, super-resolution images, greatly expanding the utility of nanoscopy to visualize processes and structures in living cells.

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“…Cationic rhodamine derivatives localize to mitochondria 35 , but do not blink spontaneously 8 . We synthesized the cationic rhodamine derivative RhoB-TCO (Fig.…”

Section: Resultsmentioning

confidence: 96%

Long time-lapse nanoscopy with spontaneously blinking membrane probes

et al. 2017

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“…HT22 cells, derived from primary-cultured mouse hippocampal neurons [32], serve as a cell line of hippocampus neurons of the brain. In this study, we applied the stimulated emission depletion (STED) microscopy technique to study the structure of mitochondria in HT22 cells under TiO 2 NPs insult, since it has the advantage of higher resolution than the conventional confocal microscopy [33,34].…”

Section: Introductionmentioning

confidence: 99%

Titanium Dioxide Nanoparticles Induce Mitochondrial Dynamic Imbalance and Damage in HT22 Cells

Zhao

Chen

Zhong

et al. 2019

Journal of Nanomaterials

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Mitochondria, as dynamic organelles, are precisely regulated by fusion and fission. The dynamic balance of fusion and fission controls mitochondrial morphology and their subcellular location and function. Exposure to titanium dioxide nanoparticles (TiO2 NPs) may cause serious health problems. However, how TiO2 NPs affect the mitochondrial dynamics remains unclear. In the present study, we investigated the changes of mitochondrial dynamics in the TiO2NPs-treated HT22 cells by confocal and stimulated emission depletion (STED) microscopy. The confocal images demonstrated obvious changes in the average length and density of the mitochondria after TiO2 NPs treatment, while STED images further obtained the nanoscale submitochondrial structures of the mitochondria under TiO2 NPs insult. The fluorescence intensity distributions suggested that mitochondria fragmented in the TiO2 NPs-treated cells. TiO2 NPs treatment caused mitochondrial dynamic imbalance due to the imbalanced expression of dynamin-related protein 1 (Drp1) and optic atrophy 1 (Opa1). Furthermore, we examined the levels of oxidative stress and mitochondrial membrane potential (MMP) and the generation of adenosine triphosphate (ATP), which revealed the damage of mitochondria under TiO2 NPs exposure. Meanwhile, the significant changes of expressions of B-cell lymphoma 2-associated X protein (Bax), B-cell lymphoma 2 (Bcl-2), cytochrome c (Cyt C), and caspase 9 demonstrated that TiO2 NPs treatment activated the mitochondrial-related apoptosis pathway. These cellular events can be largely prevented via cell incubation with mitoTEMPO, a mitochondria-targeted superoxide scavenger. Our results confirm that TiO2 NPs targeted the mitochondria, inducing mitochondrial dynamic imbalance and damage in HT22 cells. Our study provides an insightful understanding of the mechanisms underlying TiO2 NPs cytotoxicity.

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“…Recently, Xi et al narrowed the axially PSF of STED by employing the interference between the PSF and its re°ection from a mirror placed behind the specimen. 27 Despite its short time from the invention, STED microscopy has made many contributions to the observation of subcellular structures such as endoplasmic reticulum, 28 mitochondria 29,30 and cytoskeletons. 31,32 Because of complicated components involved in signal transduction between cells, dualcolor STED [33][34][35] and even multi-color STED 36,37 have been established to study several active complexes at the same time.…”

Section: Stimulated Emission Depletion (Sted) Microscopymentioning

confidence: 99%

Super-resolution microscopy and its applications in neuroscience

Wang

Zhu

et al. 2017

J. Innov. Opt. Health Sci.

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Optical microscopy promises researchers to see most tiny substances directly. However, the resolution of conventional microscopy is restricted by the di®raction limit. This makes it a challenge to observe subcellular processes happened in nanoscale. The development of superresolution microscopy provides a solution to this challenge. Here, we brie°y review several commonly used super-resolution techniques, explicating their basic principles and applications in biological science, especially in neuroscience. In addition, characteristics and limitations of each technique are compared to provide a guidance for biologists to choose the most suitable tool.

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STED super-resolution imaging of mitochondria labeled with TMRM in living cells

Cited by 36 publications

References 49 publications

Long time-lapse nanoscopy with spontaneously blinking membrane probes

Long time-lapse nanoscopy with spontaneously blinking membrane probes

Titanium Dioxide Nanoparticles Induce Mitochondrial Dynamic Imbalance and Damage in HT22 Cells

Super-resolution microscopy and its applications in neuroscience

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