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PubMed1993

Selective loss of human leukocyte class I allospecificities and staining of melanoma cells by monoclonal antibodies recognizing monomorphic determinants of class I human leukocyte antigens.

Immunoperoxidase staining of frozen sections from surgically removed melanoma lesions showed that anti-human leukocyte antigen (HLA)-A2, A28 monoclonal antibody (mAb) stained keratinocytes, but did not stain melanoma cells in 21% of the 14 primary and 44% of the 9 metastatic lesions tested. The loss of HLA-A2 and/or A28 allospecificities did not affect the staining patterns with mAb recognizing monomorphic determinants of HLA Class I antigens, in terms of percentage of stained melanoma cells and intensity of staining. This finding is not likely to reflect the sensitivity of the immunoperoxidase technique, since cytofluorographic analysis detected no significant difference in the staining pattern by mAb to monomorphic determinants of HLA Class I antigens between a melanoma cell line and an autologous transfectant that had acquired HLA-A2 antigens following gene transfer. The results of the present study imply that the frequency of abnormalities in HLA Class I antigen expression by melanoma cells is higher than that described in the literature, since selective losses of HLA Class I allospecificities are not detected by staining of melanoma cells with mAb to monomorphic determinants of HLA Class I antigens. The latter reagents have been used in most of the published studies to characterize the expression of HLA Class I antigens in melanoma lesions. Furthermore, the present results provide a mechanism for the unexpected resistance to cytotoxic T-cell-mediated lysis and the unexpected poor clinical course of the disease in some patients despite a high expression of HLA Class I antigens as measured by staining of melanoma cells with mAb to monomorphic determinants.

Antibodies, Monoclonal↗

PubMed1995

The effect of an oscillating/rotating electric toothbrush and a sonic toothbrush on removal of stain from enamel surfaces.

This laboratory investigation compared the Sonicare toothbrush, the Braun Oral-B Plaque Remover and a manual brush, in terms of removal of stained pellicle from the surface of enamel specimens. Enamel specimens were stained over a period of 4 days to produce a stained pellicle film. Staining was assessed photometrically before and after brushing 16 specimens per group for 4.5 minutes with the three test toothbrushes and dentifrice slurry prepared from Crest toothpaste. The positive control consisted of the ADA reference abrasive and a manual toothbrush. Results were expressed as a cleaning ratio (CR), with higher values representing more efficient stain removal. Analysis of results revealed that removal of stain from the enamel surfaces was similar in the Braun Oral-B Plaque Remover group (CR 94) and the reference standard group (CR 100). There was no statistically significant difference between these 2 groups. In contrast, stain removal in the Sonicare toothbrush group and the manual toothbrush group was significantly less effective (p<0.01), with a mean CR of 66 and 76, respectively. There was no statistical difference between the cleaning ratio scores for these 2 groups. It is concluded that extrinsic stain removal with the Sonicare toothbrush is similar to that achieved with a manual brush, but that stain removal with the Braun Oral-B Plaque Remover is significantly more effective.

Analysis of Variance↗

PubMed1995

Laboratory and clinical stain removal evaluations of two tartar control dentifrices.

The purpose of this research was to evaluate Colgate original Tartar Control dentifrice (TC) and new Colgate Micro Cleansing Tartar Control (MCTC) dentifrice in a laboratory and clinical study for their ability to remove induced stain. In the laboratory study, polished and etched bovine enamel specimens were stained for 4 days with a coffee, tea, and mucin, Sarcina lutea tartox culture in trypticase soy broth. Sixteen specimens were then brushed for 200 strokes with a sodium carboxymethyl cellulose solution to remove loose stain, and then brushed for 300 strokes with a 1:1 toothpaste slurry. Tooth color was measured by reflectance with the CLE L*a*b* scale. The calculated stain removal, 37% for MCTC and 24% for TC, was significantly different (p<0.01) favoring the MCTC dentifrice. In the clinical study, eight-six subjects were given Peridex (0.12% chlorhexidine gluconate) mouthrinse, a low abrasive dentifrice and a soft toothbrush to use twice a day for 8 weeks. Stain was monitored on the buccal and lingual surfaces of the anterior teeth using the Lobene index. The seventy-seven subjects with sufficient total stain were stratified into two balanced groups, given a new soft toothbrush and were randomly assigned to use one of the dentifrices tested, twice a day for 8 weeks. At 4, 6 and 8 weeks in the study, the MCTC dentifrice product had consistent, significantly (p<0.05-0.001) lower total stain scores than the TC dentifrice. In the laboratory and clinical studies conducted, even though stain induction and evaluation procedures differed, the new Colgate Micro Cleansing Tartar Control dentifrice was significantly more effective at removing stain than the Colgate original Tartar Control dentifrice formula.

