Nanoparticle discrimination based on wavelength and lifetime-multiplexed cathodoluminescence microscopy

Garming, Mathijs W. H.; Weppelman, I.G.C.; Boer, Pascal de; Martínez, Felipe Perona; Schirhagl, Romana; Hoogenboom, Jacob P.; Moerland, Robert J.

doi:10.1039/c7nr00927e

Cited by 16 publications

(20 citation statements)

References 59 publications

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“…Recently, an analog of lifetime imaging in cathodoluminescence has been demonstrated. In this imaging modality, diamonds have shorter cathodoluminescence lifetimes than the surrounding cell material …”

Section: Labelingmentioning

confidence: 99%

Nanodiamonds and Their Applications in Cells

Chipaux

Laan

Hemelaar

et al. 2018

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Diamonds owe their fame to a unique set of outstanding properties. They combine a high refractive index, hardness, great stability and inertness, and low electrical but high thermal conductivity. Diamond defects have recently attracted a lot of attention. Given this unique list of properties, it is not surprising that diamond nanoparticles are utilized for numerous applications. Due to their hardness, they are routinely used as abrasives. Their small and uniform size qualifies them as attractive carriers for drug delivery. The stable fluorescence of diamond defects allows their use as stable single photon sources or biolabels. The magnetic properties of the defects make them stable spin qubits in quantum information. This property also allows their use as a sensor for temperature, magnetic fields, electric fields, or strain. This Review focuses on applications in cells. Different diamond materials and the special requirements for the respective applications are discussed. Methods to chemically modify the surface of diamonds and the different hurdles one has to overcome when working with cells, such as entering the cells and biocompatibility, are described. Finally, the recent developments and applications in labeling, sensing, drug delivery, theranostics, antibiotics, and tissue engineering are critically discussed.

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Section: Labelingmentioning

confidence: 99%

Nanodiamonds and Their Applications in Cells

Chipaux

Laan

Hemelaar

et al. 2018

Small

Self Cite

185

195

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“…The detected particle-specific wavelength emissions can then discriminate targets, four distinct CL spectra simultaneously have been demonstrated (Glenn et al 2012 ; Morrison et al 2015 ; Niioka et al 2014 ; Tizei and Kociak 2012 ). CL lifetime can be detected as an alternative or additional means to discriminate nanoparticle emissions (Garming et al 2017 ). However, the current limitation of CL towards nanoscale resolution lies in the size of the nanoparticles, most of which are still way beyond the scale of target proteins (Fukushima et al 2014 ; Hemelaar et al 2017 ).…”

Section: Cathodoluminescence (Cl)mentioning

confidence: 99%

ColorEM: analytical electron microscopy for element-guided identification and imaging of the building blocks of life

Pirozzi

Hoogenboom

Giepmans

2018

Histochem Cell Biol

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Nanometer-scale identification of multiple targets is crucial to understand how biomolecules regulate life. Markers, or probes, of specific biomolecules help to visualize and to identify. Electron microscopy (EM), the highest resolution imaging modality, provides ultrastructural information where several subcellular structures can be readily identified. For precise tagging of (macro)molecules, electron-dense probes, distinguishable in gray-scale EM, are being used. However, practically these genetically-encoded or immune-targeted probes are limited to three targets. In correlated microscopy, fluorescent signals are overlaid on the EM image, but typically without the nanometer-scale resolution and limited to visualization of few targets. Recently, analytical methods have become more sensitive, which has led to a renewed interest to explore these for imaging of elements and molecules in cells and tissues in EM. Here, we present the current state of nanoscale imaging of cells and tissues using energy dispersive X-ray analysis (EDX), electron energy loss spectroscopy (EELS), cathodoluminescence (CL), and touch upon secondary ion mass spectroscopy at the nanoscale (NanoSIMS). ColorEM is the term encompassing these analytical techniques the results of which are then displayed as false-color at the EM scale. We highlight how ColorEM will become a strong analytical nano-imaging tool in life science microscopy.

