Using a Single Diamond NV Center for Nanoscale Fluorescence Lifetime Imaging

Beams, Ryan; Smith, Dallas; Johnson, Timothy W.; Oh, Sang Hyun; Novotny, Lukas; Vamivakas, A. Nick

doi:10.1364/cqo.2013.m6.55

Cited by 2 publications

(1 citation statement)

References 7 publications

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“…In spite of many difficulties, fluorescence lifetime imaging (FLIM) has been very successful and lead to many practical applications [23–27]. Further, the development of pulsed lasers and laser diodes that are now widely available with typical imaging systems opens many new applications for FLIM.…”

Section: Introductionmentioning

confidence: 99%

Multiple-pulse pumping for enhanced fluorescence detection and molecular imaging in tissue

Rich

Gryczyński

Fudala

et al. 2014

Methods

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Applications of fluorescence based imaging techniques for detection in cellular and tissue environments are severely limited by autofluorescence of endogenous components of cells, tissue, and the fixatives used in sample processing. To achieve sufficient signal-to-background ratio, a high concentration of the probe needs to be used which is not always feasible. Since typically autofluorescence is in the nanosecond range, long-lived fluorescence probes in combination with time-gated detection can be used for suppression of unwanted autofluorescence. Unfortunately, this requires the sacrifice of the large portion the probe signal in order to sufficiently filter the background. We report a simple and practical approach to achieve a many-fold increase in the intensity of a long-lived probe without increasing the background fluorescence. Using controllable, well separated bursts of closely spaced laser excitation pulses, we are able to highly increase the fluorescence signal of a long-lived marker over the endogenous fluorescent background and scattering, thereby greatly increasing detection sensitivity. Using a commercially available confocal microscopy system equipped with a laser diode and time correlated single photon counting (TCSPC) detection, we are able to enhance the signal of a long-lived Ruthenium (Ru)-based probe by nearly an order of magnitude. We used 80 MHz bursts of pulses (12.5 ns pulse separation) repeated with a 320 kHz repetition rate as needed to adequately image a dye with a 380 ns lifetime. Just using 10 pulses in the burst increases the Ru signal almost 10 fold without any increase in the background signal.

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Section: Introductionmentioning

confidence: 99%

Multiple-pulse pumping for enhanced fluorescence detection and molecular imaging in tissue

Rich

Gryczyński

Fudala

et al. 2014

Methods

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Functionalized Nanodiamonds for Targeted Neuronal Electromagnetic Signal Detection

Costa,

Camarneiro,

Marote

et al. 2024

ACS Appl. Mater. Interfaces

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Intracellular sensing technologies necessitate a delicate balance of spatial resolution, sensitivity, biocompatibility, and stability. While existing methods partially fulfill these criteria, none offer a comprehensive solution. Nanodiamonds (NDs) harboring nitrogenvacancy (NV) centers have emerged as promising candidates due to their sensing capabilities under biological conditions and their ability to meet all aforementioned requirements. This study focuses on expanding the application of NDs and NV center-based sensing to neuronal contexts by investigating their functionalization and subsequent effects on three distinct cell lines relevant to neurodegenerative disease research. Our study concentrates on positioning fluorescent NDs (FNDs) with NV center point defects onto neuronal cell surfaces. Achieving this through specific antibody attachment enhances the proximity of FND to neurites, facilitating the detection of local action potentials. Targeting voltage-dependent calcium channels (Cav2.2) with biotin−streptavidin-bound antibodies enables the precise positioning of FNDs. The functionalized FNDs (f-FNDs) show increased size and zeta potential, confirming the antibody presence without compromising cell viability. Two-color confocal imaging and co-localization algorithms are employed to further attest to the success of the functionalization. The f-FNDs are applied to cell cultures of three cell lines: SH-SY5Y, differentiated dopaminergic neurons, and hippocampal rat neurons; their biocompatibility and effects on synaptic activity are explored. Moreover, preliminary total internal reflection fluorescence -optically detected magnetic resonance (TIRF-ODMR) experiments across cellular sites demonstrate the magnetic field sensitivity of our sensor network. The successful establishment of this sensor network provides a platform for characterizing neuronal signaling in healthy models and conditions mimicking Parkinson's disease.

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Using a Single Diamond NV Center for Nanoscale Fluorescence Lifetime Imaging

Abstract: We study the near-field optical properties of a diamond nanocrystal hosting a single NV center. Using a single NV center as a probe of the electromagnetic mode structure, we demonstrate nanoscale fluorescence lifetime imaging microscopy.

Cited by 2 publications

References 7 publications

Multiple-pulse pumping for enhanced fluorescence detection and molecular imaging in tissue

Multiple-pulse pumping for enhanced fluorescence detection and molecular imaging in tissue

Functionalized Nanodiamonds for Targeted Neuronal Electromagnetic Signal Detection

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