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At least 19 recordsLinked to original sources
PubMed1978

Fixation variables in horseradish peroxidase neurohistochemistry. I. The effect of fixation time and perfusion procedures upon enzyme activity.

In a series of neurohistochemical experiments the effect of aldehyde fixation upon the detection of horseradish peroxidase (HRP) was examined. These experiments demonstrated that: a) Increments in fixation of as little as 1 hr significantly decreased the number of labeled neurons; 12-hr fixation abolished HRP activity in many neuronal populations and significantly reduced the apparent size of the injection site. b) This negative fixation effect was greatest where the HRP concentration was low (e.g. in small, lightly labeled neurons) but was still evident in areas of high concentration (e.g. large, heavily labeled neurons). c) This effect was also most prominent when a less sensitive diaminobenzidine histochemical procedure was employed but was still apparent with a more sensitive benzidine dihydrochloride procedure. d) Immersion of the brain in fixative after perfusion produced a greater attenuation of HRP activity in more superficial areas. e) Immersion of the brain in buffer to terminate fixation produced a prolonged and unpredictable gradient of fixation. f) Excess, unbound fixative inhibited the histochemical reaction per se and had to be removed from the tissue but prolonged washing did not resurrect enzyme activity which was lost by fixation. To obviate these problems and optimize HRP enzyme activity a new perfusion-fixation procedure was developed. It entails 30 min fixation by perfusion which is terminated by a subsequent 30 min perfusion with cold sucrose-fuller to wash out unbound fixative. This allows the tissue to be processed immediately, produces a uniform and morphologically adequate fixation, and minimizes the negative effects of fixation on HRP enzyme activity.

Animals↗
PubMed1975

The role of pH in the regulation of carbon fixation in the chloroplast stroma. Studies on CO2 fixation in the light and dark.

1. The pH in the stroma and in the thylakoid space has been measured in a number of chloroplast preparations in the dark and in the light at 20 degrees C. Illumination causes a decrease of the pH in the thylakoid space by 1.5 and an increase of the pH in the stroma by almost 1 pH unit. 2. CO2 fixation is shown to be strongly dependent on the pH in the stroma. The pH optimum was 8.1, with almost zero activity below pH 7.3.Phosphoglycerate reduction, which is a partial reaction of CO2 fixation, shows very little pH dependency. 3. Low concentrations of the uncoupler m-chlorocarbonylcyanide phenylhydrazone (CCCP) inhibit CO2 fixation without affecting phosphoglycerate reduction. This inhibition of CO2 fixation appears to be caused by reversal of light induced alkalisation in the stroma by CCCP. 4. Methylamine has a very different effect compared to CCCP. Increasing concentrations of methylamine inhibit CO2 fixation and phosphoglycerate reduction to the same extent. The light induced alkalisation of the stroma appears not to be significantly inhibited by methylamine, but the protons in the thylakoid space are neutralized. The inhibition of CO2 fixation by higher concentrations of methylamine is explained by an inhibition of photophosphorylation. It appears that methylamine does not abolish proton transport. 5. It is shown that intact chloroplasts are able to fix CO2 in the dark, yielding 3-phosphoglycerate. This requires the addition of dihydroxyacetone phosphate as precursor of ribulosemonophosphate and also to supply ATP, and the addition of oxaloacetate for reoxidation of the NADPH in the stroma. 6. Dark CO2 fixation in the presence of dihydroxyacetone phosphate and oxaloacetate has the same pH dependency as CO2 fixation in the light. This demonstrates that CO2 fixation in the dark is not possible, unless the pH in the medium is artificially raised to pH 8.8.

Adenosine Triphosphate↗
PubMed1976

Studies to improve fixation of human nerves. V. Effect of temperature, fixative and CaCl2 on density of microtubules and neurofilaments.

Utilizing morphometry of electron micrographs of nerves of rat fixed with different schedules of fixation it was concluded that: irrespective of the fixative, fixation at low temperature is associated with markedly low densities of microtubules (MT) of myelinated and of unmyelinated fibers; of the various fixatives tested 2% glutaraldehyde in 0.1 M cacodylate buffer without 0.025 M CaCl2 was associated with the highest density of MT of myelinated fibers and the addition of 0.025 M CaCl2 to a 2% glutaraldehyde fixative solution is associated with abnormally low densities of MT in myelinated and unmyelinated fibers. An effect of temperature, fixative and added CaCl2 on neurofilaments density was not demonstrated. Using the criterion of density of MT and assuming that these studies on rat nerves can be applied to biopsied nerve of man, fixation of human nerves should be at room or body temperature, and the fixative solution should not contain 0.025 M CaCl2.

Animals↗
PubMed1977

Comparison of immediate polyvinyl alcohol (PVA) fixation with delayed Schaudinn's fixation for the demonstration of protozoa in stool specimens.

