Application of immunocryoultramicrotomy to free cells: ultrastructural localization of myeloperoxidase in neutrophils of human peripheral blood.
Immunocryoultramicrotomy is generally considered the most suitable method for observing the immunostainings by monoclonal antibodies to intracellular antigens by electron microscopy. However, this procedure with the free cells such as peripheral blood cells or cultured cells is not widely used because of the difficulty in preparing ultrathin frozen sections. We report here a new technique for making ultrathin frozen sections of peripheral blood cells by positioning the cell in a semigelatinized medium for ultrasectioning. Monoclonal antibody against myeloperoxidase (MPO) was reacted with the ultrathin sections made with this new technique to observe the ultrastructural localization of intracellular MPO. MPO was observed in the primary granules of the neutrophils.
Saito
N
N
Department of Internal Medicine, Asahikawa Medical College, Hokkaido, Japan.
Takemori
N
N
Hirai
K
K
Kubota
N
N
Onodera
R
R
Namiki
M
M
D000911:Antibodies, Monoclonal; D001773:Blood Cells / Q000201:enzymology* / Q000648:ultrastructure; D016616:Cryoultramicrotomy; D006801:Humans; D016253:Microscopy, Immunoelectron / Q000379:methods*; D008867:Microtomy / Q000379:methods*; D009504:Neutrophils / Q000201:enzymology* / Q000648:ultrastructure; D009195:Peroxidase / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D009195:Peroxidase