摘要:

Immunohistochemical methods combined with progressive plasmolysis were used to localize chalcone synthase (CHS), an important enzyme for plant metabolism of aromatics in hypocotyls of illuminated buckwheat (Fagopyrum esculentum M) seedlings. Illumination of etiolated seedlings with white light results in anthocyanin synthesis in the epidermal layer of the hypocotyl. Anthocyanin-containing epidermal peels, after fixation for 30 min in 4% paraformaldehyde, 2.5% glutaraldehyde, 0.1% caffeine, were treated with a specific rabbit anti-buckwheat CHS antibody and a 20 nm goat anti-rabbit IgG gold conjugate. CHS is specifically shown in epidermal cells as pink to dark red deposits. Progressive plasmolysis combined with our immunohistochemical method showed that CHS was located exclusively in the cytoplasm of the epidermal cells of buckwheat hypocotyls except for the guard cells, which contained no detectable CHS.

ISSN:1052-0295    

DOI:10.3109/10520299509108308

出版商:Taylor&Francis    

年代:1995

数据来源: Taylor

摘要:

A simple method for light microscope screening of pre-embedded, gold-labeled samples, by silver enhancing deplasticized semithin epoxy sections is presented. This method is of great help in visualizing the topography of gold labeled sites at low magnification and for selecting labeled sample regions for subsequent electron microscope analysis on serial ultrathin sections. The method was developed during a study of lectin binding sites on the egg surface of the anuran amphibianDiscoglossus pictus

ISSN:1052-0295    

DOI:10.3109/10520299509108309

出版商:Taylor&Francis    

年代:1995

数据来源: Taylor

摘要:

Ethylenediaminetetraacetic acid (EDTA) solution is used to decalcify bone specimens for histological examination. Sodium hydroxide (NaOH) has been used to dissolve EDTA and to bring EDTA solutions to neutral pH. This solution, however, requires several weeks to decalcify bone specimens. We investigated a new de-calcification fluid using concentrated ammonium hydroxide (NH4OH) to dissolve EDTA and to adjust the pH to neutral. Decalcification was performed using a magnetic stirrer with and without vacuum, or with a sonic cleaner. Decalcification end point was confirmed using both the weight loss and X-ray methods. After decalcification, specimens were processed through paraffin and sections were stained with hematoxylin and eosin. Decalcification employing NH4OH required an average of six days. Light microscopy indicated good retention of cellular detail.

ISSN:1052-0295    

DOI:10.3109/10520299509108310

出版商:Taylor&Francis    

年代:1995

数据来源: Taylor

摘要:

A methanolic solution of the oxazine textile dye, C. I. basic blue 122, followed by an aqueous alkaline solution of the oxazine dye, C. I. basic blue 141, and a brief rinse in an acetate buffer at pH 3.45 produces intense black staining of eosinophil granules. This staining was selective for eosinophils while other types of peripheral blood leukocytes showed little if any staining under the same conditions. This staining procedure may be useful for detecting eosinophils in samples of blood, bone marrow, or urine when eosinophiluria results from interstitial nephritis.

ISSN:1052-0295    

DOI:10.3109/10520299509108311

出版商:Taylor&Francis    

年代:1995

数据来源: Taylor

摘要:

A quick embedding method using UV polymerization of methacrylate plastic has been devised for embedding fibers encased in a polyvinyl chloride tube. The resulting embedments are suitable for light microscopy and image analysis.

ISSN:1052-0295    

DOI:10.3109/10520299509108312

出版商:Taylor&Francis    

年代:1995

数据来源: Taylor

摘要:

Formalin fixed autopsy tissue containing lipids were cut into 1-5 nun thick blocks, washed well, then postfixed in 2% OsO4in 0.03 M veronal acetate buffer for 30, 60, 90, 120, or 180 min with or without ultrasonic treatment. Tissues exposed to ultrasound for 90 min showed superior penetration of OsO4and well preserved histological architecture. Tissues also were immersed for 1 hr in veronal acetate buffer (pH 7.4) containing 0.5% imidazole or triazole and compared with untreated controls. Paraffin sections, 4μm thick, were examined under a light microscope with an image analyzer. Both intensity and percentage area of osmium blackening were significantly higher in samples immersed in imidazole or triazole than in untreated controls. No difference was observed between imidazole- and triazole-immersed samples. The OsO4method, modified by ultrasound treatment and imidazole- or triazole-immersion, can be applied to routine formalin fixed autopsy materials for improved lipid visualization.

ISSN:1052-0295    

DOI:10.3109/10520299509108313

出版商:Taylor&Francis    

年代:1995

数据来源: Taylor

摘要:

A new histochemical technique, called in situ 3′-tailing reaction (ISTR), to detect DNA double strand breaks (DSB) was developed and applied to tissue sections of apoptotic endometrium. To demonstrate DSB, biotin-labeled and unlabeled dATPs with terminal deoxynucleotidyl transferase (TdT) were added to the many 3-hydroxyl termini of DNA fragments generated in the apoptotic cells. For an efficient 3′-end labeling, it was necessary to treat the sections withλ-exonuclease (λEx) prior to the TdT reaction to generate 3′-protruding ends. TheλEx-TdT reaction specifically labeled nuclear fragments in the apoptotic cells in paraformaldehyde fixed frozen sections. In paraffin sections, pretreatment with proteinase K was effective for 3′-tailing reaction. ISTR should be a useful tool for detecting dying cells in both physiological and pathological states.

ISSN:1052-0295    

DOI:10.3109/10520299509108314

出版商:Taylor&Francis    

年代:1995

数据来源: Taylor

摘要:

We evaluated the performance of four anti-fading agents during acquisition of multiple optical sections near the widest diameter ofDrosophilaaccessory gland nuclei using indirect immunofluorescence and confocal laser scanning microscopy. Two commercially available agents, Vectashield®and SlowFade®showed anti-fading properties that alleviated fluorochrome fading associated with the acquisition of multiple fluorescent optical Z-series from a single specimen by a confocal laser scanning system. Using these reagents, we were able to colocalize polypeptides through immunostained whole Drosophila nuclei.

ISSN:1052-0295    

DOI:10.3109/10520299509108315

出版商:Taylor&Francis    

年代:1995

数据来源: Taylor

摘要:

The effects of various handling procedures used in preparing specimens of human and pig respiratory mucosa for scanning electron microscopy (SEM) were studied. Using five different washing methods, the percentage area of mucosa covered with extracellular material varied from 1.5 to 53.1%. The best results were achieved when specimens were washed by gently inverting a sample 30 times in a container filled with physiological saline. Fixation and drying of the surface layer caused disorientation of cilia and made examination difficult. Mechanical damage caused loss of cilia and rupture of the cell membrane. For SEM of respiratory cilia it is important to wash the specimen in saline before fixation and to use biopsy forceps as little as possible.

ISSN:1052-0295    

DOI:10.3109/10520299509108316

出版商:Taylor&Francis    

年代:1995

数据来源: Taylor