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At least 217 records · Page 12Linked to original sources
PubMed1988

Immunological typing of acute leukemias: immunoenzymatic staining of fixed cells compared with immunofluorescence staining of unfixed cells in suspension.

A panel of 14 monoclonal antibodies (McAb) against hematopoietic cell surface antigens was applied on mononuclear blood or bone marrow cells from 40 cases of acute leukemia in order to compare immunoenzymatic staining (IE) (alkaline phosphatase) of fixed cells with immunofluorescence staining (IF) of unfixed suspended cells. According to the immunological results, 25 cases were phenotyped as ALL and 15 cases as AML. Cases with blast crisis secondary to chronic myelogenous leukemia (CML-BC) were not represented in this series. In all ALL cases the two methods gave an identical antigenic distribution. In 20 our of 21 cases of non-T-cell ALL, a B-cell progenitor origin was demonstrated by a positive staining reaction with the anti-CD19 McAb AB1 or HD37, and in 10 cases additionally with the anti-CD20 McAb B1 or 1F5. In contrast to the results obtained with IF, IE revealed a poor preservation of the AB1 epitope on CD19, whereas the HD37 epitope was equally well demonstrated by both methods. In 15 cases of AML the distribution of positive versus negative cells with IE or IF was identical for all McAb except J5 (anti-CALLA) (CD10) and B1 (CD20). Thus, 10/15 AML cases expressed CALLA with IE compared to 2/15 with IF. The corresponding figures for B1 were 5/15 and 0/15, respectively. Accordingly, normal myeloid precursor cells were CALLA-positive with IE but negative with IF. The discrepancy probably reflects the fact that, whereas both intracytoplasmatic and membrane-bound antigens are exposed in IE, only the latter are in IF. If the alteration of antigenic accessibility after fixation is considered, IE can safely be used for immunophenotyping of acute leukemia.

Antibodies, Monoclonal↗
PubMed1983

Chemical mechanism of the Gram stain and synthesis of a new electron-opaque marker for electron microscopy which replaces the iodine mordant of the stain.

Crystal violet (hexamethyl-para-rosaniline chloride) interacts with aqueous KI-I2 during the Gram stain via a simple metathetical anion exchange to produce a chemical precipitate. There is an apparent 1:1 stoichiometry between anion (I-) and cation (hexamethyl-para-rosaniline+) during the reaction and, since the small chloride anion is replaced by the bulkier iodide, the complex formed becomes insoluble in water. It is this same precipitate which forms in the cellular substance of bacteria (both gram-positive and gram-negative types) and which initiates the Gram reaction. Potassium trichloro(eta 2-ethylene)-platinum(II), as an electronopaque marker for electron microscopy, was chemically synthesized, and it produced an anion in aqueous solution which was compatible with crystal violet for the Gram stain. It interacted with crystal violet in a similar manner as iodide to produce an insoluble complex which was chemically and physically analogous to the dye-iodide precipitate. This platinum anion therefore allows the Gram staining mechanism to be followed by electron microscopy.

Bacillus subtilis↗
PubMed1979

The cationic carbocyanine dyes Stains-all DBTC, and Ethyl-Stains-all, DBTC-3,3',9 triethyl.

A simple method is presented for distinguishing two closely related metachromatic carbocyanine dyes: Ethyl-Stains-all, a triethyl dye, and Stains-all, a diethyl methyl dye. This has become important since one lot of the triethyl dye was distributed erroneously under the diethyl methyl label. The dyes differ in solubility and in differential staining of macromolecules. Studies performed with both dyes are summarized.

Carbocyanines↗
PubMed1994

Double epi-illumination microscopy with separate visualization of two antigens: a combination of epi-polarization for immunogold-silver staining and epi-fluorescence for alkaline phosphatase staining.

We present a method for an epi-illumination immunohistochemical double staining approach. The method combines the use of an immuno-alkaline phosphatase technique and the immunogold-silver technique, visualized with epifluorescence and epi-polarization illumination, respectively. Out of six tested alkaline phosphatase activity-revealing methods, only the reaction product obtained with the Becton Dickinson CAS Red kit showed an intense red fluorescence with a rhodamine filter set and no signal with epi-polarization illumination. The silver precipitate did not exhibit any signal with the rhodamine filter set. This allows separate observation and photographic recording of two antigens in one tissue section, an objective that cannot be achieved with conventional immunoenzyme double staining methods. The double epi-illumination approach presented is compatible with different immunoenzyme double staining protocols.

