ISSN:1052-0295    

DOI:10.3109/10520299409106294

出版商:Taylor&Francis    

年代:1994

数据来源: Taylor

摘要:

Slides pretreated for C-banding and stained with DAPI or CMA3 show different banding patterns in human metaphase chromosomes compared to those obtained with either standard Giemsa C-banding or fluorochrome staining alone. Human chromosomes show C-plus DA-DAPI banding after C-banding plus DAPI and enhanced R-banding after C-banding plus Chromomycin A3 staining. If C-banding preferentially removes certain classes of DNA and proteins from different chromosome domains, C-banding pre-treatment may cause a differential DNA extraction from G- and R-bands in human chromosomes, resulting in a preferential extraction of DNA included in G-bands. This hypothesis is partially supported by the selective cleavage and removal of DNA from R-bands of restriction endonuclease HaeIII with C-banding combined with DAPI or Chromomycin A3 staining. Structural factors relating to regional differences in DNA and/or proteins could also explain these results.

ISSN:1052-0295    

DOI:10.3109/10520299409106295

出版商:Taylor&Francis    

年代:1994

数据来源: Taylor

摘要:

A nonisotopic, double fluorescence technique was developed to study myogenic satellite cell proliferation in posthatch turkey skeletal muscle. Labeled satellite cell nuclei were identified on enzymatically isolated myofiber segments using a mouse monoclonal antibody (anti-BrdU) followed by fluorescein-5-isothiocyanate (FITC) conjugated goat anti-mouse IgG secondary antibody. Myofiber nuclei (myonuclei + satellite cell nuclei) were counterstained with propidium iodide (PI). The myofiber segment length, myofiber segment diameter, and the number of PI and FITC labeled nuclei contained in each segment was determined using a Nikon fluorescence microscope, a SIT video camera and Image-1 software. Data collected by three different operators of the image analysis system revealed 5.0±1.4 satellite cell nuclei per 1000 myofiber nuclei and 5284±462μm3of cytoplasm surrounding each myofiber nucleus in the pectoralis thoracicus of 9-week-old tom turkeys. BrdU immunohistochemistry coupled with the new approach of PI staining of whole myofiber mounts is an effective combination to allow the use of an efficient semi-automated image analysis protocol.

ISSN:1052-0295    

DOI:10.3109/10520299409106296

出版商:Taylor&Francis    

年代:1994

数据来源: Taylor

摘要:

A device for cutting brain slices is described as an alternative to cutting angle guides and the“brain macrotome”. With this new device, slices of uniform thickness optimal for assessing morphological detail and photography can be produced. A similar but smaller device for cutting pieces of tissue for paraffin embedding is also presented. These devices should be useful in either the histopathology laboratory or mortuary.

ISSN:1052-0295    

DOI:10.3109/10520299409106297

出版商:Taylor&Francis    

年代:1994

数据来源: Taylor

摘要:

Mineralization of renal tissue was investigated using Von Kossa's method and capillary gap technology. Combining these techniques allowed the simultaneous processing of 100 slides, reduced the volume of 5% silver nitrate required, and produced consistent dark brown to black silver deposits in positive control sections. A 25-fold savings in silver nitrate was achieved. By markedly reducing individual slide handling, technician time also was greatly reduced. Two or three slide sets, each containing approximately 100 slides, can be conveniently stained daily.

ISSN:1052-0295    

DOI:10.3109/10520299409106298

出版商:Taylor&Francis    

年代:1994

数据来源: Taylor

摘要:

Ethyl N-methyl-4-hydroxy-5-oxo-3-pyrroline-3-carboxylate forms a deep red chelate with iron salts. The color intensity is directly related to the iron concentration. The photosta-bility of the red color was determined at pH 1.2 and 5 by spectrophotometric assay at 484 nm at intervals during irradiation by tungsten light at 1020μW/cm2. After 528 hr of continuous irradiation in deionized water, 90.9% of the iron chelate had decomposed. The reaction followed zero order kinetics. Maximal stability was observed at pH 5 at both 10--2and 10--2molar concentrations of the iron chelate: no detectable decomposition occurred after 192 hr of continuous irradiation. The iron chelate in biological tissues is stable for 18 months. The staining technique is superior to other histological methods for estimating low concentrations of iron in tissue.

