Modified Thiocarbohydrazide Procedure for Scanning Electron Microscopy: Routine use for Normal, Pathological, or Experimental Tissues

Malick, Linda E.; Wilson, R. B.; Stetson, David L.

doi:10.3109/10520297509117069

Cited by 249 publications

(96 citation statements)

References 3 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Sections were washed in 0.1 M sodium cacodylate buffer three times, then incubated with a solution of 20 mM DTT, 150 mM Tris, and 20% ethanol for 45 min at room temperature to remove mucin. Sections were washed in 0.1 M sodium cacodylate buffer three times, treated with alternating 1% osmium tetroxide and 1% thiocarbohydrazide (OTOTO method) (22), dehydrated in graded series of ethanol, critical point dried, and sputter-coated to produce 15-nm gold coating. Samples were examined using a Hitachi S-450 scanning electron microscope (Hialeah, FL) at an accelerating voltage of 20 kV.…”

Section: Scanning Electron Micrographsmentioning

confidence: 99%

Isolated Lymphoid Follicle Formation Is Inducible and Dependent Upon Lymphotoxin-Sufficient B Lymphocytes, Lymphotoxin β Receptor, and TNF Receptor I Function

Lorenz

Chaplin

McDonald

et al. 2003

The Journal of Immunology

256

362

View full text Add to dashboard Cite

The gastrointestinal mucosa contains a complex network of lymphoid compartments that have evolved to efficiently protect the host from invading pathogens. Recently, an additional lymphoid structure resembling Peyer’s patches (PP) in composition and architecture has been identified in the murine small intestine, the isolated lymphoid follicle (ILF). In this study we examine the nature and factors required for ILF formation. We observed a spectrum of structures fitting the previous descriptions of ILFs, ranging from clusters of B220+ cells (which we have termed immature ILFs) to well-organized lymphoid nodules (which we have termed mature ILFs). Here we demonstrate that that similar to PP formation, ILF formation requires lymphotoxin (LT)- and LTβ receptor-dependent events. However unlike PP formation, the LT- and LTβ receptor-dependent events required for ILF formation can occur in adulthood and require LT-sufficient B lymphocytes. We demonstrate that mature ILF formation occurs in response to lumenal stimuli, including normal bacterial flora, and requires TNF receptor I function. These findings suggest that ILFs are organized intestinal lymphoid structures whose formation can be induced and whose mass can be expanded in response to mucosal challenges.

show abstract

Section: Scanning Electron Micrographsmentioning

confidence: 99%

Isolated Lymphoid Follicle Formation Is Inducible and Dependent Upon Lymphotoxin-Sufficient B Lymphocytes, Lymphotoxin β Receptor, and TNF Receptor I Function

Lorenz

Chaplin

McDonald

et al. 2003

The Journal of Immunology

256

362

View full text Add to dashboard Cite

show abstract

“…Microcarriers coated with cells were fixed for light and electron microscopy in 0.1 M cacodylate buffered 3% glutaraldehyde (pH 7.2) for at least 1 h. The microcarriers were prepared for scanning electron microscopy (Malick & Wilson, 1975) and examined on a Philips 500 SEM. The cells exhibited the 'cobblestone' morphology and numerous microvilli characteristic of endothelial cells (Figure 1).…”

Section: (B) Electron Microscopymentioning

confidence: 99%

Bioassay of prostacyclin and endothelium‐derived relaxing factor (EDRF) from porcine aortic endothelial cells

Gryglewski

Moncada

Palmer

1986

British J Pharmacology

217

View full text Add to dashboard Cite

A cascade superfusion technique has been developed for the differential bioassay of prostacyclin and endothelium-derived relaxing factor (EDRF) released from porcine aortic endothelial cells cultured on microcarriers, packed into a column and perfused. 2 Bradykinin (Bk; 20-100 nM) released prostacyclin (9.6 ± 1.5 nm per 106 cells; mean ± s.e.mean, n = 9) and prostaglandin E2 (PGE2; 2.1 ± 0.6 nm per 10' cells) from the column measured by relaxation of strips of bovine coronary artery (BCA) and rabbit mesenteric or coeliac artery, respectively. The presence of these prostanoids in the effluent was confirmed by specific radioimmunoassays. 3 A23187 (500-2000nM) also released both prostacyclin and PGE2 from the cells. This release was long-lasting and not reproducible. 4 Bk (20-100 nM) and A23187 (30-300 nM) released EDRF from the column. This was detected in a cascade of four rabbit aortic strips (RbA), denuded of endothelium and contracted with U46619 or phenylephrine. The relaxation of the RbA strips caused by EDRF was progressively attenuated down the cascade (half-life < 7 s) and was not affected by indomethacin. 5 EDRF and prostacyclin could be differentially bioassayed in a cascade of alternating RbAs and BCAs as prostacyclin did not relax RbAs and the time delay to the BCAs destroys EDRF. EDRF could be bioassayed on its own when the endothelial cells were treated with indomethacin. 6 5-Hydroxytryptamine 0.2, noradrenaline 1.0, platelet-activating factor (Paf-acether) 1.0, formylmethionyl-leucyl-phenylalanine 1.0, acetylcholine 0.5, bethanecol 0.5, adenosine diphosphate 0.25 and angiotensin II 0.1 gM did not release either prostanoids or EDRF from the column.

show abstract

“…Urethral plugs (UP) were harvested, washed in distilled water, dried with filter paper and weighed (Table I). Specimens for scanning electron microscopy were immersed in fixative (2'5% glutaraldehyde and 2% paraformaldehyde in 0'1 M cacodylate buffer, pH 7-3) for 24 h and further processed according to the O-T-O-T-O method (Malick, Wilson & Stetson, 1975), freeze-fractured, criticalpoint dried and 'gold-sputtered' for observation. Water content was determined from 50 pooled UP and contents of 50 pooled seminal vesicles after lyophilization; the nitrogen content was then determined by the micro-Kjeldahl method.…”

Section: Examinationmentioning

confidence: 99%

Urethral plug-a new secondary male sex characteristic in rat and other rodents

Kunstýr

Küpper²,

Weisser³

et al. 1982

Lab Anim

View full text Add to dashboard Cite

The plug is an eosinophilic mass, partly homogenous and partly porous, filling the proximal urethra in rats and occasionally extending into the bladder. Its average weight in 131 adult rats was 0·063 g. These plugs are normally present in the urethra of adult male rats, and this seems to be the case for all laboratory Muridae and Cavidae. The absence of a plug in an adult male may be a sign of abnormality associated with failing health. There is an interesting similarity between the amino acid composition of the content of seminal vesicles, that of the urethral plug, and that of the copulatory vaginal plug in female rodents.

show abstract

Modified Thiocarbohydrazide Procedure for Scanning Electron Microscopy: Routine use for Normal, Pathological, or Experimental Tissues

Cited by 249 publications

References 3 publications

Isolated Lymphoid Follicle Formation Is Inducible and Dependent Upon Lymphotoxin-Sufficient B Lymphocytes, Lymphotoxin β Receptor, and TNF Receptor I Function

Isolated Lymphoid Follicle Formation Is Inducible and Dependent Upon Lymphotoxin-Sufficient B Lymphocytes, Lymphotoxin β Receptor, and TNF Receptor I Function

Bioassay of prostacyclin and endothelium‐derived relaxing factor (EDRF) from porcine aortic endothelial cells

Urethral plug-a new secondary male sex characteristic in rat and other rodents

Contact Info

Product

Resources

About