scholarly journals Triple immunohistochemical staining for bromodeoxyuridine and catecholamine biosynthetic enzymes using microwave antigen retrieval.

1995 ◽  
Vol 43 (1) ◽  
pp. 1-4 ◽  
Author(s):  
A S Tischler
Keyword(s):  
Chromaffin Cells ◽  
Antigen Retrieval ◽  
Adrenal Chromaffin Cells ◽  
Paraffin Sections ◽  
Efficient Tool ◽  
Formalin Fixed Paraffin ◽  
Adrenal Chromaffin ◽  
Formalin Fixed ◽  
Better Than

A method was developed for detection of bromodeoxyuridine (BrdU) in conjunction with other antigens in formalin-fixed paraffin sections with microwave antigen retrieval. The method was applied to rat adrenal medulla to demonstrate S-phase nuclei in epinephrine-producing cells stained for immunoreactive phenylethanolamine-N-methyltransferase and in norepinephrine-producing cells stained for immunoreactive tyrosine hydroxylase. The quality of staining for all three antigens was comparable to or better than that previously obtained with other techniques. This method provides an efficient tool for studying turnover of subpopulations of adrenal chromaffin cells. It should also be widely applicable to other cells and tissues.

scholarly journals Antigen retrieval technique utilizing citrate buffer or urea solution for immunohistochemical demonstration of androgen receptor in formalin-fixed paraffin sections.

1993 ◽  
Vol 41 (11) ◽  
pp. 1599-1604 ◽  
Author(s):  
S R Shi ◽  
B Chaiwun ◽  
L Young ◽  
R J Cote ◽  
C R Taylor
Keyword(s):  
Androgen Receptor ◽  
Enzymatic Digestion ◽  
Antigen Retrieval ◽  
Urea Solution ◽  
Paraffin Sections ◽  
Buffer Solution ◽  
Solution Ph ◽  
Citrate Buffer ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed

We developed a staining protocol for demonstration of androgen receptor (AR) in formalin-fixed, paraffin-embedded tissue sections. The method is based on the antigen retrieval microwave (MW) heating technique. Results are compared with different types of enzyme digestion pre-treatments. The strongest immunostaining signal and clearest background were obtained by MW heating of dewaxed paraffin sections in 5% urea or citrate buffer solution (pH 6); pure distilled water gave less consistent results. Enzymatic digestion with pepsin (0.05% in 2 N HCl) for 30 min at room temperature, or trypsin followed by pronase, or pronase digestion alone, also produced enhanced staining of AR in some cases, but there was more nonspecific background, and specific reactivity was less intense. The antigen retrieval MW method can be used to demonstrate AR epitope in prostate tissue after fixation in formalin for as long as 7 days. AR immunolocalization was also compared in frozen and paraffin sections processed from the same specimen of prostate carcinoma tissue and was found to be qualitatively and quantitatively similar. This study also provided new information concerning the basic principles of the antigen retrieval MW method that may be helpful in further development of this technique.

scholarly journals Epitopes of Components of the Plasminogen Activation System are Re-exposed in Formalin-fixed Paraffin Sections by Different Retrieval Techniques

1998 ◽  
Vol 46 (4) ◽  
pp. 469-476 ◽  
Author(s):  
Cilia M. Ferrier ◽  
Winny L. van Geloof ◽  
Hans H. de Witte ◽  
Michael D. Kramer ◽  
Dirk J. Ruiter ◽  
...  
Keyword(s):  
Negative Control ◽  
Antigen Retrieval ◽  
Plasminogen Activation ◽  
Paraffin Sections ◽  
Systematic Analysis ◽  
Plasminogen Activation System ◽  
Formalin Fixed Paraffin ◽  
Activation System ◽  
Formalin Fixed ◽  
Pai 1

We present a systematic analysis of the sensitivity and specificity of immunohistochemical stainings for components of the plasminogen activation system, i.e., uPA, tPA, PAI-1, PAI-2, and uPAR, on routinely processed (formalin-fixed, paraffin-embedded) tissues. Five to nine antibodies per component were tested and the influence of different antigen retrieval regimens on immunoreactivity was investigated. We studied six different microwave-mediated pretreatments and two pretreatments by proteolytic digestion. First, positive and negative control tissues were stained. Then, frozen and paraffin sections from the same cancer lesions were stained after specific modes of pretreatment and with selected antibodies. For each component, one or a few of the tested Abs gave optimal staining on paraffin sections when combined with a particular tissue pretreatment. For PAI-1, and to a lesser degree also for tPA, an underrepresentation of stromal cell staining in paraffin material was found, whereas tumor cells showed good staining. For uPA, PAI-2, and uPAR, consistent staining results were obtained on paraffin sections.

scholarly journals Immunostaining of chain-specific keratins on formalin-fixed, paraffin-embedded tissues: a comparison of various antigen retrieval systems using microwave heating and proteolytic pre-treatments.