Immediate fixation of stool specimens in polyvinyl alcohol fixative (PVA) was compared with Schaudinn's fixation delayed until the specimens were received in the laboratory, in a series of 100 consecutive positive stool specimens. More specimens were found positive following PVA fixation, and the numbers of organisms present on the slides were greater in specimens processed by this technique than after Schaudinn's fixation. It is concluded that immediate fixation results in the preservation of larger numbers of organisms in a recognizable state. The routine use of PVA fixation prior to transportation of the specimen to the laboratory is recommended.

Amebiasis↗
PubMed1975

Fixation of foetal pig liver for electron microscopy. I. The effect of various aldehydrs and of delayed fixation.

Light and electron microscopic investigations of the ability of various fixatives to preserve foetal pig liver tissue (immersion fixed 5 min, 20 min, and 40 min after the death of the mother) gave the following results: 1) Fixation with 4% formaldehyde gives, in all circumstances, an unacceptable tissue presevation. 2)Fixation with 2.5% glutaraldehyde gives fair tissue preservation in a ca. 130 mu thick zone below the surface. 3) Fixation with a fixative mixture containing 2% formaldehyde and 1.25% glutaraldehyde or 2% formaldehyde, 1% glutaraldehyde and 1% acrolein gives good tissue preservation in a ca. 500 mu thick zone under the surface. 4) The addition of DMSO does not improve tissue preservation. 5) Delayed fixation of liver tissue removed from the foetus after the uterus has been placed in an incubator for 40 min has only a slight effect on tissue preservation.

Acrolein↗
PubMedDate not supplied

[Kinetics of sodium thiosulfate-35S fixation and effect of some sulfur compounds on iodine-131 fixation in ray thyroid gland].

In order to invalidate or confirm the affirmation that non antithyroid sulphide molecules alter the measure of the thyroid fixation rate of iodine 131 we undertook on the rat: on one hand a kinetic study of thyroid fixation of sodium thiosulfate labelled with sulphur 35, which showed a very low captation not exceeding 0,01% of injected radioactivity; on the other hand the study of the effects of some sulphide molecules on thyroid fixation of iodine 131 in the rat: sodium thiosulfate, association of sodium thiosulfate + metalloidal sulphur + methionine, carbutamide and dimethylsulfoxyde in various kinds of dose administration and periods. None of the products used in our work conditions produced a significant decrease of the fixation rate of iodine 131. In three different experimental protocols (sulfur association and dimethylsulfoxide), we showed a significant light increase of the fixation rate.

Animals↗
PubMed1978

Microwave fixation versus formalin fixation of surgical and autopsy tissue.

Microwave irradiation of surgical and autopsy tissue penetrates, fixes, and hardens the tissue almost immediately (the fluid media used in the microwave consisted of saline, ten percent phosphate buffered formalin, and distilled water). Tissue sections from a representative sample of organs were tested. Comparable sections were simultaneously fixed in a phosphate buffered ten percent formalin bath in a vaccum oven as a control. Hematoxylin and eosin were used to stain the sections. Results equal to and superior to the control method were obtained. Saline microwave fixation was superior to formalin microwave fixation. Tissues placed in Zenker's solution and fixed in standard microwave oven (for approximately one minute) yielded results at least equal to conventional Zenker fixation (approximately two hours). No tissue hardening resulted from Zenker microwave fixation. A unique time versus temperature graph (microwave heating curve) reduces individual variation with this technique.

Autopsy↗
PubMed1979

Fixation of mammalian tissues in different fixatives and its influence on the staining with methyl green-pyronin.

This investigation is a study on the role of fixation of different mammalian tissues in different fixatives as well as in 10% neutral formalin containing different metal cations and its effect on the staining with the dye-mixture, methyl green-pyronin. The results indicate variation in the colour of the nuclei in tissues fixed in the different fixatives. The possible role of the fixatives on nuclear colouration has been discussed.

Animals↗
PubMed1976

[Comparison of the sensitivity of complement-fixation test and latex-fixation test in the demonstration of a migration phase of ascariasis (Ascaris suum) in a non-specific host].

The authors compared the sensitiveness of the complement-fixation reaction and the latex-fixation test in the demonstration of a migration phase of ascariasis in a non-specific host (rabbit). The sensitiveness of the latex-fixation test was higher by the value of one dilution of the geometric series, on an average.

Animals↗
PubMed1979

Differential fixation of poly(L-arginine) and poly(L-lysine) by tannic acid and its application to the fixation of collagen in electron microscopy.