Adolescent↗

PubMed1987

Understanding Romanowsky staining. I: The Romanowsky-Giemsa effect in blood smears.

Normal blood smears were stained by the standardised azure B-eosin Y Romanowsky procedure recently introduced by the ICSH, and the classical picture resulted. The effects of varying the times and temperature of staining, the composition of the solvent (buffer concentration, methanol content, & pH), the concentration of the dyes, and the mode of fixation were studied. The results are best understood in terms of the following staining mechanism. Initial colouration involves simple acid and basic dyeing. Eosin yields red erythrocytes and eosinophil granules. Azure B very rapidly gives rise to blue stained chromatin, neutrophil specific granules, platelets and ribosome-rich cytoplasms; also to violet basophil granules. Subsequently the azure B in certain structures combines with eosin to give purple azure B-eosin complexes, leaving other structures with their initial colours. The selectivity of complex formation is controlled by rate of entry of eosin into azure B stained structures. Only faster staining structures (i.e. chromatin, neutrophil specific granules, and platelets) permit formation of the purple complex in the standard method. This staining mechanism illuminates scientific problems (e.g. the nature of 'toxic' granules) and assists technical trouble-shooting (e.g. why nuclei sometimes stain blue, not purple).

Azure Stains↗

PubMedDate not supplied

Viability and acrosome staining of stallion spermatozoa by Chicago sky blue and Giemsa.

A simple trypan blue-neutral red-Giemsa staining procedure for simultaneous evaluation of acrosome, sperm head, and tail membrane integrity and morphology has been used to evaluate equine spermatozoa. Some special characteristics and problems have arisen in evaluating stallion semen. One problem was the differentiation of intact vs. damaged sperm tails primarily in frozen and thawed samples. After freezing and thawing, a high percentage of spermatozoa with an unstained head and stained tail were observed. These cells are considered immotile. Therefore, unambiguous differentiation of intact vs. damaged sperm tail membrane is very important for evaluating semen quality. The aim of our study was to develop a method especially for stallion sperm to distinguish more accurately the different cell types. We compared Chicago sky blue 6B (CSB) to trypan blue (TB) for viability staining. CSB/Giemsa staining showed good repeatability and agreement with TB/Giemsa measurements. For densitometry analysis, individual digital images were taken from smears stained by CSB/Giemsa and by TB/Giemsa. A red-green-blue (RGB) histogram for each area of spermatozoa was drawn. Differences of means of RGB values of live vs. dead tails and separate live vs. dead heads from each photo were used to compare the two staining procedures. CSB produced similar live/dead sperm head differentiation and better tail differentiation. TB can be replaced by CSB and this results in more reliable evaluation. After staining with 0.16% CSB and 4 min fixation, 2-4 h Giemsa staining at 25-40 degrees C is recommended for stallion semen.

Acrosome↗

PubMed1980

A microdensitometric method for the analysis of staining kinetics.

The rate of histological staining is in general controlled by diffusion of dye in the dyebath and/or substrate. Microdensitometric and mathematical methods for studying the rate of staining are described, and illustrated by experiments on the staining of pancreatic cytoplasmic RNA and nuclei in sections by Azure at pH 4. Under given conditions the half-staining time, t1/2, of RNA was about twice that of nuclei, reflecting differences in permeability or porosity. Half-staining times were approximately doubled if the concentration of dye in the bath was reduced by 75%, suggesting that staining is not a simple first-order process; one possible explanation for this is that rate is affected by dye diffusion both in the dyebath and in the substrate. Lowering the temperature from 298 K to 277 K increased t1/2 for RNA about eightfold; the activation energy E of staining was therefore approximately 68 kJ mol-1. Assuming the substrate to consist either of plane sheets of thickness b, or uniform cylinders of radius r, equations originally proposed by Crank were applied to rate-of-dyeing data to calculate D/b2 and D/r2, where D is the diffusion coefficient of dye inside the substrate. From the observation that D/b2 and decreased markedly during the staining process it was concluded that both substrates studied were heterogeneous.

Animals↗

PubMed1975

An evaluation of some commerical Romanowsky stains.

The staining properties of 43 commerical Romanowsky-type stains have been studied. Considerable differences in the appearance of stained blood films were observed with different batches of these stains, the staining of red cells being particularly variable. Attempts have been made to correlate staining patterns with stain composition as revealed by thin-layer chromatography and sulphated ash analyses. In this way it has been possible to define some essential requirements for satisfactory staining.

