摘要:

The routine use of the micronucleus test in the mutagenicity evaluation of xe-nobiotics is limited by high cost and limited availability of fetal calf serum. On the other hand, there are disadvantages, such as hypotonic damage and clumping of cells, associated with the use of mineral medium substitutes for fetal calf serum. Alternatively, we recommend a chemical medium containing Hanks' buffered salt solution,1%(w/v) bovine serum albumin, and 0.15% (w/v) ethylenediaminetetraacetic acid, final pH 7.4, to preserve morphology, density and homogeneity of bone marrow cells. Mast cell granules are efficiently removed from rat bone marrow preparations by washing twice with this medium. The morphological preservation of cells is further enhanced by fixation with 70 % (v/v) ethanol for 5 min. The proposed medium provides a cost-effective and convenient substitute for fetal calf serum with substantially improved quality of bone marrow preparations for the micronucleus test.

ISSN:1052-0295    

DOI:10.3109/10520299309104695

出版商:Taylor&Francis    

年代:1993

数据来源: Taylor

摘要:

Hydrophobic adhesive tape was used to produce miniature wells on microscope slides for staining several sections of tissue with minimal amounts of cytochemical reagents. The wells could be tailored to individual specifications and the method allowed coverslips to be mounted close to the sections using either aqueous or xylene based mounting media. This method was especially useful for multiple immunolabelling of serial semithin cryosections.

ISSN:1052-0295    

DOI:10.3109/10520299309104696

出版商:Taylor&Francis    

年代:1993

数据来源: Taylor

摘要:

The demonstration of prophenoloxidase after electrophoresis is based on its activation by sodium dodecyl sulfate (SDS) or sodium oleate and staining the activated phenoloxidase with dopamine and 3-methyl-2-benzothiazolinone hydrazone hydrochloride (MBTH). A rapid method is presented for demonstrating the presence of activated phenoloxidase using polyacrylamide gel electrophoresis followed by staining in the presence of SDS or sodium oleate.

ISSN:1052-0295    

DOI:10.3109/10520299309104697

出版商:Taylor&Francis    

年代:1993

数据来源: Taylor

摘要:

Separation of the endocrine from the exocrine pancreatic tissue by fluorescence activated sorting has been limited by the lack of an ideal fluorescent label for islet tissue. Our studies indicates the zinc-specific stain N-(6-methoxy-8-quinolyl)-para-toluenesulfonamide (TSQ), has characteristics ideal for use as a fluorescent label for islet tissue. Dispersed rat pancreas cells stained with TSQ produced bright blue fluorescence when excited by UV light [peak emission wavelength at 480 nm. maximal excitation at 365 nm). The fluorescence was specific for islet tissue as confirmed by counterstaining with the islet-specific stain dithizone and there was minimal background staining of exocrine tissue. Stained tissue remained brightly fluorescent for 2 hr. with some fading by 4 hr. Injection of TSQ into rats at a concentration sufficient to produce staining of islets produced no toxicity discernible at 4 months. The viability of isolated rat islets stained with TSQ was maintained as shown by supravital staining, in vitro secretion of insulin, and reversal of diabetes after transplantation of stained islets into diabetic syngeneic recipients.

ISSN:1052-0295    

DOI:10.3109/10520299309104698

出版商:Taylor&Francis    

年代:1993

数据来源: Taylor

摘要:

An incubation protocol to immunolabel Lowicryl semithin sections was applied to paraffin probes. To improve the labeling density, colloidal gold complexes of different preparations and sizes were compared. The type of colloidal gold preparation used was found to affect the specificity of the immunostaining. Gold colloid of 5 nm diameter particle size prepared with white phosphorus minimized nonspecific background labeling ofβ-casein in paraffin embedded sections of the mammary epithelium of pregnant mice. Gold colloids of 5 nm and 9 nm diameter particle size prepared in varying concentrations of tannic acid generated significant nonspecific staining in similar tissue preparations.