ISSN:1052-0295    

DOI:10.3109/10520299409106299

出版商:Taylor&Francis    

年代:1994

数据来源: Taylor

摘要:

Although well established myocar-dial infarcts (MI) are not difficult to identify in routine hematoxylin and eosin stained sections. recent MI may present diagnostic problems. Fibrinogen is a useful marker for detecting early ischemic cell damage. Using immunofluorescence on frozen tissue. albumin, IgG and fibrinogen have been found throughout the sarcoplasm of ischemic fibers in human hearts. In this report, monoclonal antibodies to all three proteins were reacted using the avidin-biotin technique in formalin fixed, paraffin embedded sections from autopsy cases of sudden or intraoperative deaths with either subtle or no definite ischemic changes evident in routine sections. Strong staining of fibrinogen in the fibers associated with coagula-tive necrosis, contraction bands or wavy fibers, and in the fibers presumably associated with acute ischemia. Albumin and IgG staining was nonspecific. Fibrinogen is a reliable and reproducible marker for recognizing early ischemia. This method can be used to diagnose early sudden ischemic and intraoperative deaths due to coronary artery bypass graft and prosthesis-related complications and may be particularly useful for forensic autopsies.

ISSN:1052-0295    

DOI:10.3109/10520299409106300

出版商:Taylor&Francis    

年代:1994

数据来源: Taylor

摘要:

Techniques for impregnation with ammoniacal silver carbonate provide valuable information on all types of tissue; however, the time investment required to impregnate a few sections has limited their application. We have shortened the impregnation times by using microwaves in techniques for reticular fibers, astrocytes, nerve fibers and chromaffin cells. The results were satisfactory with markedly reduced impregnation time and elimination of nonspecific silver deposits.

ISSN:1052-0295    

DOI:10.3109/10520299409106301

出版商:Taylor&Francis    

年代:1994

数据来源: Taylor

摘要:

Since its introduction in 1971, the four-and-a-half clearing technique has been widely applied to the study of ovule and female gametophyte development in flowering plants as an alternative to the more arduous paraffin section methods. The technique has undergone several modifications that have broadened its application in studies of Angiosperm embryology. To date, however, the technique has not been successfully applied to embryological features of Gymnosperms. Dark coloration caused by naturally occurring substances and by-products of fixation render the clearing fluid ineffective, and special pretreatment methods used to remove dark substances in Angiosperm ovules have little or no effect on Gymnosperm material. In the technique reported here, paraffin sections of ovules and young seeds ofCunninghamia lanceolata80–120μm thick are cleared in benzyl benzoate-412 clearing fluid and examined with phase contrast optics. Observations of the mature female gametophyte in these cleared preparations are compared with those obtained from 10μmsections, stained with safranin and fast green, and examined with bright-field optics. Although contrast and definition are more pronounced in stained sections than in cleared ones, the differences would not alter one's interpretation of characteristic structural features. The thick, cleared section offers an advantage over the thin, stained one in that many structural entities are contained within a single section rather than spread through several serial sections. The time required for clearing thick sections is much shorter than that required for making permanent stained preparations.

ISSN:1052-0295    

DOI:10.3109/10520299409106302

出版商:Taylor&Francis    

年代:1994

数据来源: Taylor

摘要:

We have developed an improved assay for the production of collagen by fibroblasts. Early passage adult mouse lung fibroblasts, established and maintained in serum-free culture, were employed as the target cells. An enzyme immunoassay was used for detection of type I collagen deposited on the substratum, permitting adaptation of the technique to cultures in 96-well microplates. Approximately two-fold enhancement of collagen deposition was induced by exposure to a concentration of 3 ng/ml of transforming growth factor-β1 or of 100 ng/ml of insulin-like growth factor-1 for 48 hr.

ISSN:1052-0295    

DOI:10.3109/10520299409106303

出版商:Taylor&Francis    

年代:1994

数据来源: Taylor