Improving image quality in reflection confocal microscopy involving gold nanoparticles and osmotically active immersion liquids

Veksler, B A; Lemelle, Arnaud; Kozhevnikov, Ilya O.; Akchurin, Garif G.; Meglinski, Igor

doi:10.1134/s0030400x11030246

Cited by 3 publications

(2 citation statements)

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“…The perceived centre of the detected region is not perfectly aligned in all cases; this may, in part, relate to the different reflected rays that are collected from irregular shaped NP and agglomerates under the different illumination patterns. Agglomerate shape, size and orientation with respect to the optical axis are all known factors that contribute to the angle of reflectance, and therefore the perceived centre of detection [ 52 – 54 ]. Additionally, problems may arise from the dish not being completely flat during acquisitions.…”

Section: Resultsmentioning

confidence: 99%

“…In other cases, small NP clusters may be masked by larger nearby clusters. The light scattered by an object within a sample is dependent upon physical properties such as size and density [ 53 , 54 ]. It is therefore possible that regions not detected on SIM or RCM, but detected on TEM are not sufficiently agglomerated or aggregated and therefore do not give rise to a detectable signal in the reflectance techniques.…”

Section: Resultsmentioning

confidence: 99%

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Comparison of Confocal and Super-Resolution Reflectance Imaging of Metal Oxide Nanoparticles

et al. 2016

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The potential for human exposure to manufactured nanoparticles (NPs) has increased in recent years, in part through the incorporation of engineered particles into a wide range of commercial goods and medical applications. NP are ideal candidates for use as therapeutic and diagnostic tools within biomedicine, however concern exists regarding their efficacy and safety. Thus, developing techniques for the investigation of NP uptake into cells is critically important. Current intracellular NP investigations rely on the use of either Transmission Electron Microscopy (TEM), which provides ultrahigh resolution, but involves cumbersome sample preparation rendering the technique incompatible with live cell imaging, or fluorescent labelling, which suffers from photobleaching, poor bioconjugation and, often, alteration of NP surface properties. Reflected light imaging provides an alternative non-destructive label free technique well suited, but not limited to, the visualisation of NP uptake within model systems, such as cells. Confocal reflectance microscopy provides optical sectioning and live imaging capabilities, with little sample preparation. However confocal microscopy is diffraction limited, thus the X-Y resolution is restricted to ~250 nm, substantially larger than the <100 nm size of NPs. Techniques such as super-resolution light microscopy overcome this fundamental limitation, providing increased X-Y resolution. The use of Reflectance SIM (R-SIM) for NP imaging has previously only been demonstrated on custom built microscopes, restricting the widespread use and limiting NP investigations. This paper demonstrates the use of a commercial SIM microscope for the acquisition of super-resolution reflectance data with X-Y resolution of 115 nm, a greater than two-fold increase compared to that attainable with RCM. This increase in resolution is advantageous for visualising small closely spaced structures, such as NP clusters, previously unresolvable by RCM. This is advantageous when investigating the subcellular trafficking of NP within fluorescently labelled cellular compartments. NP signal can be observed using RCM, R-SIM and TEM and a direct comparison is presented. Each of these techniques has its own benefits and limitations; RCM and R-SIM provide novel complementary information while the combination of modalities provides a unique opportunity to gain additional information regarding NP uptake. The use of multiple imaging methods therefore greatly enhances the range of NPs that can be studied under label-free conditions.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Comparison of Confocal and Super-Resolution Reflectance Imaging of Metal Oxide Nanoparticles

et al. 2016

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Noninvasive visualization of intraepidermal and subepidermal blisters in vesiculobullous skin disorders by in vivo reflectance confocal microscopy

et al. 2011

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Modern Micro and Nanoparticle-Based Imaging Techniques

Ryvolová

Chomoucká

Drbohlavová

et al. 2012

Sensors

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The requirements for early diagnostics as well as effective treatment of insidious diseases such as cancer constantly increase the pressure on development of efficient and reliable methods for targeted drug/gene delivery as well as imaging of the treatment success/failure. One of the most recent approaches covering both the drug delivery as well as the imaging aspects is benefitting from the unique properties of nanomaterials. Therefore a new field called nanomedicine is attracting continuously growing attention. Nanoparticles, including fluorescent semiconductor nanocrystals (quantum dots) and magnetic nanoparticles, have proven their excellent properties for in vivo imaging techniques in a number of modalities such as magnetic resonance and fluorescence imaging, respectively. In this article, we review the main properties and applications of nanoparticles in various in vitro imaging techniques, including microscopy and/or laser breakdown spectroscopy and in vivo methods such as magnetic resonance imaging and/or fluorescence-based imaging. Moreover the advantages of the drug delivery performed by nanocarriers such as iron oxides, gold, biodegradable polymers, dendrimers, lipid based carriers such as liposomes or micelles are also highlighted.

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Improving image quality in reflection confocal microscopy involving gold nanoparticles and osmotically active immersion liquids

Cited by 3 publications

References 25 publications

Comparison of Confocal and Super-Resolution Reflectance Imaging of Metal Oxide Nanoparticles

Comparison of Confocal and Super-Resolution Reflectance Imaging of Metal Oxide Nanoparticles

Noninvasive visualization of intraepidermal and subepidermal blisters in vesiculobullous skin disorders by in vivo reflectance confocal microscopy

Modern Micro and Nanoparticle-Based Imaging Techniques

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