Journal of Histochemistry and Cytochemistry Priciples for Free Access to Science
  Search:   
    >> Advanced Search

Guidelines | Subscriptions | About | exPRESS - Current - Archive | Business Information | Contact

This Article
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Altman, L. G.
Right arrow Articles by Papermaster, D. S.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Altman, L. G.
Right arrow Articles by Papermaster, D. S.
Social Bookmarking
Add to CiteULike   Add to Complore   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati   Add to Twitter  
What's this?

Rapid embedding of tissues in Lowicryl K4M for immunoelectron microscopy

LG Altman, BG Schneider and DS Papermaster

Lowicryl K4M (K4M) was recently introduced as an embedding medium for immunocytochemistry at the electron microscope level (BL Armbruster, E Carlemalm, R Chiovetti, RM Garavito, JA Hobot, E Kellenberger, W Villiger (1982):J Microsc 126:77 and E Carlemalm, M Garavito, W Villiger (1982):J Microsc 126:123). While earlier protocols of fixation and embedding required 4-6 days, the present method has reduced the processing time by accelerating both dehydration of tissues and polymerization of K4M so that tissues can be prepared for sectioning within 4 hr. The immunocytochemical labeling density was quantitated in order to determine relative antigen preservation in tissues embedded by the accelerated protocol as compared to slower K4M embedding techniques and to tissues embedded in glutaraldehyde-cross-linked bovine serum albumin (BSA). Thin sections of Bufo marinus kidney were labeled with rabbit antibody to Na+,K+ATPase alpha chain catalytic subunit isolated from B. marinus kidney microsomes (M Girardet, K Geering, JM Frantes, D Geser, BC Rossier, JP Kraehenbuhl, C Bron (1981):Biochemistry 20:6684). B. marinus retinas were labeled with rabbit anti-opsin. After fixation in paraformaldehyde(3%)-glutaraldehyde(3%), tissues were washed in buffer, dehydrated in 50, 75, and 90% dimethyl-formamide (DMF, 10 min each); K4M:DMF, 1:2 (15 min); K4M:DMF, 1:1, (20 min); K4M (25 min); K4M (30 min) at room temperature and transferred in fresh K4M to BEEM capsules for exposure to ultraviolet light (GE 15 watt, Black-lite, 10 cm, 45 min or less) at 4 degrees C. Thin sections were labeled successively with antibody, biotinylated sheep anti-rabbit F(ab')2 and avidin-ferritin. Ferritin labeling densities were determined by point counting. High labeling densities were observed with both antibodies, equaling or exceeding levels of labeling by slower protocols or embedment in BSA.

Volume 32, Issue 11, pp. 1217-1223, 11/01/1984
Copyright © 1984 by The Histochemical Society


Add to CiteULike CiteULike   Add to Complore Complore   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Reddit Reddit   Add to Technorati Technorati   Add to Twitter Twitter    What's this?


This article has been cited by other articles:


Home page
 Plant Physiol.Home page
K. H. Lackey, P. M. Pope, and M. D. Johnson
Expression of 1L-Myoinositol-1-Phosphate Synthase in Organelles
Plant Physiology, August 1, 2003; 132(4): 2240 - 2247.
[Abstract] [Full Text] [PDF]

Home page
 J. Am. Soc. Nephrol.Home page
M. Rantanen, T. Palmen, A. Patari, H. Ahola, S. Lehtonen, E. Astrom, T. Floss, F. Vauti, W. Wurst, P. Ruiz, et al.
Nephrin TRAP Mice Lack Slit Diaphragms and Show Fibrotic Glomeruli and Cystic Tubular Lesions
J. Am. Soc. Nephrol., June 1, 2002; 13(6): 1586 - 1594.
[Abstract] [Full Text] [PDF]

Home page
 IOVSHome page
D. S. Papermaster
The Birth and Death of Photoreceptors : The Friedenwald Lecture
Invest. Ophthalmol. Vis. Sci., May 1, 2002; 43(5): 1300 - 1309.
[Full Text] [PDF]

Home page
 J. Histochem. Cytochem.Home page
S. Miscia, A. Di Baldassarre, A. Cataldi, R. A. Rana, R. Di Pietro, D. Bosco, A. Grilli, G. Amerio, and G. Sabatino
Immunocytochemical Localization of Phospholipase C Isozymes in Cord Blood and Adult T-lymphocytes
J. Histochem. Cytochem., July 1, 1999; 47(7): 929 - 936.
[Abstract] [Full Text]

Home page
 J Am Coll CardiolHome page
U. N. Riede, U. Forstermann, H. Drexler, and with the technical assistance of Brigitte Freudenb
Inducible nitric oxide synthase in skeletal muscle of patients with chronic heart failure
J. Am. Coll. Cardiol., October 1, 1998; 32(4): 964 - 969.
[Abstract] [Full Text] [PDF]

Home page
 J. Cell Sci.Home page
X Chen, E. Reynolds, G Ranganayakulu, and J. O'Donnell
A maternal product of the Punch locus of Drosophila melanogaster is required for precellular blastoderm nuclear divisions
J. Cell Sci., January 12, 1994; 107(12): 3501 - 3513.
[Abstract] [PDF]

Home page
 Arch OphthalmolHome page
J. C. Morrison, N. L. L'Hernault, J. A. Jerdan, and H. A. Quigley
Ultrastructural Location of Extracellular Matrix Components in the Optic Nerve Head
Arch Ophthalmol, January 1, 1989; 107(1): 123 - 129.
[Abstract] [PDF]



Guidelines | Subscriptions | About | exPRESS - Current - Archive | Business Information | Contact
The Journal of Histochemistry & Cytochemistry is owned, published, and licensed by The Histochemical Society © 1984

 
Purchase HCS Short Course Manual on HCS site