Microcolumn Separations and the Analysis of Single Cells

Kennedy, Robert T.; Oates, Mary D.; Cooper, Bruce R.; Nickerson, Beverly; Jorgenson, James W.

doi:10.1126/science.2675314

Cited by 299 publications

(195 citation statements)

References 50 publications

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“…These can include single-cell lysis and separation of analytes by capillary electrophoresis (CE) (see also "Sample preparation: a key challenge"). In an early study by Kennedy et al [36], amino acids could readily be analyzed in extracts from individual neuron cells obtained from snails. Naphthalene-2,3-dicarboxyaldehyde was used to label amino acids in the cell extract to enable analysis by CE with laser-induced-fluorescence (LIF) detection.…”

Section: Fluorescence Detectionmentioning

confidence: 99%

Analytical techniques for single-cell metabolomics: state of the art and trends

2010

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Single-cell metabolomics is an emerging field that addresses fundamental biological questions and allows one to observe metabolic phenomena in heterogeneous populations of single cells. In this review, we assess the suitability of different detection techniques and present considerations on sample preparation for single-cell metabolomics. Although targeted analysis of single cells can readily be conducted using fluorescent probes and optical instruments (microscopes, fluorescence detectors), a comprehensive metabolomic approach requires a powerful label-free method, such as mass spectrometry (MS). Mass-spectrometric techniques applied to study small molecules in single cells include electrospray MS, matrix-assisted laser desorption/ionization MS, and secondary ion MS. Sample preparation is an important aspect to be taken into account during further development of methods for single-cell metabolomics.

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Section: Fluorescence Detectionmentioning

confidence: 99%

Analytical techniques for single-cell metabolomics: state of the art and trends

2010

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“…This volume compares favorably with the nanoliter volumes reported for derivatization reactions in capillary-based chemical cytometry methods (1)(2)(3)(4)(5), although smaller volumes have been reported (26). The capacity to confine a lysing, reacting plug of cell and reagents is an advantage that motivated prior work by us (8) and others (18) to use valves formed by multilayer soft lithography to confine approximately nanoliter volumes.…”

Section: Integrated Microfluidic Chip For Manipulation and Analysis Omentioning

confidence: 68%

Chemical cytometry on a picoliter-scale integrated microfluidic chip

Wheeler

Zare

2004

Proc. Natl. Acad. Sci. U.S.A.

239

186

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“…Cells are particularly sensitive to their environments, and an accurate assay will depend upon the success of the culture technique. Before the design process even begins, three issues require a good deal of forethought: providing proper growth conditions (i.e., adding fresh media; removing wastes; preventing air bubbles; maintaining proper temperature, pH, CO 2 , and humidity levels; etc. ); harvesting cells; and transporting them from the culture chamber to the different microchip architectures.…”

Section: On-chip Cell Culturementioning

confidence: 99%

Chemical Analysis of Single Mammalian Cells with Microfluidics

Price¹,

Culbertson²

2007

Anal. Chem.

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of Single Mammalian Cells with Microfluidics C ells are the fundamental building blocks of life. All basic physiological functions of multicellular organisms reside ultimately in the cell. The misregulation of cellular physiology results in disease at the organism level. Thus, comprehending cell physiology is key to understanding and curing diseases. Many physiological processes can be studied using populations of cells. Others occur either on short timescales (e.g., kinase signaling cascades) or nonsynchronously (e.g., response to an external chemical gradient), so that taking a population average will not lead to an understanding of how the cellular chemistry occurs. These types of processes require single-cell analysis, and thus discretion must be exercised when deciding under which circumstances bulk versus singlecell analyses are more appropriate (1). In addition, many diseases like cancer start with a single cell; therefore, if one would like to find the rare mutations in populations of cells that herald the inception of a disease, then cells must be examined individually.Probing behavior at the single-cell level, however, is a very challenging task, primarily because of the small sample volume, the low abundance of material, and the fragile nature of the cell itself. Analyzing the contents of a single cell requires sensitive detection techniques and handling procedures that do not stress or damage it. Additionally, no proper blank exists that can be used, so truly quantitative studies are difficult. Intense interest in single-cell physiology, however, is driving the analytical and biomedical engineering fields to improve the technology for examining cells. One of the most popular and promising areas is lab-on-a-chip devices to manipulate and analyze single cells.Since Jorgenson's groundbreaking work in 1989, CE has been used to examine the contents of single cells (2-4). However, many procedures for cell injection and lysis are time-consuming, and accuracy and reproducibility rely heavily on the skill of the researcher. Furthermore, the limited number of architectures provided by microbore tubing and its relatively large volume restrict the types of processes that can be investigated. Conversely, microfluidics, or lab-on-a-chip technology, offers a versatile format in which biological cells can be analyzed.These miniaturized devices provide several analytical and operational advantages over conventional macroscale systems (5, 6). Microfluidic architectures provide precise spatial control over reagents and samples, are capable of fast analysis times, can be automated, and can precisely manipulate picoliter volumes of material without dilution. In addition, microfluidic systems are amenable to many different detection schemes, can be manufactured from many different materials at relatively low cost, allow flexibility of design, and provide the capability of integrating a series of multiple tasks (sample preparation, mixing, separation, etc.) in both serial and parallel schemes. Portable versions of these sys...

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Microcolumn Separations and the Analysis of Single Cells

Cited by 299 publications

References 50 publications

Analytical techniques for single-cell metabolomics: state of the art and trends

Analytical techniques for single-cell metabolomics: state of the art and trends

Chemical cytometry on a picoliter-scale integrated microfluidic chip

Chemical Analysis of Single Mammalian Cells with Microfluidics

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