Detection of mammosomatotroph cells and identification of the coexistence of growth hormone and prolactin within the same secretory granules in these cells using confocal laser scanning microscopy.
This study was designed to examine whether mammosomatotroph cells (MS cells) can be easily detected using confocal laser scanning microscopy (CLSM) and whether the coexistence of growth hormone (GH) and prolactin (PRL) within the same secretory granule can be identified in the MS cell using CLSM. Conventional epoxy resin-embedded tissues of mixed GH- and PRL-secreting human pituitary adenomas were used for this double-labelling immunofluorescent study by CLSM. A semithin section of the tissue after plastic removal and bleaching was immunohistochemically double-stained with primary antibodies against GH and PRL, followed by secondary antibodies conjugated with Rhodamine (GH) and FITC (PRL). MS cells simultaneously showing fluorescence of both Rhodamine and FITC were easily detected by CLSM at lower magnification. At higher magnification, the coexistence of Rhodamine and FITC on the same secretory granule was identified by using a superimposed display. This finding was confirmed by immunoelectron microscopy. The CLSM technique may be useful for the study of MS cells.
Kabuto
M
M
Department of Neurosurgery, Fukui Medical School, Japan.
Kubota
T
T
Kobayashi
H
H
Nakagawa
T
T
Kitai
R
R
Kodera
T
T
D000236:Adenoma; D003594:Cytoplasmic Granules / Q000737:chemistry* / Q000648:ultrastructure; D005455:Fluorescent Antibody Technique; D013006:Growth Hormone / Q000032:analysis*; D006801:Humans; D018613:Microscopy, Confocal; D016253:Microscopy, Immunoelectron; D010902:Pituitary Gland / Q000473:pathology; D011388:Prolactin / Q000032:analysis*; D014407:Tumor Cells, Cultured / Q000166:cytology / Q000648:ultrastructure
D011388:Prolactin; D013006:Growth Hormone