Parallel Imaging Microfluidic Cytometer

Ehrlich, D. J.; McKenna, Brian; Evans, James G.; Belkina, Anna C.; Denis, Gerald V.; Sherr, David H.; Cheung, Monit

doi:10.1016/b978-0-12-374912-3.00003-1

Cited by 6 publications

(2 citation statements)

References 30 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…It allows discovery of changes in cell physiology (e.g., in combination with microscopic screening [131,132]), and also screening for bacteria [133] or enzyme activities [134]. A limitation is the high cost of these advanced instruments and the fact that they require a highly trained operator and strict laboratory conditions.…”

Section: Fig 10 Lymphocyte Transformation Test (Ltt)mentioning

confidence: 99%

Immunodiagnostics and immunosensor design (IUPAC Technical Report)

Gubala

Klein²,

Templeton

et al. 2014

Pure and Applied Chemistry

View full text Add to dashboard Cite

This work compiles information on the principles of diagnostic immunochemical methods and the recent advances in this field. It presents an overview of modern techniques for the production of diagnostic antibodies, their modification with the aim of improving their diagnostic potency, the different types of immunochemical detection systems, and the increasing diagnostic applications for human health that include specific disease markers, individualized diagnosis of cancer subtypes, therapeutic and addictive drugs, food residues, and environmental contaminants. A special focus lies in novel developments of immunosensor techniques, promising approaches to miniaturized detection units and the associated microfluidic systems. The trends towards high-throughput systems, multiplexed analysis, and miniaturization of the diagnostic tools are discussed. It is also made evident that progress in the last few years has largely relied on novel chemical approaches.

show abstract

Section: Fig 10 Lymphocyte Transformation Test (Ltt)mentioning

confidence: 99%

Immunodiagnostics and immunosensor design (IUPAC Technical Report)

Gubala

Klein²,

Templeton

et al. 2014

Pure and Applied Chemistry

View full text Add to dashboard Cite

show abstract

“…Owing to their advantages in integration and multiplexing, microfluidic platforms have attracted significant attention in cellular analysis. 11 Microfluidic platforms have been developed for many unit functions such as cell culture, [12][13][14] flow cytometry, [15][16][17][18][19] cell sorting [20][21][22] and single cell imaging. [23][24][25][26] We recently reported on chip-based flow cytometry for measurement of genes (by fluorescence in situ hybridization) 27 and proteins (by immunostaining) in single cells.…”

Section: Introductionmentioning

confidence: 99%

Microfluidically-unified cell culture, sample preparation, imaging and flow cytometry for measurement of cell signaling pathways with single cell resolution

Perroud

Srivastava

et al. 2012

Lab Chip

View full text Add to dashboard Cite

We have developed a microfluidic platform that enables, in one experiment, monitoring of signaling events spanning multiple time-scales and cellular locations through seamless integration of cell culture, stimulation and preparation with downstream analysis. A combination of two single-cell resolution techniques-on-chip multi-color flow cytometry and fluorescence imaging provides multiplexed and orthogonal data on cellular events. Automated, microfluidic operation allows quantitatively- and temporally-precise dosing leading to fine time-resolution and improved reproducibility of measurements. The platform was used to profile the toll-like receptor (TLR4) pathway in macrophages challenged with lipopolysaccharide (LPS)-beginning with TLR4 receptor activation by LPS, through intracellular MAPK signaling, RelA/p65 translocation in real time, to TNF-α cytokine production, all in one small macrophage population (< 5000 cells) while using minute reagent volume (540 nL/condition). The platform is easily adaptable to many cell types including primary cells and provides a generic platform for profiling signaling pathways.

show abstract

A computational platform for robotized fluorescence microscopy (I): High‐content image‐based cell‐cycle analysis

Furia¹,

Pelicci²,

Faretta³

2013

Cytometry Pt A

View full text Add to dashboard Cite

Hardware automation and software development have allowed a dramatic increase of throughput in both acquisition and analysis of images by associating an optimized statistical significance with fluorescence microscopy. Despite the numerous common points between fluorescence microscopy and flow cytometry (FCM), the enormous amount of applications developed for the latter have found relatively low space among the modern high-resolution imaging techniques. With the aim to fill this gap, we developed a novel computational platform named A.M.I.CO. (Automated Microscopy for Image-Cytometry) for the quantitative analysis of images from widefield and confocal robotized microscopes. Thanks to the setting up of both staining protocols and analysis procedures, we were able to recapitulate many FCM assays. In particular, we focused on the measurement of DNA content and the reconstruction of cell-cycle profiles with optimal parameters. Standard automated microscopes were employed at the highest optical resolution (200 nm), and white-light sources made it possible to perform an efficient multiparameter analysis. DNA-and protein-content measurements were complemented with image-derived information on their intracellular spatial distribution. Notably, the developed tools create a direct link between image-analysis and acquisition. It is therefore possible to isolate target populations according to a definite quantitative profile, and to relocate physically them for diffraction-limited data acquisition. Thanks to its flexibility and analysis-driven acquisition, A.M.I.CO. can integrate flow, image-stream and laser-scanning cytometry analysis, providing high-resolution intracellular analysis with a previously unreached statistical relevance. ' 2013 International Society for Advancement of Cytometry Key termsimage-cytometry; cell-cycle; automated microscopy; image analysis LIMITED sample availability, as for material of clinical origin, and a poor statistical representation of targeted cell populations, for example, stem cells, can make the comprehension of biological heterogeneity a very challenging task (1-4). High-content approaches have been consequently developed to extend the number of simultaneously analyzable parameters (5-7).Flow cytometry (FCM) is a powerful technique that provides statistically relevant measurements. Quantification at single-cell level, elevated content, fast acquisition, and contained analysis time make it a very valuable tool for the identification of rare cell populations. In addition, enormous advances in limiting the required amount of sample per analysis have also been achieved by system miniaturization (7-11).This excellent "statistical resolution" is, however, coupled to a complete absence of an intracellular view and, to date, fluorescence microscopy maintains its leadership in providing a high-resolution description of the inner cell compartments. Nevertheless, technological efforts have tried to fill the imaging gap between FCM and fluorescence microscopy, leading to the development of ...

show abstract

Parallel Imaging Microfluidic Cytometer

Cited by 6 publications

References 30 publications

Immunodiagnostics and immunosensor design (IUPAC Technical Report)

Immunodiagnostics and immunosensor design (IUPAC Technical Report)

Microfluidically-unified cell culture, sample preparation, imaging and flow cytometry for measurement of cell signaling pathways with single cell resolution

A computational platform for robotized fluorescence microscopy (I): High‐content image‐based cell‐cycle analysis

Contact Info

Product

Resources

About