A morphological and immunolabeling study of freeze-substituted Bacteroides forsythus.
We used a rapid freezing and freeze-substitution technique without glutaraldehyde and OsO4 fixation for the electron microscopic immunocytochemical demonstration of the surface structure of Bacteroides forsythus, an anaerobic Gram-negative periodontopathogen. Cells were applied to pieces of filter paper and freeze-substituted by plunge-freezing in liquid propane, substituted in methanol containing 0.5% uranyl acetate, and infiltrated with LR White resin. The membrane ultrastructure of B. forsythus was preserved well, and the labeling density of the freeze-submitted cells was compared to a conventional processing method. Our results show the usefulness of the freeze-substitution method for immunohistochemical studies of B. forsythus.
Moriguchi
K
K
Department of Anatomy, School of Dentistry, Aichi-Gakuin University, Nagoya, Aichi 464-8650, Japan. keiichi@dpc.aichi-gakuin.ac.jp
Higuchi
N
N
Murakami
Y
Y
Yoshimura
F
F
Nakamura
H
H
Ohno
N
N
D001439:Bacteroides / Q000648:ultrastructure*; D002462:Cell Membrane / Q000648:ultrastructure*; D048429:Cell Size; D017110:Freeze Substitution / Q000379:methods*; D007150:Immunohistochemistry / Q000379:methods*; D008854:Microscopy, Electron / Q000379:methods*; D009942:Organometallic Compounds; D013499:Surface Properties; D016610:Tissue Embedding / Q000379:methods*; D014021:Tissue Preservation / Q000379:methods*
D009942:Organometallic Compounds; C005460:uranyl acetate