ISSN:1052-0295    

DOI:10.3109/10520299309104699

出版商:Taylor&Francis    

年代:1993

数据来源: Taylor

摘要:

The freeze substitution device described here consists of commonly available materials, is easy to construct and to handle and provides maximal working safety. An adequate mass of copper and/or brass, serving as holder for 16 cryovials. is chilled to liquid nitrogen temperature. The vial holder is placed in a precooled thermos flask filled with ethanol and stored in an ultrafreezer at 193 K/-90 C. This ensures that the vial holder maintains temperatures of about 183 K/-90 C for more than 12 hr to minimize the risk of ice recrystallization.

ISSN:1052-0295    

DOI:10.3109/10520299309104700

出版商:Taylor&Francis    

年代:1993

数据来源: Taylor

摘要:

A new permanent staining procedure for sister chromatid differentiation (SCD) in cultured Chinese hamster ovary cells has been established by combining the three-way differentiation in third mitosis (M3) chromosomes and the immunoperoxidase reaction developed with 3,3′-diaminobenzidine using a monoclonal anti-bromodeoxyuridine (BrdU) antibody. This procedure allows SCD at very low BrdU concentrations, and the evaluation of the sister chromatid exchange frequencies on a per cell-cycle basis.

ISSN:1052-0295    

DOI:10.3109/10520299309104701

出版商:Taylor&Francis    

年代:1993

数据来源: Taylor

摘要:

Modified protocols for cobalt-filling and silver intensification of neurons in the larval and adult stages of the moth,Manduca sexta, have led to improved neuronal visualization and minimal background staining. In particular, long distance projecting multisegmental in-terneurons. originating in the pterothoracic or terminal abdominal ganglion, were best visualized when a cobalt:lysine complex was used to fill hemi-connectives for several days at 4 C. Ganglia closest to the placement of tracer, which became flooded with cobalt:lysine during the filling period. were removed from the insect. This step eliminated the artifactual filling of neurons that may have taken up the tracer from such pooled regions. This led to a more accurate assessment of whether a multisegmental interneuron projected through the full length of nerve cord to the original site of tracer placement. The protocol for light insensitive silver intensification of cobalt-filled neurons was modified to include an important pH adjustment. NaOH was used to alter the pH of the protective colloid, sodium tungstate, to 10.4 or greater in solution. Especially in larvae. our techniques produced intensely stained cobalt-filled neurons within ganglia that remained transparent and relatively free of nonspecific silver deposition.

ISSN:1052-0295    

DOI:10.3109/10520299309104702

出版商:Taylor&Francis    

年代:1993

数据来源: Taylor

摘要:

When investigating microscopic preparations perfusion chambers allow exchange and regulation of different solutions and ensure their constant flow in the sample chamber. Temperature deviations, however, may be problematic. We describe a new chamber that contains an additional circulation system which regulates the inside temperature using an external thermostat. An integrated thermometer probe records the sample temperature, which appears on a monitor. The glass chamber. measuring 75×35×3 mm, provides good optical quality and is compatible with every type of microscope.

ISSN:1052-0295    

DOI:10.3109/10520299309104703

出版商:Taylor&Francis    

年代:1993

数据来源: Taylor

摘要:

We have developed a double staining method using 5-bromo,4-chloro-3-indolyl-β-D-galactopyranoside X-gal and immunoperoxidase for wholeDrosophilaembryos. The dechorionated embryos are fixed in heptane saturated with 4% formaldehyde, then in heptane and 50% methanol. Fixed embryos are devitellinized with a tungsten needle and processed for immunoperoxidase staining immediately prior to peroxidase color development. The embryos are stained with X-gal, then peroxidase staining is resumed. This procedure enables us to observe cells stained with both X-gal and a specific antibody in whole embryos.

ISSN:1052-0295    

DOI:10.3109/10520299309104704

出版商:Taylor&Francis    

年代:1993

数据来源: Taylor