Imaging Flow Cytometry

Barteneva, Natasha S.; Fasler‐Kan, Elizaveta; Vorobjev, Ivan A.

doi:10.1369/0022155412453052

Cited by 168 publications

(87 citation statements)

References 93 publications

(150 reference statements)

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“…At the same time, because of flow cytometry’s successful applications in immunology and thanks to the growing appreciation of researchers for the complexity of the immune system, further technological advances has focused on satisfying the increasing need for polychromatic approaches to flow cytometry; researchers have developed flow cytometry to simultaneously measure 19 parameters—17 fluorescence and 2 scatter parameters in a high-speed manner 7 . The broadly useful technology—flow cytometry—has been evolving slowly until imaging flow cytometry (IFC) became a resurgence of interest in the past decade 8,9 . Due to its high-throughput and multiparametric analysis, by supporting detection of single cell properties at rates from hundreds to 100,000 cells per second, conventional flow cytometry is an irreplaceable cytologic instrumentation when study of high-volume cell populations and subpopulations needs to be performed 10–12 .…”

Section: Introductionmentioning

confidence: 99%

Review: imaging technologies for flow cytometry

et al. 2016

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High throughput single cell imaging is a critical enabling and driving technology in molecular and cellular biology, biotechnology, medicine and related areas. Imaging flow cytometry combines single-cell imaging capabilities of microscopy with the high-throughput capabilities of conventional flow cytometry. Recent advances in imaging flow cytometry are remarkably revolutionizing the single-cell analysis. This article describes recent imaging flow cytometry technologies and their challenges.

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Section: Introductionmentioning

confidence: 99%

Review: imaging technologies for flow cytometry

et al. 2016

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“…It combines the features of classic flow cytometry and fluorescent microscopy with advanced data-processing algorithms that allows researchers to analyze individual cells for protein expression/localization/translocation as well as morphological features of single cells-correlating various endpoints within a homogenous population, moving beyond simple population averages. Moreover, since each acquired cell is visualized, non-specifically stained false positives can be avoided as well as non-viable cells dismissed (Barteneva et al 2012).…”

Section: Introductionmentioning

confidence: 99%

Myogenic progenitors and imaging single-cell flow analysis: a model to study commitment of adult muscle stem cells

Trapečar

Kelc

Gradišnik

et al. 2014

J Muscle Res Cell Motil

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Research on skeletal muscles suffers from a lack of appropriate human models to study muscle formation and regeneration on the regulatory level of single cells. This hampers both basic understanding and the development of new therapeutic approaches. The use of imaging multicolour flow cytometry and myogenic stem cells can help fill this void by allowing researchers to visualize and quantify the reaction of individual cultured cells to bioactives or other physiological impulses. As proof of concept, we subjected human CD56+ satellite cells to reference bioactives follistatin and Malva sylvestris extracts and then used imaging multicolor flow cytometry to visualize the stepwise activation of myogenic factors MyoD and myogenin in individual cells. This approach enabled us to evaluate the potency of these bioactives to stimulate muscle commitment. To validate this method, we used multi-photon confocal microscopy to confirm the potential of bioactives to stimulate muscle differentiation and expression of desmin. Imaging multicolor flow cytometry revealed statistically significant differences between treated and untreated groups of myogenic progenitors and we propose the utilization of this concept as an integral part of future muscle research strategies.

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“…A recently introduced technique, imaging flow cytometry (IFC) delivers both FACS data and microscopical images of cells. 9,10 While it is very convenient to obtain visual data of the same cells, which are analyzed in FACS, it comes at the cost of lower sample throughput compared to conventional FACS and lower resolution compared to CLSM. IFC has been used to quantify internalization, but requires to permeabilize cells and co-stain internal cell compartments, thereby only measuring particles within the stained cell compartments.…”

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confidence: 99%

Precise Quantification of Nanoparticle Internalization

et al. 2013

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Nanoparticles have opened new exciting avenues for both diagnostic and therapeutic applications in human disease, and targeted nanoparticles are increasingly used as specific drug delivery vehicles. The precise quantification of nanoparticle internalization is of importance to measure the impact of physical and chemical properties on the uptake of nanoparticles into target cells or into cells responsible for rapid clearance. Internalization of nanoparticles has been measured by various techniques, but comparability of data between different labs is impeded by lack of a generally accepted standardized assay. Furthermore, the distinction between associated and internalized particles has been a challenge for many years, although this distinction is critical for most research questions. Previously used methods to verify intracellular location are typically not quantitative and do not lend themselves to high throughput analysis. Here we developed a mathematical model which integrates the data from high throughput flow cytometry measurements with data from quantitative confocal microscopy. The generic method described here will be a useful tool in biomedical nanotechnology studies. The method was then applied to measure the impact of surface coatings of vesosomes on their internalization by cells of the reticuloendothelial system (RES). RES cells are responsible for rapid clearance of nanoparticles, and the resulting fast blood clearance is one of the major challenges in biomedical applications of nanoparticles. Coating of vesosomes with long chain polyethylene glycol showed a trend for lower internalization by RES cells.

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Imaging Flow Cytometry

Cited by 168 publications

References 93 publications

Review: imaging technologies for flow cytometry

Review: imaging technologies for flow cytometry

Myogenic progenitors and imaging single-cell flow analysis: a model to study commitment of adult muscle stem cells

Precise Quantification of Nanoparticle Internalization

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