1995 ◽  
Vol 43 (4) ◽  
pp. 429-437 ◽  
Author(s):  
H M Hazelbag ◽  
L J van den Broek ◽  
E B van Dorst ◽  
G J Offerhaus ◽  
G J Fleuren ◽  
...  
Keyword(s):  
Microwave Heating ◽  
Proteolytic Enzymes ◽  
Antigen Retrieval ◽  
Detergent Solution ◽  
Immunoperoxidase Method ◽  
Paraffin Sections ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Pre Treatment ◽  
Formalin Fixed

The use of chain-specific monoclonal antibodies (MAbs) against keratins in pathology is hampered by their limited staining on formalin-fixed, paraffin-embedded tissue. In the present study, various treatments before immunohistochemistry on paraffin sections were compared, including proteolytic enzymes and microwave antigen retrieval in various solutions. Sections of normal cervical and skin tissue were stained in a three-step immunoperoxidase method, employing a broad panel of MAbs against chain-specific keratins 4, 5, 7, 8, 10, 13, 14, 17, 18, 19 and pankeratin. Using microwave heating, Target Unmasking Fluid (TUF), Antigen Retrieval Solution (ARS), a simple detergent solution (DET), PBS, and distilled water (MiQ) were compared. Microwave heating in PBS or MiQ strongly improved staining results. Moreover, microwave pre-treatment in TUF or DET gave excellent and specific staining with the majority of MAbs tested, comparable with or even better than staining obtained on frozen sections. Using microwave antigen retrieval, tissue morphology remained optimal, and only in a very limited number of MAbs did immunoreactivity on paraffin sections fail to be restored. Proteolytic pre-treatment with trypsin, pepsin, or pronase gave moderate to strong staining with some of the MAbs. Other MAbs, for which microwave pre-treatment was able to restore the loss of immunoreactivity, failed to give appropriate staining with proteolytic pre-treatment. Our results show that microwave heating in either TUF or a simple detergent solution before immunohistochemistry is a reliable method for antigen retrieval of chain-specific keratins in formalin-fixed, paraffin-embedded tissues.

Morphological Observations on the Granule Types in Rabbit Extra-Adrenal Chromaffin Cells.

1975 ◽  
Vol 33 ◽  
pp. 318-319
Author(s):  
Joe A. Mascorro ◽  
Robert D. Yates
Keyword(s):  
Chromaffin Cells ◽  
Sodium Phosphate ◽  
Ganglion Cells ◽  
Ultrastructural Level ◽  
Osmic Acid ◽  
Adrenal Chromaffin Cells ◽  
Potassium Iodate ◽  
Perfusion Fluid ◽  
New Zealand White Rabbits ◽  
Adrenal Chromaffin

Extra-adrenal chromaffin organs (abdominal paraganglia) constitute rich sources of catecholamines. It is believed that these bodies contain norepinephrine exclusively. However, the present workers recently observed epinephrine type granules in para- ganglion cells. This report investigates catecholamine containing granules in rabbit paraganglia at the ultrastructural level.New Zealand white rabbits (150-170 grams) were anesthetized with 50 mg/kg Nembutal (IP) and perfused with 3% glutaraldehyde buffered with 0.2M sodium phosphate, pH 7.3. The retroperitoneal tissue blocks were removed and placed in perfusion fluid for 4 hours. The abdominal paraganglia were dissected from the blocks, diced, washed in phosphate buffer and fixed in 1% osmic acid buffered with phosphate. In other animals, the glutaraldehyde perfused tissue blocks were immersed for 1 hour in 3% glutaraldehyde/2.5% potassium iodate buffered as before. The paraganglia were then diced, separated into two vials and washed in the buffer. A portion of this tissue received osmic acid fixation.

scholarly journals Stimulatory effect of serotonin on Na+-dependent Ca2+ efflux from bovine adrenal chromaffin cells in culture

1997 ◽  
Vol 73 ◽  
pp. 226
Author(s):  
Kazuo Minakuchi ◽  
Hitoshi Houchi ◽  
Masanori Yoshizumi ◽  
Yasuko Ishimura ◽  
Kyoji Morita ◽  
...  
Keyword(s):  
Chromaffin Cells ◽  
Adrenal Chromaffin Cells ◽  
Cells In Culture ◽  
Adrenal Chromaffin
Sign in / Sign up

Export Citation Format

Share Document