Alkaline Phosphatase↗
PubMed2001

Staining pattern of the brush border and detection of cytoplasmic granules in the uriniferous tubules of female DBA/2Cr mouse kidney: comparison among various fixations and stains.

The proximal straight tubules of the female mouse kidney exhibit heavy periodic acid Schiff (PAS) staining in their brush borders and numerous cytoplasmic granules. In the present study, the female DBA/2Cr mouse kidney was examined, using various fixatives (formalin, PFA, PLP, Zamboni's, Bouin, or Carnoy solution) and various staining methods (HE, PAS, alcian blue, periodic acid methenamine-silver (PAM), toluidine blue, azan, or Congo red). Under azan and PAM, the staining pattern of the brush border was similar to that of PAS, and few effects of the fixative were observed. Cytoplasmic granules were clearly detected with PAM as well as PAS. However, these granules were not detected with Carnoy solution. Furthermore, distribution of granules differed between PAS and PAM.

Animals↗
PubMed1988

Staining of the nucleolus with protein and RNA stains for automatic measurement of nucleolar size in paraffin sections.

In this paper two staining methods for automatic assessment of nucleolar profile area in paraffin sections of breast carcinomas using the IBAS 2000 image analyzer are reported. In the first method, the nucleolar proteins are stained with phloxine B, in the second the nucleolar RNA is stained with methyl green-pyronin. Both methods are fully reproducible. Statistically, the profile areas observed for the same patient by the two methods were found not to differ significantly. Because of their rigidity and their size in relation to section thickness, nucleoli are particularly attractive cell structures for quantitation, for instance in studies of variables that might be useful in establishing the prognosis for carcinoma patients.

Breast Neoplasms↗
PubMed2002

[The relationship of cholesterol (CHO) and triglycerides (TG) in VLDL, LDL, and VLDL subfractions by agarose gel electrophoresis, using CHO and TG dual staining (AG-CHO, TG staining)].

The measurement of CHO and TG concentrations in three lipoprotein subfractions (VLDL, LDL, and HDL) are useful to estimate qualitative change of lipoproteins. This method, CHO and TG dual staining using agarose gel electrophoresis (AG-CHO, TG staining), is conventional and the results correlated highly with ultracentrifugation. Using this method, we measured CHO and TG concentrations of VLDL, LDL, and HDL subfractions in 244 patients after an overnight fast. All cases were stratified to 4 groups, normolipidemia (LDL-CHO < 160 mg/dl, sTG < 150 mg/dl, n = 111), type IIa(n = 34), IIb(n = 39), and IV(n = 36), according to hyperlipidemia types. Furthermore, normolipidemia with low HDL cases (HDL-CHO < 45 mg/dl, n = 24) distinguished from HDL-CHO > or = 45 mg/dl, normolipidemia cases(n = 111). Between serum TCHO and LDL-CHO, serum TG and VLDL-CHO, TG showed positive correlation(n = 0.895, 0.971), particularly serum TG and VLDL-TG showed strong positive in all lipidemia types. However, serum TG and VLDL-CHO was strongly positive in type IIb and IV(r = 0.825, 0.823), but weakly positive in type IIa(r = 0.459). HDL-CHO showed no correlation with sTCHO and sTG. The correlation of CHO and TG with each subfraction was strongly positive in VLDL(r = 0.910), weakly positive(r = 0.49) in LDL, and showed the no correlation in HDL in all cases. These correlation varied in lipidemia types, IIb and IV were strongly positive(r = 0.886, 0.838) in VLDL subfraction, but nomolipidemia cases(r = 0.555) showed significant weaker correlation(p < 0.0001). In the LDL subfraction, IIb and IV showed no correlation(r = 0.009, 0.163) between CHO and TG. The CHO/TG ratio of three subfractions were widely distributed, and type IIb and IV distributed to lower range than normolipidemia and/or type IIa lipidemia in three subfractions. From these results, dual measurement of LDL-CHO and LDL-TG are useful to estimate qualitative change in the LDL subfraction. For instance, in high LDL-CHO or LDL-TG with low CHO/TG cases, we could suspect the presence of IDL-rich particles in the LDL subfraction or small particle LDL cases. HDL-CHO and CHO/TG show positive(r = 0.714), HDL-CHO and VLDL-TG, VLDL-CHO showed weakly negative correlations(r = 0.500, 0.487), showing that high HDL-CHO level cases tended to have a CHO rich, and low VLDL-TG concentration. These results indicated that qualitative change in lipoproteins could be clarified by measurement of the TG concentration, in addition to CHO concentrations in three subfractions. We conclude that AG-CHO, TG staining method is useful for diagnosis and monitoring of dyslipidemia.

