Ultrasound-amplified immunohistochemistry.

Podkletnova, I.; Alho, H

doi:10.1177/41.1.8417112

Cited by 19 publications

(8 citation statements)

References 16 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…To enhance antigen penetration, an ultrasound-amplified immunostaining method was used (54). Briefly, after incubation of the samples with normal blocking serum, ultrasound irradiation was carried out with the primary antiserum (1:3,000 in PBS-1% normal goat serum) in an ultrasound bath for 20 s, thereafter incubated with the same solution overnight at 4°C, and further processed for antigen visualization as described above.…”

Section: Methodsmentioning

confidence: 99%

Tissue-specific expression of the diazepam-binding inhibitor in Drosophila melanogaster: cloning, structure, and localization of the gene.

Kolmer¹,

Roos²,

Tirronen³

et al. 1994

Mol. Cell. Biol.

Self Cite

View full text Add to dashboard Cite

The diazepam-binding inhibitor (DBI; also called acyl coenzyme A-binding protein or endozepine) is a 10-kDa polypeptide found in organisms ranging from yeasts to mammals. It has been shown that DBI and its processing products are involved in various specific biological processes such as GABAA/benzodiazepine receptor modulation, acyl coenzyme A metabolism, steroidogenesis, and insulin secretion. We have cloned and sequenced the Drosophila melanogaster gene and cDNA encoding DBI. The Drosophila DBI gene encodes a protein of 86 amino acids that shows 51 to 56% identity with previously known DBI proteins. The gene is composed of one noncoding 5' and two coding exons and is localized on the chromosomal map at position 65E. Several transcription initiation sites were detected by RNase protection and primer extension experiments.Computer analysis of the promoter region revealed features typical of housekeeping genes, such as the lack of TATA and CCAAT elements. However, in its low GC content and lack of a CpG island, the region resembles promoters of tissue-specific genes. Northern (RNA) analysis revealed that the expression of the DBI gene occurred from the larval stage onwards throughout the adult stage. In adult flies, DBI mRNA and immunoreactivity were detected in the cardia, part of the Malpighian tubules, the fat body, and gametes of both sexes. Developmentally regulated expression, disappearing during metamorphosis, was detected in the larval and pupal brains. No expression was detected in the adult nervous system. On the basis of the expression of DBI in some but not all tissues with high energy consumption, we propose that in D. melanogaster, DBI is involved in energy metabolism in a manner that depends on the substrate used for energy production.The diazepam-binding inhibitor (DBI) is a 10-kDa polypeptide that was first purified from rat brain on the basis of its ability to displace diazepam from the -y-aminobutyric acid receptor type A (GABAA)/benzodiazepine receptor (24). More recently DBI has also been purified from several peripheral organs such as pig intestine (14) and from bovine and rat liver (34,44,47). DBI has been shown to be expressed in several rat tissues (3, 9). The highest expression has been observed in the adrenal gland, liver, and somatic tissue of the testes, while the lowest levels have been detected in spleen, lung, and muscle (34,45,68). Nevertheless, DBI expression is restricted to certain cell types. In the rat brain, DBI is localized in selected neuronal populations, ependymal and glial cells (2, 3). DBIlike peptides have also been found in the central nervous systems of trout (Salmo gairdneri) (39) and frogs (Rana ridibunda) (40). In the adrenal gland, DBI is expressed in cortical cells, whereas chromaffin cells of the medulla are immunonegative (9). DBI and its metabolites octadecaneuropeptide (DBI33_50) and triakontatetraneuropeptide (DBI17-50) are involved in the regulation of multiple biological processes. In rats, intracerebroventricular injection of DBI or one of its metabolit...

show abstract

Section: Methodsmentioning

confidence: 99%

Tissue-specific expression of the diazepam-binding inhibitor in Drosophila melanogaster: cloning, structure, and localization of the gene.

Kolmer¹,

Roos²,

Tirronen³

et al. 1994

Mol. Cell. Biol.

Self Cite

View full text Add to dashboard Cite

show abstract

“…16 Since the 1960s, a few groups have reported using ultrasound to increase the speed of fixation 17 and processing 18,19 and to accelerate immunoassay 20,21 and staining. 22 Most of these studies used commercially available ultrasound generators and involved the use of low-frequency ultrasound (approximately 40 kHz), which led to controversial results and frequent tissue damage.…”

mentioning

confidence: 99%

Ultrasound-accelerated formalin fixation of tissue improves morphology, antigen and mRNA preservation

et al. 2005

View full text Add to dashboard Cite

Formalin fixation and paraffin embedding are conventional tissue preservation and processing methods used for histologic diagnosis in over 90% of cases. However, formalin fixation has three disadvantages: (1) slow fixation (16-24 h) hinders intraoperative decision making, (2) slow quenching of enzymatic activity causes RNA degradation, and (3) extensive molecule modification affects protein antigenicity. Applying high-frequency, high-intensity ultrasound to the formalin fixative cuts fixation time to 5-15 min. Fixation of various tissues such as lymph node, brain, breast, and prostate suggests that, compared to the conventional method, implementation of ultrasound retains superior and more uniform tissue morphology preservation. Less protein antigenicity is altered so that rapid immunohistochemical reactions occur with higher sensitivity and intensity, reducing the need for antigen retrieval pretreatment. Better RNA preservation results in stronger signals in in situ hybridization and longer RNA fragments extracted from fixed tissues, probably due to rapid inhibition of endogenous RNase activity. Molecules extracted from ultrasound-fixed tissues are of greater integrity and quantity compared to conventionally fixed tissues, and thus better support downstream molecular analyses. Overall, ultrasound-facilitated tissue preservation can provide rapid and improved morphological and molecular preservation to better accommodate both traditional and molecular diagnoses.

show abstract

“…9,31 The physical process of opening holes or channels within the tissue section also likely explains the modest success of ultrasonics as an AR method. 92 Successful application of heat -induced AR requires regulation of the pH of the recovery buffer. Shi et al 31,93 found three patterns of pH dependence as judged by immunostaining: (1) antigen recovery is independent of pH (stable type), (2) antigen recovery improves with increasing pH (ascending type), and (3) optimal antigen recovery occurs at low or high pH (V -type).…”

Section: General Comments On the Mechanism Of Armentioning

confidence: 99%