Journal
BULLETIN OF THE KOREAN CHEMICAL SOCIETY
Volume 36, Issue 11, Pages 2589-2595Publisher
WILEY-V C H VERLAG GMBH
DOI: 10.1002/bkcs.10521
Keywords
dSTORM; High temporal resolution; Intracellular Mg2+; Single-cell analysis; Super-resolution imaging
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Funding
- Basic Science Research Program through the National Research Foundation of Korea (NRF) - Ministry of Education, Science and Technology [2015R1A2A2A01003839]
- National Research Foundation of Korea [2015R1A2A2A01003839] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)
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Magnesium ion (Mg2+) is an important intracellular component that plays a fundamental role in many cellular processes. The super-localization of intracellular Mg2+ signaling and wave propagation in a single cell with subdiffraction-limit image resolution is demonstrated for the first time by direct stochastic optical reconstruction microscopy (dSTORM). Mag-fluo-4-AM, an intracellular Mg2+ fluorescent indicator dye, is used for the fluorescence imaging of intracellular Mg2+ in human embryonic kidney 293 (HEK-293) cells as a model. Mag-fluo-4-AM is photoswitched between its fluorescent and dark states and the resulting super-resolution cell image reveals uniform and specific labeling of Mg2+ with thousands of fluorophores per cell and localization precision of 21 +/- 1nm. Furthermore, the real-time dynamics of Mg2+ wave propagation are studied by dSTORM. The sparks and waves show random temporal propagation patterns in nonhomogeneous substructures with 0.2s time intervals. Thus, dSTORM allows characterization of intracellular processes at sub-nanometre spatial resolution with high temporal resolution.
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