|
|
|
|
 |
|||
|
|||
Microwave fixation improves antigenicity of glutaraldehyde-sensitive antigens while preserving ultrastructural detail
MC Jamur, CD Faraco, LO Lunardi, RP Siraganian and C Oliver
Departamento de Biologia Celular, Universidade Federal do Parana, Curitiba, Pr, Brasil.
Microwave fixation for electron microscopy has been used primarily for post-embedding immunocytochemistry. The present study examined the ability of microwave fixation to preserve the antigenicity of glutaraldehyde-sensitive antigens for pre-embedding immunocytochemistry. Five monoclonal antibodies (MAbs) directed against cell surface components of rat mast cells were tested. The MAbs failed to show any labeling of conventionally fixed rat bone marrow-derived mast cells even at glutaraldehyde concentrations as low as 0.1%. Strong staining of mast cell plasma membranes was seen when bone marrow was initially fixed with 2% formaldehyde and then refixed in 2% glutaraldehyde/2% formaldehyde after immunostaining. However, the ultrastructural preservation of the cells was poor. Antigenicity and morphological detail were both preserved when bone marrow was fixed in 0.05% glutaraldehyde/2% formaldehyde for 4 sec in a 550-W microwave oven. With this method, mast cells in various stages of maturation as well as cells that did not contain granules were immunoreactive. This method should prove useful with antigens from many different cell types that are sensitive to glutaraldehyde fixation.
Volume 43,
Issue 3,
pp. 307-311,
03/01/1995
Copyright © 1995 by The Histochemical Society
CiteULike 
Complore 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati What's this?
This article has been cited by other articles:
|
|
 |
|
  C. E. Gagna, H.-R. Kuo, N. J. Chan, E. J. Mitacek, A. Spivak, T. D. Pasquariello, C. Balgobin, R. Mukhi, and W. C. Lambert Novel DNA Staining Method and Processing Technique for the Quantification of Undamaged Double-stranded DNA in Epidermal Tissue Sections by PicoGreen Probe Staining and Microspectrophotometry J. Histochem. Cytochem., October 1, 2007; 55(10): 999 - 1014. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
V. E. Arana-Chavez and A. Nanci High-resolution Immunocytochemistry of Noncollagenous Matrix Proteins in Rat Mandibles Processed with Microwave Irradiation J. Histochem. Cytochem., September 1, 2001; 49(9): 1099 - 1110. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. C. Jamur, A. C. G. Grodzki, A. N. Moreno, L. de Fátima C. de Mello, M. V. D. Pastor, E. H. Berenstein, R. P. Siraganian, and C. Oliver Identification and Isolation of Rat Bone Marrow-derived Mast Cells Using the Mast Cell-specific Monoclonal Antibody AA4 J. Histochem. Cytochem., February 1, 2001; 49(2): 219 - 228. [Abstract] [Full Text]
|
|
|
|
|
|
M. C. Jamur, A. C. G. Grodzki, A. N. Moreno, W. D. Swaim, R. P. Siraganian, and C. Oliver Immunomagnetic Isolation of Rat Bone Marrow-derived and Peritoneal Mast Cells J. Histochem. Cytochem., December 1, 1997; 45(12): 1715 - 1722. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Grutzkau, S. Kruger-Krasagakes, H. Kogel, A. Moller, U. Lippert, and B. M. Henz Detection of Intracellular Interleukin-8 in Human Mast Cells: Flow Cytometry as a Guide for Immunoelectron Microscopy J. Histochem. Cytochem., July 1, 1997; 45(7): 935 - 946. [Abstract] [Full Text] [PDF]
|
|
| Guidelines | Subscriptions | About | exPRESS - Current - Archive | Business Information | Contact |