摘要:

A procedure for fixing and immunostaining whole cells from primary cultures of ovine and bovine uterine gland fragments was used to identify keratin in intermediate filaments of epithelial cells to distinguish them from stromal cells. Colloidal gold encapsulated aga-rose-gelatin microbeads were coated with different proteins and used to investigate uptake by epithelial and stromal cells in culture. Micro-beads were taken up by stromal cells and by epithelial cells on the outskirts of colonies. These cells formed ridges where they contacted and grew above stromal cells. Electron microscopy demonstrated that the microbeads had been internalized and appeared to be nontoxic. Individual cells could harbor more than 90 microbeads within their cytoplasm for at least seven to ten days with no apparent harm. Some cells with microbeads were seen to divide.

ISSN:1052-0295    

DOI:10.3109/10520299109110540

出版商:Taylor&Francis    

年代:1991

数据来源: Taylor

摘要:

Six different staining techniques were evaluated for their suitability to stain nuclei ofColletotrichum gloeosporioidesf. sp.malvae(C.g.m.) spores. Of the three fluorescent stains, DAPI (4',6-diamidino-2-phenylindole) and bisbenzimide (Hoechst 33258) stained spore nuclei well; mithramycin did not. To achieve consistent results with the bisbenzimide staining protocol, the spores had to be fixed prior to staining and the stain had to be supplemented with Triton X-100. Both safranin O and Giemsa were suitable nonfluorescent staining techniques; lomofungin was not. Safranin O staining was simple and rapid. However, reproducibility was better if the spore suspension and KOH droplets were rapidly mixed prior to adding the stain. There was no significant difference in the percentages of uninucleate and binucleate spores observed in spore preparations stained with DAPI, bisbenzimide, safranin O or Giemsa. Bisbenzimide and safranin O were found to be simple, rapid and reliable fluorescent and nonfluorescent techniques, respectively, for staining nuclei of C.g.m. spores.

ISSN:1052-0295    

DOI:10.3109/10520299109110541

出版商:Taylor&Francis    

年代:1991

数据来源: Taylor

摘要:

It was necessary to make sections of small unfixed specimens which had been frozen while still immersed in their normal culture medium. The principal difficulty stemmed from the poor sectioning quality of the frozen culture medium. A capsule is described which has a narrow well in which the tissue specimen fits snugly within a small amount of culture medium. After freezing, the whole capsule is sectioned and the resulting sections, being nearly devoid of culture medium, are of good quality.

ISSN:1052-0295    

DOI:10.3109/10520299109110542

出版商:Taylor&Francis    

年代:1991

数据来源: Taylor

摘要:

A number of excellent techniques are available to stain and characterize different types of neurons and nerve terminals. However, because these different techniques are frequently not compatible, their usefulness in determining the relationships between specific axons and neuromuscular junctions is often limited. The goal was to develop specific procedures for simultaneous visualization of different types of unmyelinated axons and motor nerve terminals in the same preparation. First we modified the formal-dehyde/glutaraldehyde staining solutions of the aqueous aldehyde fluorescence technique (Faglu) to observe catecholamine containing axons in whole mount amphibian skeletal muscle. The compatibility of this modified staining solution with other histological procedures made it possible to stain both motor nerve terminals with tetrazolium salts and, in the same preparation, to observe unmyelinated axons with aldehyde-induced catecholamine histofluorescence. This same general formaldehyde/glutaraldehyde staining procedure was also used with immunocytochemical techniques to visualize fluorescent antibody stained nerves and motor nerve terminals in the same whole mount preparation.

ISSN:1052-0295    

DOI:10.3109/10520299109110543

出版商:Taylor&Francis    

年代:1991

数据来源: Taylor

摘要:

The e-amino groups of histone proteins were eliminated by condensation with glucose, fractose or mannose. The trichloroacetic acid extracted, DNA negative nuclei treated with reducing sugars, stained easily with basic dyes.

ISSN:1052-0295    

DOI:10.3109/10520299109110544

出版商:Taylor&Francis    

年代:1991

数据来源: Taylor

摘要:

A pH table is reported for citric acid-ammonium acetate buffers that are useful for horseradish peroxidase (HRP) histochemistry.

ISSN:1052-0295    

DOI:10.3109/10520299109110545

出版商:Taylor&Francis    

年代:1991

数据来源: Taylor

摘要:

The Hoechst dye staining method has been successfully applied to the central nervous system in mammals and its use has been demonstrated in intracerebral transplantation. The technique is rapid, simple and based on intrinsic nuclear properties. It was found to be permanent and valid whatever the animal strains or ages, allowing the distinction of rat cells from those of mouse, studied either separately or in a cross-transplantation model. It permitted the detection of grafted cells in the area of transplantation and the observation of early dispersion around the implantation site. Moreover, it can be combined with immunohistochemistry as demonstrated by a myelin marker in a relevant model. Immunodetection can thus help to directly observe grafted cells, at distance from the locus of transplantation, confirming their presence in the graft-type myelin patches.Because of its rapid performance, this technique can be used systematically after transplantation to check for the presence of grafted cells in the host.

ISSN:1052-0295    

DOI:10.3109/10520299109110546

出版商:Taylor&Francis    

年代:1991

数据来源: Taylor

摘要:

A staining procedure using celes-tine blue and chrome-trope 2R for Lowicryl K4M embedded tissues is presented. The stain produces a reliable multichromatic stain for light microscopy on Lowicryl embedded semithin sections.

ISSN:1052-0295    

DOI:10.3109/10520299109110547

出版商:Taylor&Francis    

年代:1991

数据来源: Taylor

摘要:

This paper addresses a practical problem associated with the use of visual detection systems used in immunoblotting. Western blot analyses from the same experiment, differing only at the level of the secondary antibody used and the means of visualization employed, have produced apparently different results which, in isolation, could lead to different conclusions at both the qualitative and quantitative level. Indirect immunofluorescence, using a fluorescein isothiocyanate labeled secondary antibody and visualized by fluorescence excitation, was excellent for detecting the major species present but could not detect minor components. Indirect immunoperoxidase staining, on the other hand, appeared to detect all immunoreactive species present, both major and minor, presumably reflecting a more realistic picture of the experimental situation. All results were obtained during the observation of photocross-link formation between regulatory light chains and regulatory and essential light chains after hybridization of scallop myosin with benzophenone-4-maleimide labeled regulatory light chains. Results were obtained under conditions designed to simulate the physiological states of rest and rigor; the implication of these results with respect to myosin-linked regulation is discussed.

ISSN:1052-0295    

DOI:10.3109/10520299109110548

出版商:Taylor&Francis    

年代:1991

数据来源: Taylor

摘要:

The employment of certain DNA-specific fluorescent stains on unhanded and C-banded chromosomes of two species of grasshoppers shows remarkable differences among C-heterochromatic regions supposed to be similar in their base pair composition, according to their response to the standard fluorescence techniques. The possible interspersion of the opposite DNA base pairs in these regions as well as the role played by proteins in chromosome banding are discussed.

ISSN:1052-0295    

DOI:10.3109/10520299109110549

出版商:Taylor&Francis    

年代:1991

数据来源: Taylor