skip to main content
OSTI.GOV title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Chemical analysis of individual mammalian cells

Abstract

The extremely small size of mammalian cells creates an unusual challenge for the analytical chemist, both in terms of separation and detection. Under a microscope, it is possible to confirm the injection of individual cells such as erythrocyte into capillaries with 10-{mu}m i.d. by hydrostatic pressure. The ionic contents can then be separated by capillary electrophoresis after the cell lyses. Enzymes at the zeptomole level can be monitored by on-column fluorescence enzyme assay. On-column particle-counting immunoassay can be applied to a broad range of analytes (antigens), also at the zeptomole level. The authors report here the simultaneous determination of the amounts of glucose-6-phosphate dehydrogenase (G6PDH) and their activities in individual erythrocytes by using a combination of the two detection schemes. Insights into the degradation of proteins as a function of cell age can be derived.

Authors:
;  [1]
  1. Ames Lab., IA (United States)
Publication Date:
OSTI Identifier:
191665
Report Number(s):
CONF-941098-
TRN: 95:006733-0021
Resource Type:
Conference
Resource Relation:
Conference: FACSS XXI: 21st annual conference of the Federation of Analytical Chemistry and Spectroscopy Societies (FACSS), St. Louis, MO (United States), 2-7 Oct 1994; Other Information: PBD: 1994; Related Information: Is Part Of 21st annual conference of the Federation of Analytical Chemistry and Spectroscopy Societies; PB: 257 p.
Country of Publication:
United States
Language:
English
Subject:
55 BIOLOGY AND MEDICINE, BASIC STUDIES; 40 CHEMISTRY; ANIMAL CELLS; CHEMICAL ANALYSIS; ENZYMES; QUANTITATIVE CHEMICAL ANALYSIS; FLUORESCENCE SPECTROSCOPY; ERYTHROCYTES; ELECTROPHORESIS

Citation Formats

Tan, W., and Yeung, E.S.. Chemical analysis of individual mammalian cells. United States: N. p., 1994. Web.
Tan, W., & Yeung, E.S.. Chemical analysis of individual mammalian cells. United States.
Tan, W., and Yeung, E.S.. 1994. "Chemical analysis of individual mammalian cells". United States. doi:.
@article{osti_191665,
title = {Chemical analysis of individual mammalian cells},
author = {Tan, W. and Yeung, E.S.},
abstractNote = {The extremely small size of mammalian cells creates an unusual challenge for the analytical chemist, both in terms of separation and detection. Under a microscope, it is possible to confirm the injection of individual cells such as erythrocyte into capillaries with 10-{mu}m i.d. by hydrostatic pressure. The ionic contents can then be separated by capillary electrophoresis after the cell lyses. Enzymes at the zeptomole level can be monitored by on-column fluorescence enzyme assay. On-column particle-counting immunoassay can be applied to a broad range of analytes (antigens), also at the zeptomole level. The authors report here the simultaneous determination of the amounts of glucose-6-phosphate dehydrogenase (G6PDH) and their activities in individual erythrocytes by using a combination of the two detection schemes. Insights into the degradation of proteins as a function of cell age can be derived.},
doi = {},
journal = {},
number = ,
volume = ,
place = {United States},
year = 1994,
month =
}

Conference:
Other availability
Please see Document Availability for additional information on obtaining the full-text document. Library patrons may search WorldCat to identify libraries that hold this conference proceeding.

