Detection of triple antibody-binding lymphocytes in standard single laser flow cytometry using colloidal gold, fluorescein and phycoerythrin as labels

Festin, Roger; Björklund, Birgitta; Tötterman, Thomas H.

doi:10.1016/0022-1759(87)90211-0

Cited by 28 publications

(16 citation statements)

References 12 publications

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“…Flow cytometry (FCM) has been combined with both DNA stains and monoclonal antibody labels for the examination of leucocytes. Multi-parameter immunofluorescence FCM techniques are well established for the identification of lymphocyte sub-populations (Lanier and Loken 1984;Parks et al 1984;Festin et al 1987)) including the discrimination of tumorous from healthy lymphocytes (Begg and Hofland 1991). For multi-parameter immunofluorescence FCM, either single or dual laser systems may be used.…”

Section: Introductionmentioning

confidence: 99%

Detection of low levels of specific Salmonella species by fluorescent antibodies and flow cytometry

McClelland

Pinder

1994

Journal of Applied Bacteriology

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The use of fluorescently-labelled monoclonal antibodies, with detection by multi-parameter flow cytometry, was investigated for the rapid detection of salmonellas in pure cultures. Accurate detection of specific Salmonella serotypes was demonstrated down to levels of below 10(4) cells ml-1 (within 30 min) and 1 cell ml-1 (after 6 h non-selective pre-enrichment). This level of sensitivity was attained even in the presence of high levels of other bacterial species that would otherwise have interfered with the results. With combinations of different antibodies, each with a unique fluorescent label, simultaneous analysis for two species was possible.

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Section: Introductionmentioning

confidence: 99%

Detection of low levels of specific Salmonella species by fluorescent antibodies and flow cytometry

McClelland

Pinder

1994

Journal of Applied Bacteriology

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“…Third, there is reduced scattering of 488 nm incident light for a nanorod of this size, minimizing changes in forward and side scattering at this wavelength, such that this labeling will not significantly interfere with complementary measurements of cell size and granularity by flow cytometry. Fourth, the gold nanorods chosen for this study have very large scattering cross-sections at their plasmon resonance wavelength, making them ideal tags compared with other nanoparticles used in previous studies, which were not matched to the excitation wavelength (15,17). Finally, gold nanorod markers can be fabricated to scatter across a broad spectral range, up to 2,000 nm, unlike gold nanosphere markers, which are limited to the visible spectrum.…”

Section: Discussionmentioning

confidence: 99%

“…Plasmonic coupling between nanosphere clusters most likely accounted for the small, detected enhancement, but required a significant increase in nanosphere concentration to observe the increased signal. Smaller immunolabeled gold nanospheres of 1 and 30 nm diameters were also investigated as scattering labels in flow cytometry using a 488-nm excitation source (17). For similar reasons, these experiments with labeled lymphocytes did not show a difference in light scattering between receptor-positive cells and controls.…”

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confidence: 99%

Plasmonic flow cytometry by immunolabeled nanorods

Crow¹,

Marinakos²,

Cook³

et al. 2010

Cytometry Pt A

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Fluorescence-based flow cytometry measures multiple cellular characteristics, including levels of receptor expression, by assessing the fluorescence intensity from a population of cells whose cell surface receptors are bound by a fluorescently labeled antibody or ligand for that receptor. Functionalized noble metal nanoparticles provide a complementary method of receptor labeling based on plasmonics for population analysis by flow cytometry. The potential benefits of using plasmonic nanoparticles to label cell surface receptors in flow cytometry include scattering intensity from a single particle that is equivalent to fluorescence intensity of 10 5 fluorescein molecules, biocompatibility and low cytotoxicity, and nonquenching optical properties. The large spectral tunability of nanorods also provides convenient access to plasmonic markers with peak surface plasmon resonances ranging from 600 to 2,200 nm, unlike gold nanosphere markers that are limited to visible wavelengths. Gold nanorod-based plasmonic flow cytometry is demonstrated herein by comparing the scattering of cells bound to antiepidermal growth factor receptor (EGFR)-conjugated nanorods to the emission of cells bound to anti-EGFR-conjugated fluorescent labels. EGFR-expressing cells exhibited a statistically significant six-fold increase in scattering when labeled with anti-EGFRconjugated nanorods compared with labeling with IgG1-conjugated nanorods. Large scattering intensities were observed despite using a 1,000-fold lower concentration of nanorod-conjugated antibody relative to the fluorescently labeled antibody. ' 2010International Society for Advancement of Cytometry Key terms flow cytometry; nanorod; plasmonic markers; epidermal growth factor receptor NOBLE metal nanoparticles exhibit unique optical properties that can be exploited for a variety of applications. These properties arise from the interaction of an incident electromagnetic (EM) field with free conduction band electrons in the nanoparticle. The conduction electrons are polarized relative to the heavier positive core ions because of the exciting EM field (1). The charge separation results in a restoring force and subsequent coherent oscillation of the conduction band electrons, sharply enhancing scattering and absorption at specific resonance frequencies. The scattering spectra of nanoparticles are dictated by intrinsic parameters such as their size, shape, and composition, as well as extrinsic factors such as the local refractive index of the surrounding medium. These intrinsic parameters enable the design of gold nanoparticles with peak resonances that can be tuned to specific wavelengths across a relatively wide spectral region (600-2,200 nm) simply by modifying the synthesis protocol to realize the appropriate structural parameters (2,3). Because slight changes in the local refractive index of nanoparticles result in a peak wavelength shift, noble metal nanoparticles are also exquisitely sensitive reporters of their local, nanoscale environment.The unique properties of p...

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“…Biomedical applications of nanoparticles (NP) involve their interaction with living cells through various mechanisms [1][2][3][4][5][6][7][8][9][10][11][12][13][14]. The most common tasks related to imaging nanoparticles during their interaction with living cells include such as to image and detect particles, to determine their location in individual cells and degree of aggregation Quantitative analysis (including imaging and monitoring) of NP structures in living cells is the challenge for the most of available imaging methods.…”

Section: Introductionmentioning

confidence: 99%

Methods for monitoring and imaging nanoparticles in cells

Lapotko

Lukianova

Чижик

2007

Nanoscale Imaging, Spectroscopy, Sensing, and Actuation for Biomedical Applications IV

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Successful targeting nanoparticles (NP) to specific cells requires reliable feedback about NP accumulation in cells. This task is a challenge for all optical methods due the size of NP and diffraction limit of optical devices. We modified several microscopy-based techniques for imaging and measuring NP in individual cells: photothermal, fluorescent, electron and atomic-force microscopies and flow cytometry. All those techniques were applied for quantitative analysis and imaging of interaction of gold NP (10 and 30 nm) with living tumor cells. Based on experimental results we performed comparison of all methods in terms of sensitivity, speed, sample requirements etc. We have found that standard microscopes may detect NP and their clusters in individual living cells through imaging NP-related thermal, fluorescent and other phenomena.

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Detection of triple antibody-binding lymphocytes in standard single laser flow cytometry using colloidal gold, fluorescein and phycoerythrin as labels

Cited by 28 publications

References 12 publications

Detection of low levels of specific Salmonella species by fluorescent antibodies and flow cytometry

Detection of low levels of specific Salmonella species by fluorescent antibodies and flow cytometry

Plasmonic flow cytometry by immunolabeled nanorods

Methods for monitoring and imaging nanoparticles in cells

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