Adult↗
PubMed1987

Standard specimens for stain calibration and their application to the Papanicolaou stain.

Standardized specimens composed of extracts of biologic objects (nucleoprotamine and bovine liver) were developed as tools for the quantitative evaluation of stain performance on biologic substrates. The specimens are mixtures of proteins and nucleic acids and thus mimic the staining characteristics of cytologic smears. The concentration of each mixture and the specimen thickness can be precisely controlled, ensuring the production of a large number of samples with a nearly identical capability for dye binding. The transmitted light spectra of the standardized specimens varied depending on the extract and the preparation conditions. Spectra similar to those reported from the nuclei and cytoplasm of cell types in Papanicolaou-stained cervicovaginal smears were observed. Light transmission was uniform to +/- 5% across each specimen and from specimen to specimen. The specimen thickness was uniform within +/- 2%. Studies with these standardized samples could reveal the much-needed correlations between the chemical and optical characteristics of dyes and dye solutions and the performance of the dyes on biologic substrates.

Animals↗
PubMed1968

Quantitative studies of immunofluorescent staining. Relationships of characteristics of unabsorbed antihuman IgG conjugates to their specific and non-specific staining properties in an indirect test for antinuclear factors.

Reproducible titres of indirect immunofluorescent (IF) staining with antinuclear factor (ANF)-containing sera could be obtained with different antihuman IgG conjugates by quantitative adjustments of their characteristics. Conversely, one ANF yielded a broad range of ANF titre (80-640) upon appropriate adjustments of the conjugate characteristics. The same and related characteristics of the conjugates also afforded a basis for quantitatively defining the conditions under which non-specific staining (NSS) appeared. The salient characteristics of the anti-IgG conjugates include: (1) their strength of antiglobulin (expressed as units/ml of precipitating antibody or mug antibody N/ml); (2) their apparent fluorescein concentration (in mug F/ml); (3) their protein concentration (in mg/ml). Optical and immunologic sensitivity ratios are calculated from these conjugate characteristics. Optical sensitivity (expressed as fluorescein concentration to protein concentration (F/P) ratios), immunological sensitivities (expressed as units/1% protein) and the dilution employed serve to characterize quantitatively anti-IgG conjugates adequately to define their specific and non-specific staining properties.

Antibodies, Antinuclear↗
PubMed1980

Quantitative studies on immunofluorescent staining. VII. Fractionation of conjugates by acid dialysis to reduce nonspecific staining.

Dialysis of fluorescein-conjugated antibodies at pH 5.8-6.0 can reduce unwanted nonspecific staining. Selective precipitation of highly fluoresceinated proteins in mildly acidic buffer is a simple technique which can produce a reagent with greatly improved nonspecific staining characteristics with little or no loss of antibody activity. This technique may be especially useful when fluorescent antibody tests are to be performed on tissues which have a high propensity for nonspecific staining such as skin.

Dialysis↗
PubMed1980

Backscatter electron imaging (BEI) of paraffin sections stained with heavy metal histopathologic stains, with observations on some variables encountered in BEI.

Using a recently developed annular photolithographic type backscatter detector, we studied a number of heavy metal stains used routinely in light microscopy. These were paraffin sections fixed with standard light microscopic fixation and dehydration techniques with no special modification made to them prior to coating of the slides with carbon for scanning observations. It was found that many of the stains that work in light microscopy will also provide good results in backscatter imaging if the differentiation of the staining is good. Studying the effects of varying accelerating voltages, it was discovered that optimum results and the most reliable data are achieved by comparative examination of the specimen at both a low and high voltage. This is necessary to prevent misinterpretation of data as a result of inadequate beam penetration of the sample.

Animals↗
PubMed1977

Repair in arterial tissue. The role of endothelium in the permeability of a healing intimal surface. Vital staining with Evans blue and silver-staining of the aortic intima after a single dilatation trauma.