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“…Robust luminescent markers that can withstand electron beam exposure offer great perspective in correlative cathode luminescence (CL) and scanning electron microscopy (SEM) experiments. Correlative CL-SEM studies have been performed previously on Y 2 O 3 :Ln 3+ NPs of ß70 nm (Fukushima et al, 2016) and on ß37 nm LuAG:Ce 3+ NPs (Glenn et al, 2012;Garming et al, 2017). However, cathode luminescence measurements on LuAG:Ce 3+ NPs have revealed a fluctuation in intensity in time, possibly due to bleaching or drifting of the sample during electron exposure (Garming et al, 2017).…”

Section: Introductionmentioning

confidence: 99%

“…Correlative CL-SEM studies have been performed previously on Y 2 O 3 :Ln 3+ NPs of ß70 nm (Fukushima et al, 2016) and on ß37 nm LuAG:Ce 3+ NPs (Glenn et al, 2012;Garming et al, 2017). However, cathode luminescence measurements on LuAG:Ce 3+ NPs have revealed a fluctuation in intensity in time, possibly due to bleaching or drifting of the sample during electron exposure (Garming et al, 2017). Robust particles that exhibit CL are also of interest in CL-STEM (Kociak et al, 2017) and integrated CLEM systems that allow CL detection (Haring et al, 2017).…”

Section: Introductionmentioning

confidence: 99%

Towards robust and versatile single nanoparticle fiducial markers for correlative light and electron microscopy

Hest

Agronskaia

Fokkema

et al. 2019

Journal of Microscopy

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SummaryFiducial markers are used in correlated light and electron microscopy (CLEM) to enable accurate overlaying of fluorescence and electron microscopy images. Currently used fiducial markers, e.g. dye‐labelled nanoparticles and quantum dots, suffer from irreversible quenching of the luminescence after electron beam exposure. This limits their use in CLEM, since samples have to be studied with light microscopy before the sample can be studied with electron microscopy. Robust fiducial markers, i.e. luminescent labels that can (partially) withstand electron bombardment, are interesting because of the recent development of integrated CLEM microscopes. In addition, nonintegrated CLEM setups may benefit from such fiducial markers. Such markers would allow switching back from EM to LM and are not available yet.Here, we investigate the robustness of various luminescent nanoparticles (NPs) that have good contrast in electron microscopy; 130 nm gold‐core rhodamine B‐labelled silica particles, 15 nm CdSe/CdS/ZnS core–shell–shell quantum dots (QDs) and 230 nm Y2O3:Eu3+ particles. Robustness is studied by measuring the luminescence of (single) NPs after various cycles of electron beam exposure. The gold‐core rhodamine B‐labelled silica NPs and QDs are quenched after a single exposure to 60 ke− nm–2 with an energy of 120 keV, while Y2O3:Eu3+ NPs are robust and still show luminescence after five doses of 60 ke− nm–2. In addition, the luminescence intensity of Y2O3:Eu3+ NPs is investigated as function of electron dose for various electron fluxes. The luminescence intensity initially drops to a constant value well above the single particle detection limit. The intensity loss does not depend on the electron flux, but on the total electron dose. The results indicate that Y2O3:Eu3+ NPs are promising as robust fiducial marker in CLEM.Lay DescriptionLuminescent particles are used as fiducial markers in correlative light and electron microscopy (CLEM) to enable accurate overlaying of fluorescence and electron microscopy images. The currently used fiducial markers, e.g. dyes and quantum dots, loose their luminescence after exposure to the electron beam of the electron microscope. This limits their use in CLEM, since samples have to be studied with light microscopy before the sample can be studied with electron microscopy. Robust fiducial markers, i.e. luminescent labels that can withstand electron exposure, are interesting because of recent developments in integrated CLEM microscopes. Also nonintegrated CLEM setups may benefit from such fiducial markers. Such markers would allow for switching back to fluorescence imaging after the recording of electron microscopy imaging and are not available yet.Here, we investigate the robustness of various luminescent nanoparticles (NPs) that have good contrast in electron microscopy; dye‐labelled silica particles, quantum dots and lanthanide‐doped inorganic particles. Robustness is studied by measuring the luminescence of (single) NPs after various cycles of electron beam exposure. The dye‐labelle...

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Nanoparticle discrimination based on wavelength and lifetime-multiplexed cathodoluminescence microscopy

Cited by 16 publications

References 59 publications

Nanodiamonds and Their Applications in Cells

Nanodiamonds and Their Applications in Cells

ColorEM: analytical electron microscopy for element-guided identification and imaging of the building blocks of life

Towards robust and versatile single nanoparticle fiducial markers for correlative light and electron microscopy

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