Kabuto M, Kubota T, Kobayashi H, Nakagawa T, Kitai R, Kodera T
Department of Neurosurgery, Fukui Medical School, Japan.
Histochem Cell Biol. 1995 Nov;104(5):363-7. doi: 10.1007/BF01458130.
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.
本研究旨在探讨是否可通过共聚焦激光扫描显微镜(CLSM)轻松检测到乳腺生长激素细胞(MS细胞),以及是否能够利用CLSM在MS细胞中识别生长激素(GH)和催乳素(PRL)在同一分泌颗粒中的共存情况。采用常规环氧树脂包埋的分泌GH和PRL的人垂体混合腺瘤组织,进行此项CLSM双标记免疫荧光研究。去除塑料包埋剂并漂白后的组织半薄切片,用抗GH和PRL的一抗进行免疫组织化学双重染色,随后用与罗丹明(针对GH)和异硫氰酸荧光素(针对PRL)偶联的二抗进行染色。通过CLSM在较低放大倍数下可轻松检测到同时显示罗丹明和异硫氰酸荧光素荧光的MS细胞。在较高放大倍数下,通过叠加显示可识别同一分泌颗粒上罗丹明和异硫氰酸荧光素的共存情况。这一发现通过免疫电子显微镜得到了证实。CLSM技术可能对MS细胞的研究有用。