The role of the endothelium as a factor determining the permeability of a healing aortic intima after a severe mechanical lesion in the rabbit was studied by vital staining with Evans blue, by surface light microscopy after silver-staining, fluorescence microscopy and by other methods. Endothelium to develop from pre-existing endothelium in the mouths of intercostal arteries and at the borders of the lesion, remained unstained by vital staining, whereas denuded surfaces as well as pseudo-endothelium, probably derived from smooth muscle cells in the neointima, were intensely coloured. There was a general tendency towards a decreasing permeability of the aortic wall throughout an experimental period of 6 months. The determinant factor in the decreasing permeability was the presence of genuine endothelium, but changes in the endothelium, probably in the form of differentiation of intercellular junctions and changes in the connective tissue in the deeper layers, seemed to be additional factors which influenced the permeability.

Animals↗
PubMed1993

Evaluation of sensitivity and specificity of cold stain vis-a-vis Ziehl Neelsen stain on culture results.

Various cold stain techniques have been tried with varying success since early part of the century. Dr. S. Kudoh of Japan and Dr. A. J. Pathan of Pakistan discussed to evolve a simple process for cold method (see the foot note) which is a simple procedure for staining tubercle bacilli in sputum specimen. And the cold method was found as sensitive, as specific and almost as reliable as Ziehl Neelsen on direct comparison of two techniques. In this paper, the cold method was further evaluated on culture results with Ziehl Neelsen stain during 1987-90 in our institute.

Cold Temperature↗
PubMedDate not supplied

The alcohol-fixed Diff-Quik stain. A novel rapid stain for the immediate interpretation of fine needle aspiration specimens.

A rapid alcohol-fixed Diff-Quik stain is described. The technique takes less than five minutes and results in excellent cytologic detail, comparable to that with Papanicolaou stain, as well as superb stromal definition. The stain should be very useful to pathologists and cytotechnologists for immediate interpretation of fine needle aspiration specimens.

Azure Stains↗
PubMed1996

Staining system: a cheap and simple way for staining ultrathin sections.

A staining device for sectioned material supported on grids for viewing at the Transmission Electron Microscope is described. The method based in a double side sticky tape is inexpensive, rapid and clean. By using only pipettes and a double side sticky tape the best solution to the tedious problems of sections staining is obtained. The tape is discarded and the staining solution too. Precipitates have not been observed.

Microscopy, Electron↗
PubMed1984

[Hydrophobic acridine dyes for fluorescence staining of mitochondria in living cells. 2. Comparison of staining of living and fixed Hela-cells with NAO and DPPAO].

The hydrophobic fluorescence dyes NAO and DPPAO (see scheme of structural formulae) stain the mitochondria of living HeLa-cells. The trans-membrane potential favours the dye accumulation of the cation NAO and supports the hydrophobic interaction of the dye with the mitochondrial membrane lipids and proteins. The lecithin-like dye DPPAO is electrical neutral. Its binding to mitochondria of living cells is only caused by hydrophobic interaction. NAO and DPPAO stain also the mitochondria of glutaraldehyde fixed HeLa-cells in aqueous medium. Fluorescence staining occurs even after extraction of the lipids of the cell with acetone. We suppose that the dye accumulation in the mitochondria of the fixed cells is caused by the hydrophobic interaction between the dyes and the very hydrophobic mitochondrial lipids and proteins.

1,2-Dipalmitoylphosphatidylcholine↗
PubMed1981

Fluorescent staining of neuromuscular junctions by using the antibody against acetylcholine receptors of Narke japonica, and double staining with the antibody and erabutoxin b.

The neuromuscular junctions of various vertebrates were visualized by indirect immunofluorescence microscopy using antibody against purified acetylcholine receptors (AChRs) of the electric organ from Narke japonica. Further, by using rhodamine-labeled erabutoxin b (TMR-Eb), we showed that AChRs at the neuromuscular junctions of frog, chick and mouse muscle could not be doubly stained with the antibody and erabutoxin b. AChRs of snake muscle could not be stained with TMR-Eb, while they were stained with the antibody against AChR. Moreover, the antibody did not inhibit the binding of 3H-labeled erabutoxin b to the solubilized AChRs from the mouse muscle. These results indicate that, as far as the antibodies against Narke AChRs are concerned, most antibody-binding sites in the molecules of muscle AChR in situ are different from those responsible for binding of the snake neurotoxin.

Acetylcholine↗