High-resolution immunogold labeling of cell surfaces: The hypothetical homodimeric nature of the cd5 receptor

1988 ◽  
Vol 46 ◽  
pp. 380-381
Author(s):  
E. de Harven ◽  
H. Christensen
Keyword(s):  
Field Emission ◽  
Molecular Level ◽  
Human Peripheral Blood ◽  
Lanthanum Hexaboride ◽  
Peripheral Blood Lymphocytes ◽  
Cell Surfaces ◽  
Gold Particles ◽  
Human Peripheral Blood Lymphocytes ◽  
Backscattered Electron ◽  
Electron Imaging

Molecules exposed on cell surfaces and labeled with colloidal gold markers can be optimally demonstrated using the backscattered electron imaging (BEI) mode of the scanning electron microscope (SEM). Steric hindrance, however, limits labeling efficiency, making it necessary to use gold markers of small size for labeling at the molecular level. Using a JEOL 840 SEM equipped with a lanthanum hexaboride (LaB6) cathode, 13 nm gold particles were demonstrated. This, however, seems to represent the limit of the resolution of this type of instrument in the BEI mode. Fortunately, it has been demonstrated by Walther and Muller that 5 nm gold particles can be seen in the BEI mode, using field emission SEM.We have confirmed this observation, using the JEOL 890 field emission SEM and a solid state backscattered electron detector. Human peripheral blood lymphocytes prefixed with 0.1% glutaraldehyde, incubated with the murine monoclonal antibody LEU-1 (CD5), were labeled with a goat anti-murine IgG adsorbed on 5 nm gold particles (GAM-G5, Janssen Pharmaceutica, Beerse, Belgium) according to previously described procedures.

Quantification of cell-surface expressed antigenic sites with 5-nm gold markers: Getting close to biologically relevant data

1989 ◽  
Vol 47 ◽  
pp. 1050-1051
Author(s):  
Etienne de Harven ◽  
Hilary Christensen ◽  
Richard Leung ◽  
Cameron Ackerley
Keyword(s):  
Cell Surface ◽  
Human Peripheral Blood ◽  
Contour Length ◽  
Thin Sections ◽  
Gold Particles ◽  
Biologically Relevant ◽  
Transmission Electron ◽  
Entire Cell ◽  
Electron Imaging ◽  
Cell Contour

The T-derived subset of human peripheral blood normal lymphocytes has been selected as a model system to study the usefulness of 5 nm gold markers for quantification of single epitopes expressed on cell surfaces. The chosen epitopes are parts of the CD3 and CD5 molecules and can be specifically identified by hybridoma produced monoclonal antibodies (MoAbs; LEU-4 and LEU-1; Becton-Dick- inson, Mountain view, CA) . An indirect immunolabeling procedure, with goat anti-murine IgG adsorbed on the surface of 5 nm colloidal gold particles (GAM-G5, Janssen Pharmaceutica, Beerse, Belgium) has been used. Backscattered Electron Imaging (BEI) in a field emission scanning electronmicroscope (SEM) and transmission electron microscopy of thin sections of lymphocytes labeled before plastic embedding, were both used to identify and quantitate gold labeled cell surface sites, Estimating that the thickness of “silver” sections is approximately 60 nm and counting the number of gold particles on the entire cell perimeter, we calculated that, for LEU-4, the number of markers per um2 of cell surface is in the 140-160 range (Fig.l). Cell contour length measurements indicated that the surface of one lymphocyte is approximately 130-160 um2 that of a smooth sphere of identical diameter, reflecting the role of microvilli in expanding the surface area. The total number of gold labeled sites on the surface of one lymphocyte averages, therefore between 20,000 and 24,000 per cell.

scholarly journals Expression of Opioid Receptors in Cells of the Immune System

2020 ◽  
Vol 22 (1) ◽  
pp. 315
Author(s):  
Jana Brejchova ◽  
Vladimir Holan ◽  
Petr Svoboda
Keyword(s):  
Immune System ◽  
Opioid Receptors ◽  
Methadone Maintenance ◽  
Human Peripheral Blood ◽  
Peripheral Blood Lymphocytes ◽  
Immunomodulatory Effects ◽  
Human Peripheral Blood Lymphocytes ◽  
Opioid Drugs ◽  
Human Immune ◽  
In Cells

The observation of the immunomodulatory effects of opioid drugs opened the discussion about possible mechanisms of action and led researchers to consider the presence of opioid receptors (OR) in cells of the immune system. To date, numerous studies analyzing the expression of OR subtypes in animal and human immune cells have been performed. Some of them confirmed the expression of OR at both the mRNA and protein level, while others did not detect the receptor mRNA either. Although this topic remains controversial, further studies are constantly being published. The most recent articles suggested that the expression level of OR in human peripheral blood lymphocytes could help to evaluate the success of methadone maintenance therapy in former opioid addicts, or could serve as a biomarker for chronic pain diagnosis. However, the applicability of these findings to clinical practice needs to be verified by further investigations.

scholarly journals Analysis of P-glycoprotein-mediated membrane transport in human peripheral blood lymphocytes using the UIC2 shift assay

Cytometry ◽  
2003 ◽  
Vol 53A (2) ◽  
pp. 67-78 ◽  
Author(s):  
Suk W. Park ◽  
Noureddine Lomri ◽  
Luiz A. Simeoni ◽  
John P. Fruehauf ◽  
Eugene Mechetner
Keyword(s):  
Membrane Transport ◽  
Peripheral Blood ◽  
Human Peripheral Blood ◽  
Peripheral Blood Lymphocytes ◽  
Blood Lymphocytes ◽  
Human Peripheral Blood Lymphocytes ◽  
P Glycoprotein
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