A differential fixation of poly(L-arginine) and poly(L-lysine) has been demonstrated by means of cellulose acetate electrophoresis and colorimetric titration. Electrophoresis showed that at pH 3.0 and concentrations between 0.025% and 2% the reagent interacts with poly(L-arginine) but not with poly(L-lysine). At pH 7.5, however, poly(L-lysine) also reacts, although at a higher concentration of tannic acid than was required to fix poly(L-arginine) at this pH. Colorimetric titration revealed that for poly(L-arginine) the reaction with tannic acid commences at pH 3.0 and is complete at pH 4.1 whereas for poly(L-lysine) the reaction commences at pH 3.5 and is complete at pH 4.9. It is suggested that the reaction is predominantly electrostatic. The results are discussed in relation to the use of tannic acid as a protein fixative in electron microscopy.

Animals↗
PubMed1978

Fixatives and methods of fixation in selected tissues of the laboratory rat.

The histologic appearances of tissues fixed by immersion and perfusion were compared, as well as the effects of these different fixatives: 10% neutral buffered formalin, Carnoy's fluid, and Bouin's fluid. Intravascular perfusion provided better tissue preservation than fixation by immersion. The quality of preservation by neutral buffered formalin was equal to that of Carnoy's fluid and Bouin's fluid, but the overall staining quality of the latter two was judged to be superior.

Animals↗
PubMed1979

Mitotic activity and delay in fixation of tumour tissue. The influence of delay in fixation on mitotic activity of a human osteogenic sarcoma grown in athymic nude mice.

The purpose of the present investigation was to study the effect of delay in fixation on the mitotic activity in tumour tissue. A human osteogenic sarcoma, especially suitable for counting of mitoses, grown in athymic nude mice, was fixed with varying delay and the mitotic, prophase, metaphase and ana-telophase indices were determined. An almost exponential decline of the mitotic index was observed with a reduction to 49.4% and 15.0% after respectively 60 and 180 minutes. The proportional incidence of prophases, metaphases and ana-telophases changed so that a relative accummulation of advanced phases occured during the 180 minutes of observation. It is concluded that delay in fixation of a magnitude, which is not uncommon in routine surgical pathology, may allow the majority of mitoses to terminate, resulting in unreliable assessments of mitotic activity.

Adult↗
PubMed1976

Studies to improve fixation of human nerves. IV. Effect of time elapsed between death and glutaraldehyde fixation on density of microtubules and neurofilaments.

Glutaraldehyde fixation of rat peroneal nerves after the animal was killed (simulated autopsy) was associated with a statistically significant decrease of density (number per unit area) of microtubules (MT) in comparison with controls (simulated biopsy). The density of MT decreased progressively with prolongation of time elapsed prior to glutaraldehyde fixation (1, 6, and 12 hours). Such a time effect was not demonstrated for density of neurofilaments.

Animals↗
PubMed1979

[Fixation of the osteotomised greater trochanter by total hip prosthesis in 181 cases. Advantages of the fixation with intramedullary Kirschner wire and the figure-of-eight wire loop (author's transl)].

It is mostly possible to implant a total hip prosthesis without an osteotomy of the greater trochanter. It is however necessary to do it in some cases such as an ankylosed hip, a reimplantation and a severe form of the coxarthrosis. 181 patients have been followed up. The fixation by intramedullary Kirschner wire and figure-of-eight wire loop has been of success. It is a simple method. No fracture of the greater trochanter has been observed.

Aged↗
PubMed1978

Fixation of pelvic fractures and dislocations. An experimental study on the loading of pelvic fractures and sacro-iliac dislocations after external compression fixation.

A trapezoid external compression fixation frame, assembled with the Hoffman instruments, was used for stabilizing experimental injuries to the pelvic skeleton of ten cadaver specimens. The resistance to loading in a position corresponding to upright standing was tested in 17 experiments and related to the calculated load in vivo. The results indicated that ipsilateral injuries, either presenting as dislocations of the sacro-iliac joint and symphysis or as unilateral fractures of the sacrum or ilium in combination with fractures of the pubic rami could be stabilized by the external compression frame well enough to permit weight-bearing in the upright standing position. Bilateral injuries to the pelvic skeleton, vertical or oblique, could not, however, be stabilized enough to resist more than a fraction of the normal load in the upright standing position.

Aged↗
PubMed1978

The effect of limited fragmentation of porcine 19 S IgM on agglutination and complement fixation. Evidence that at least two intact subunits are required for complement fixation by IgM antibody.antigen complexes.

Limited fragmentation of pig IgM anti-Salmonella with pepsin gave a 12 S product which agglutinated Salmonella almost as efficiently as the original IgM but the complement fixing ability of the complexes was substantially lowered. The 12 S product was shown to consist of molecular entities having intact Fc regions with an average of 3 or 4 intact Fab arms still attached. Our results appeared to show that removal of Fab arms by pepsin was a random process and on this basis a mathematical model was set up. From this model it was possible to calculate the frequencies of molecular entities which would give rise to the observed average of 3 or 4 Fab arms. The most likely explanation of our results is that complexes need to be formed by molecules having at least two intact subunits before efficient complement fixation takes place.

Animals↗