Slide‐based cytometry for cytomics—A minireview

Tárnok, Attila

doi:10.1002/cyto.a.20317

Cited by 44 publications

(40 citation statements)

References 74 publications

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“…However, this approach is conventionally limited to adherent cells that are followed in time on the microscope stage (7) or cells in semisolid media. Cells that exist in suspension such as blood cells, monoclonal antibody producing CHO cells, or yeast cells must be attached to a slide for analysis.…”

mentioning

confidence: 99%

Single‐cell tracking with a reversing flow cytometer

Sitton

Srienc

2010

Cytometry Pt A

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We have developed an instrument based on a flow cytometer platform that is capable of tracking individual, suspended cells over extended time periods. The instrument repeatedly moves in a capillary the same volume segment of fluid containing tens to hundreds of suspended cells through the focal point of a laser. Individual cells are then tracked based on the timing of when they cross the laser, and cell properties are measured as in a conventional flow cytometer. Because cells are repeatedly measured the single-cell rates of change can be determined. The developed instrumentation was applied to measure the variability of ABC transporter activity in a population of human cancer cells and the temperature dependence of constitutively expressed Gfp in yeast. A wide range of transport rates can be observed in the cancer cell population while the single-cell Gfp fluorescence in yeast shows pronounced oscillations in response to temperature shifts. These observations are not detectable at the population level. Therefore, such measurements are useful for investigating cell function as they reveal how variable properties of single cells change over time. ' 2010 International Society for Advancement of Cytometry Key termssingle cell dynamics; rates of change; drug resistance IN both medicine and biotechnology, understanding how variability of properties arises and changes over time is of fundamental importance. Variability of properties has been indicated in the progression of certain diseases (1) and in explaining the productivity of chemicals or pharmaceuticals with engineered cultures (2,3). This variability has been generally related to genetic differences, to different positions in the cell cycle, to the exposure of a heterogeneous environment, or to stochastic variations due to the low number of molecules in individual cells. While the variability in the single-cell composition can be readily evaluated using conventional flow cytometry, the variability in dynamic properties of cells that reflect the rate at which cells change, are rather difficult to obtain and few examples exist in the literature.To experimentally measure the rate of how properties change, one must measure a property using either an Eulerian or Lagrangian reference frame (4,5). In the Eulerian reference frame one measures the distribution of a property in a cell population at discrete time points. In contrast, in the Lagrangian reference frame one follows individual cells over time and observes how their properties change. The properties of the entire cell population are then obtained as the sum of contributions of the individually tracked cells. The two reference frames are analogous to the fluid mechanic problem of converting Eulerian observations to Lagrangian (4).In the Eulerian reference frame, flow cytometry is primarily used to generate experimental data. This instrument yields a snapshot of the cellular property distribution at different time points and cells are discarded after the measurement. The ability to accurately understand...

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confidence: 99%

Single‐cell tracking with a reversing flow cytometer

Sitton

Srienc

2010

Cytometry Pt A

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“…This clinical cell analyses led to the concept of predictive medicine by cytomics which became one important key application for FCM and SBC facilitating individualized therapy (8,9,17).…”

Section: New Developments and Applications In Flow Andmentioning

confidence: 99%

“…They cover the field of cytometric analyses of cells in tissues, culture or suspensions (9), and also allow for simultaneous collection of a multitude of data. In most of the cytometric measurements, the direct link of parameter data to individual cells is missing.…”

Section: Technology Developments Concerning Slidementioning

confidence: 99%

Cytomics and nanobioengineering

Pierzchalski¹,

Robitzki²,

Mittag³

et al. 2008

Cytometry Part B Clinical

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The finding that an individual's genome differs as much as by many million variants from that of the human reference assembly diminished the great enthusiasm that every disease could be predicted based on nucleotide polymorphisms. Even individual cells of an organ may be specifically equipped to perform specific tasks and that the information of individual cells in a cell system is key information to understand function or dysfunction. Therefore, cytomics received great attention during the last years as it allows to quantitatively and qualitatively analyzing great number of individual cells, cell constituents, and of their intracellular and functional interactions in a cellular system and also giving the concept of analysis of these data.Exhaustive data extraction from multiparametric assays and multiple tests are the prerequisite for prediction of drug toxicity. Cytomics, as novel approach for unsupervised data analysis give a chance to find the most predictive parameters, which describe best the toxicity of a chemical. Cytomics is intrinsically connected to drug development and drug discovery.Focused on small structures, nanobioengineering is the ideal partner of cytomics, the systems biological discipline for cell population analysis. Realizing the idea ''from the molecule to the patient'' develops and offers chemical compounds, proteins, and other biomolecules, cells as well as tissues as instruments and products for a wide variety of biotechnological and biomedical applications.The integrative nanobioengineering combining different disciplines of nanotechnology will promote the development of innovative therapies and diagnostic methods. It can improve the precision of the measurements with focus on single cell analysis. By nanobioengineering and whole body imaging techniques, cytomics covers the field from molecules through bacterial cells, eukaryotic tissues, and organs to small animal live analysis. Toxicological testing and medical drug development are currently strongly broadening. It harbors the promise to substantially impact on various fields of biomedicine, drug discovery, and predictive medicine.As the number of scientific data is rising exponentially, new data analysis tools and strategies like cytomics and nanobioengineering take a lead and get closer to application. Bionanoengineering may strongly support the quantitative data supply, thus strengthening the rationale for cytomics approach. q

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“…Since that time different systems were developed to push the boundaries of image cytometry. The slide-based systems include laser scanning cytometry (3,(12)(13)(14)(15), standard (16)(17)(18)(19), enhanced wide-field (6,(20)(21)(22)(23) and confocal fluorescent microscopes (4,5,24). FCM can also be extended with imaging capabilities (25)(26)(27).…”

Section: Introductionmentioning

confidence: 99%

Automated multichannel fluorescent whole slide imaging and its application for cytometry

Varga¹,

Ficsor²,

Kamaras³

et al. 2009

Cytometry Pt A

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Slide-based image cytometry (SBC) has several advantages over flow cytometry but it is not widely used because of its low throughput, complicated workflow, and high price. Fully automated microscopes became affordable with the advent of whole slide imaging (WSI) and they can be transformed into a cytometer. A MIRAX MIDI automated whole slide imager was used with metal-halide and light emitting diode (LED)-based fluorescent illumination, filter block changer, and a cooled monochrome charge coupled device camera. The MIRAX control software was further developed for fluorescent sample detection, autofocusing, multichannel digitization, and signal correction due to nonuniform illumination. Fluorescent calibration beads were used to verify the linearity of the system. The HistoQuant software package of the MIRAX viewer was used for image segmentation and quantitative analysis. The data was displayed by the histogram, scatter plot, and gallery functions of the same program. Fluorescent samples can be reliably detected, focused, and scanned. The measured integrated fluorescence showed linearity with exposure time and staining intensity. Automated fluorescent WSI with stable LED illumination and high-quality homogeneous fluorescent slides can be used conveniently for SBC. ' 2009 International Society for Advancement of Cytometry

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Slide‐based cytometry for cytomics—A minireview

Cited by 44 publications

References 74 publications

Single‐cell tracking with a reversing flow cytometer

Single‐cell tracking with a reversing flow cytometer

Cytomics and nanobioengineering

Automated multichannel fluorescent whole slide imaging and its application for cytometry

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