Azure Stains↗

PubMed2006

Effect of staining procedures on the results of micronucleus assays with exfoliated oral mucosa cells.

Micronuclei in exfoliated epithelial cells are widely used as biomarkers of cancer risk in humans. To elucidate the effect of different staining procedures on the outcome of such investigation, we conducted a study in which the micronuclei frequencies in oral mucosa cells of heavy smokers (n = 20) and nonsmokers (n = 10) were evaluated with nonspecific (Giemsa, May-Grünwald-Giemsa) and DNA-specific (4',6-diamidino-2-phenylindole, Feulgen, acridine orange) stains, whereas with Giemsa-based stains, the frequencies of micronuclei in smokers were significantly (4- to 5-fold) higher in the smokers group, no significant increase was observed with any of the DNA-specific stains. Furthermore, the evaluation of cells of the two study groups with Feulgen stain showed that oral mucosa cells from smokers had significantly increased levels of nuclear anomalies other than micronuclei. These anomalies are consequences of cell injury found in epithelial cells and are paralleled by formation of keratin bodies in the cytoplasm that resemble micronuclei. Correlation analyses showed that micronuclei frequencies scored in Giemsa-stained slides correlated significantly with karyorrhexis, karyolysis, condensed chromatin, and binucleates, whereas no such correlations were found with DNA-specific stains. These findings indicate that nuclear anomalies (and possibly keratin bodies) may be misinterpreted as micronuclei with nonspecific DNA stains and lead to false-positive results in studies with cells of epithelial origin. Furthermore, our results show that exposure of oral mucosa cells to genotoxic carcinogens contained in tobacco smoke does not lead to induction of micronuclei in these cells.

Adult↗

PubMedDate not supplied

Application of Ultrafast Papanicolaou stain to body fluid cytology.

OBJECTIVE: To investigate the applicability of the Ultrafast Papanicolaou stain to the cytology of fluids and to compare it with other methods. STUDY DESIGN: Over a 30-month period, 528 unfixed fluids (462 serous effusions, 48 pelvic washings, 16 cyst fluids and 2 bile duct drain fluids) were mixed thoroughly and centrifuged. Two Swedish-style air-dried smears were made and stained with Diff-Quik (Mercedes Medical, Inc., Sarasota, Florida, U.S.A.) and Ultrafast Papanicolaou stain (Richard Allan Scientific, Kalamazoo, Michigan, U.S.A.), and the remaining sediment was fixed in CytoRich Red (TriPath Imaging, Inc., Burlington, North Carolina, U.S.A.), centrifuged onto a 17.5-mm circle with a Hettich cytocentrifuge and stained by the Papanicolaou method. RESULTS: For the 115 malignant fluids, Ultrafast Papanicolaou stain was the preferred method in the 94 non-hematopoietic malignant fluids, Diff-Quik was the preferred method in the 9 hematopoietic malignancies, and CytoRich Red was the preferred preparation in 8 bloody effusions containing rare cancer cells and 4 malignant pelvic washings. The diagnostic turnaround time of smears stained by Ultrafast Papanicolaou stain was < 15 minutes, fast enough for intraoperative consultations. CONCLUSION: It seems that Ultrafast Papanicolaou stain improves the resolution of cytoplasmic and nuclear details of nonhematopoietic cells in body fluids. However, to detect cancer in all types of fluids, Diff-Quik and CytoRich preparations are also required. We now examine three slides per fluid sample, one slide by each of the three techniques.

Ascitic Fluid↗

PubMed1998

Histologic assessment of Helicobacter pylori status after therapy: comparison of Giemsa, Diff-Quik, and Genta stains.

To compare the Giemsa, Diff-Quik, and Genta stains for detection of Helicobacter pylori in gastric biopsy specimens, we stained, coded, and randomized slides, which were then independently reviewed by two pathologists and one trainee. H. pylori was graded from 0 (absent) to 5 (maximal intensity). Negative cases were from recently cured patients to ensure that a background of chronic inflammation was present. The time required to complete the evaluation was tabulated. The pathologists interpreted 72 H. pylori-negative slides, of which 1 (1.3%), 2 (3%), and 3 (4%) were scored positive (each with the score of 1) with Diff-Quik, Genta, and Giemsa stains, respectively (P = ns). Of the 128 H. pylori-positive slides, 5 (4%) had false-negative results with Diff-Quik, 8 with Genta (6%), and 14 with Giemsa stains( 11%). No Grade 2 slides were missed by Genta or Diff-Quik stains, but 3 of 20 were missed by Giemsa stain. The combination of hematoxylin and eosin and Diff-Quik provides accuracy similar to that of the Genta stain but requires more processing time. No stain was excellent after therapy for detecting one or two bacteria per slide; this finding emphasizes the need for obtaining multiple biopsy specimen to exclude failure of H. pylori therapy.