Save / Share:
  • This paper describes initial experiments to perform laser ablation mass spectrometry in real time on airborne microparticles. The microparticles are sampled directly from the air by a particle inlet system into the vacuum chamber of a mass spectrometer. An incoming particle is detected as it passes through two CW laser beams and a pulsed laser is triggered to intercept the particle for laser ablation/ionization in the mass spectrometer. The initial studies were made with an existing ion trap mass spectrometer with the particle sampling occurring at the center of the trap electrodes. Performance of the inlet system, particle detection, andmore » preliminary results are described.« less
  • An improved dual-laser flow cytometric system for quantitative analysis and sorting of mammalian cells has been developed using a low-power argon and high-power krypton laser as illumination sources, thus permitting the excitation of fluorescent dyes having absorption regions ranging from the ultraviolet to infrared. Cells stained in liquid suspension with fluorescent dyes enter a flow chamber where they intersect two spatially separated laser beams. Separate pairs of quartz beam-shaping optics focus each beam onto the cell stream. Electro-optical sensors measure fluorescence and light scatter from cells that are processed electronically and displayed as frequency distribution histograms. Cells also can bemore » electronically separated and microscopically identified. The ease and versatility of operation designed into this system represent a marked technological improvement for dual-laser excited flow systems. Details of this instrument are described along with illustrative examples of cells stained with mithramycin and rhodamine and analyzed for DNA content, total protein, and nuclear and cytoplasmic diameter.« less
  • The authors have developed a test procedure to detect, without electrical access to individual cells, defective cells in a module: the full array is illuminated by a permanent beam, which allows self-biassing of the cells. A modulated beam illuminates the cell under test, thus generating an ac signal, function of the dynamical impedance, and hence of the cell bias. This double illumination technique enhances the discrimination ratio between normal and defective cells: in a series array of 25 cells, a cell with a I /SUB sc/ lower by 10%, yields, at the module terminals, a signal 200 times larger thanmore » a normal cell. Similar results have been obtained with other configurations (series-parallel) and confirmed by numerical modelling.« less
  • We have developed a cell-mediated mutagenesis assay in which cells with the appropriate markers for mutagenesis are co-cultivated with either lethally irradiated rodent embryonic cells that can metabolize carcinogenic hydrocarbons or with primary rat liver cells that can metabolize chemicals carcinogenic to the liver. During co-cultivation, the reactive metabolites of the procarcinogen appear to be transmitted to the mutable cells and induce mutations in them. Assays of this type make it possible to demonstrate a relationship between carcinogenic potency of the chemicals and their ability to induce mutations in mammalian cells. In addition, by simultaneously comparing the frequencies of transformationmore » and mutation induced in normal diploid hamster cells by benzo(a)pyrene (BP) and one of its metabolites, it is possible to estimate the genetic target size for cell transformation in vitro.« less
  • It has been shown that most carcinogens require metabolic activation to electrophilic reactants prior to exerting their toxic effects. Such reactive metabolites are capable of binding to cellular macromolecules including DNA, suggesting the production of critical mutations as the initial event in the process of tumor development. Indeed most chemical carcinogens have been shown to act as mutagens as well. For an accurate assessment of the genetic risk from exposure to these chemicals, it is not sufficient to merely indicate mutagenic potential. It must be shown that a quantitative relationship exists between carcinogenicity and mutagenicity and that the same metabolitesmore » are responsible in both cases. In order to make a valid extrapolation from the in vitro systems to man, one must devise systems which should also take into account the organ and/or species specificity of the carcinogens. Some classes of chemicals do not appear to act via a mutational mechanism and yet can influence the development of tumors. These chemicals are thought to act subsequent to an initiation event caused by another chemical and amplify or promote the development of tumors. It would be useful to devise simple in vitro systems in which both the identity and mode of action of such chemicals can be investigated. The results obtained from such an approach and the conclusions drawn from these studies are discussed. It was found that the human cell lines, HL-60 and HO melanoma, may offer a useful model for studying the mode of action of certain classes of tumor promoters and in evaluating some unknown chemicals for this activity. The cell-mediated mutagenesis assays enable the mutagenic and hence possibly, the initiating activity of a chemical to be investigated, whereas specific alterations in the growth and cell differentiation of human cells such as the HL-60 myeloid cells produced by a chemical may be indicative of a tumor-promoting potential. (ERB)« less