The Endocrine Pancreas of African Lungfish: Light and Electron Microscopic Immunocytochemistry and Morphology

1991 ◽

Vol 49 ◽

pp. 272-273

Author(s):

Veronika Burmeister ◽

N. Ludvig ◽

P.C. Jobe

Keyword(s):

Microscopic Examination ◽

Free Electron ◽

Electron Microscopic Examination ◽

Ultrastructural Localization ◽

Rabbit Serum ◽

Electron Microscopic ◽

Electron Microscopic Immunocytochemistry ◽

Phosphate Buffered Saline ◽

Rat Brain Tissue ◽

Immune Rabbit

Electron microscopic immunocytochemistry provides an important tool to determine the ultrastructural distribution of various molecules in both normal and pathologic tissues. However, the specific immunostaining may be obscured by artifactual immunoreaction product, misleading the investigator. Previous observations show that shortening the incubation period with the primary antibody from the generally used 12-24 hours to 1 hour substantially reduces the artifactual immunostaining. We now extend this finding by the demonstration of artifact-free ultrastructural localization of the Ca2/calmodulindependent cyclic nucleotide phosphodiesterase (CaM-dependent PDE) immunoreactivity in brain.Anesthetized rats were perfused transcardially with phosphate-buffered saline followed by a fixative containing paraformaldehyde (4%) and glutaraldehyde (0.25%) in PBS. The brains were removed, and 40μm sections were cut with a vibratome. The sections were processed for immunocytochemistry as described by Ludvig et al. Both non-immune rabbit serum and specific CaM-dependent PDE antibodies were used. In both experiments incubations were at one hour and overnight. The immunostained sections were processed for electron microscopic examination.

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Ultrastructural Immunocytochemistry of Five Rat Pituitary Adenomas

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s1431927600003147 ◽

1980 ◽

Vol 38 ◽

pp. 724-725

Author(s):

D. J. McComb ◽

N. Ryan ◽

E. Horvath ◽

K. Kovacs ◽

E. Nagy ◽

...

Keyword(s):

Pituitary Adenomas ◽

Pituitary Tumors ◽

Electron Microscopic ◽

Electron Microscopic Immunocytochemistry ◽

Transmission Electron ◽

Rat Pituitary ◽

Radiation Induced ◽

Group 5 ◽

Group 2 ◽

Microscopic Techniques

Conventional light and electron microscopic techniques failed to clarify the cellular composition and derivation of spontaneous and induced, intrasellar and transplanted pituitary adenomas in rats (1). In the present work, electron microscopic immunocytochemistry was applied to evaluate five adenohypo-physial tumors using a technique described by Moriarty and Garner (2). Spontaneously occurring pituitary adenomas (group 1) were harvested from aging female Long-Evans rats. R-Amsterdam rats were treated with 2 x 1.0 mg estrone acetate (HogivaI) s.c. weekly for 6 months. Pituitary adenomas in excess of 30 mg were removed from these animals to make up the tumors of group 2. Groups 3 and 4 consisted of estrogen-induced autonomous transplan¬ted pituitary tumors MtT.WlO and MtT.F4. Group 5 was a radiation-induced transplanted autonomous pituitary tumor MtT.W5. The tumors of groups 3,4 and 5 were allowed to proliferate in host rats 6-8 weeks prior to removal for processing. Tissue was processed for transmission electron microscopy (glutaraldehyde fixation, OsO4 postfixation and epoxy resin embedding), and electron microscopic immunocytochemistry (3% paraformaldehyde fixation and Araldite embedding).

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