Azure Stains↗

PubMed1998

Development of calcium-permeable alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors in cultured neocortical neurons visualized by cobalt staining.

The developmental expression of calcium (Ca2+)-permeable alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) and kainate receptors in cultured neocortical neurons was evaluated by using cobalt uptake, a histochemical method that identifies cells expressing Ca2+-permeable, non-N-methyl-D-aspartate (non-NMDA) receptors. At a concentration of 500 microM, AMPA was found to stimulate cobalt uptake only late in development, resulting in staining of 2.7%+/-0.3% of the neurons maintained in culture for 12 days in vitro (DIV). When AMPA receptor desensitization was blocked with 50 microM cyclothiazide, the developmental profile of cobalt uptake mediated by 25 microM AMPA changed dramatically. The cobalt staining now appeared in young cultures (5 DIV), and the percentage of stained cells increased from 3.4%+/-0.2% at 5 DIV to 21.7%+/-1.6% at 12 DIV. The effect of 200 microM kainate was similar to that seen with 25 microM AMPA plus 50 microM cyclothiazide, resulting in 17.7%+/-0.3% stained neurons at 12 DIV. The cobalt uptake was specific to AMPA and kainate receptors because NMDA receptors and voltage-gated calcium channels were found not to mediate any cobalt staining. In addition, 10 microM 6-nitro-7-sulphamoylbenzo-[f]-quinoxaline-2,3-dione (NBQX) was able to prevent all staining at 5 and 8 DIV and most of the staining at 12 DIV, indicating that the non-NMDA ionotropic glutamate receptors are involved in cobalt uptake into the neurons. The AMPA receptor-selective antagonist GYKI 53655 was used to differentiate between cobalt influx through AMPA- or kainate-preferring receptors. After pretreatment with concanavalin A (con A), an inhibitor of kainate receptor desensitization, cobalt uptake was assessed after stimulation by 200 microM kainate in the presence of 25 microM GYKI 53655. No cobalt staining was observed under these conditions, indicating that most if not all of the cobalt influx induced by kainate was mediated through AMPA receptor channels.

Animals↗

PubMed1998

Gram stain method shows better sensitivity than clinical criteria for detection of bacterial vaginosis in surveillance of pregnant, low-income women in a clinical setting.

OBJECTIVE: The purpose of the study is to determine whether the Gram stain method is superior to the clinical criteria for the diagnosis of bacterial vaginosis in low-income pregnant women seen in a resident clinic setting. The clinical criteria is the current diagnostic method employed to diagnose bacterial vaginosis. STUDY DESIGN: In this study, 51 pregnant women with vaginal discharge were prospectively evaluated. All were screened using the clinical criteria, Gram stain method, and culture of the discharge. The modified scoring system instituted by Nugent et al. (J Clin Microbiol 29:297-301, 1991) was employed in reading the Gram stain smears. The clinical criteria were then compared with the Gram stain method. Isolation of moderate to many Gardnerella vaginalis growth by culture was used as the confirmatory finding. RESULTS: Sensitivity of the Gram stain method (91%) was significantly higher than that of the clinical criteria (46%), (sign test P = 0.0023, < 0.01). The Gram stain method also has both a low false-negative (4%) and high negative predictive value (96%), making it an ideal diagnostic test. CONCLUSION: The Gram stain method is a rapid and cost-effective test that is also highly reproducible and readily available in many laboratories. These features make the Gram stain method a more desirable screening procedure for bacterial vaginosis in a clinic population.

Cost-Benefit Analysis↗

PubMed2001

An improved, luminescent europium-based stain for detection of electroblotted proteins on nitrocellulose or polyvinylidene difluoride membranes.

SYPRO Rose Plus protein blot stain is an improved europium-based metal chelate stain for the detection of proteins on nitrocellulose and poly(vinylidene difluoride) (PVDF) membranes. Staining is achieved without covalently modifying the proteins. The stain may be excited with a 254 nm (UV-C), 302 nm (UV-B), or 365 nm (UV-A) light source and displays a sharp emission maximum at 612 nm. The emission peak has a full width at half-maximum of only 8 nm. The stain exhibits exceptional photostability, allowing long exposure times for maximum sensitivity. Since the dye is composed of a europium complex, it has a long emission lifetime, potentially allowing time-resolved detection, greatly reducing background fluorescence. Proteins immobilized to a nitrocellulose or PVDF membrane by electroblotting, dot-blotting, or vacuum slot-blotting are incubated with SYPRO Rose Plus protein blot stain for 15-30 min. Membranes are rinsed briefly, visualized with UV epi-illumination and the luminescence of the europium dye is measured using a 490 nm long-pass or 625 +/- 15 nm band-pass filter in combination with a conventional photographic or charge-coupled device (CCD) camera system. Alternatively, the dye may be visualized using a xenon-arc illumination source. The stain is readily removed from proteins by incubating membranes at mildly alkaline pH. The reversibility of the protein staining procedure allows for subsequent biochemical analyses, such as immunoblotting and biotin-streptavidin detection using colorimetric, direct fluorescence or fluorogenic visualization methods.

Blotting, Western↗

PubMed1978

PAS-lead hematoxylin as a stain for small-granule endocrine cell populations in the lungs, other pharyngeal derivatives and the gut.

Epithelial cells of the several subtypes that comprise the small-granule cell population of the respiratory system are little studied, partly because adequate silver, monoamine fluorescence and other specific light microscopical preparations have been more difficult to obtain than in the gut and other organs possessing diffuse endocrine systems. Periodic acid-Schiff (PAS) in combination with MacConaill-Solcia's lead hematoxylin has in our hands proven dependable for routine staining of serial 2-micrometer glycol methacrylate sections used in mapping the distributions of these cells along the airway. In lungs of mice, hamsters, kittens, and fetal rabbits, typical small-granule cells stain weakly or not at all with lead hematoxylin alone, hence are easily overlooked. PAS adds to the cytoplasm a diffuse magenta coloration; and because it is diastase-resistant, less brilliant than that of mucus but more so than bronchiolar cell secretions, and finer textured than lysosomal staining of other cells present, the effect is to highlight small-granule cells whether solitary or in clusters. Additional PAS staining of basement membranes and lead hematoxylin staining of cilia enhance the combined stain's resolving power. In thyroid gland, parafollicular cells stand out boldly against follicular elements; in small intestine, hematoxylin-positive endocrine cells are well differentiated from absorptive, mucous, and Paneth cells that surround them. Using a complementary monoamine fluorescence technique on plastic sections of lungs from control and 5-hydroxytryptophan-pretreated animals prior to staining, we can show that fluorescent epithelial cells are identical with those stained by PAS-lead hematoxylin.

Adrenal Medulla↗

PubMed1984

Acetylcholinesterase and choline acetyltransferase staining of neurons in the opossum esophagus.

The purpose of the present investigation was to identify and compare cholinergic intramural neurons in the lower esophageal sphincter and esophageal body by histochemical staining for acetylcholinesterase and the enzyme that synthesizes acetylcholine, choline acetyltransferase. Opossums were anesthetized and their abdominal cavity was opened by a midline incision to expose the esophagogastric junction. The lower esophageal sphincter was identified manometerically and localized in situ with markers. Tissues were removed, rapidly frozen in freon cooled with liquid nitrogen and serial cryostat sections were obtained from the lower esophageal sphincter and esophageal body. Sections were stained with one of the above histochemical procedures and adjacent sections were stained with Solachrome cyanin , which differentially stains nerve elements from muscle fibers. The muscle of the lower esophageal sphincter and esophageal body was stained with nonspecific cholinesterase with some selectivity of intensity of reaction in the various smooth muscle layers. All identifiable plexus neurons in the esophagus stained for nonspecific cholinesterase and acetylcholinesterase. Nerve fiber tracts were also stained for acetylcholinesterase within the longitudinal and circular layers of the tunica muscularis. Reaction for choline acetyltransferase showed no staining in the muscle layers or nerve fiber tracts of either part of the esophagus studied; however, selected neurons within the myenteric plexus of both regions (approximately 38%) were reactive. There was no significant difference in the number of positive choline acetyltransferase neurons in the lower esophageal sphincter or esophageal body.

Acetylcholinesterase↗

PubMed1990

Changes in glycoconjugates revealed by lectin staining in the developing airways of Syrian golden hamsters.

Lectin binding was studied in the developing airways of Syrian golden hamsters on gestational days 11-16 (day 16 is the day of birth). The trachea and lungs were fixed in 4% formaldehyde-1% glutaraldehyde, 6% mercuric chloride-1% sodium acetate-0.1% glutaraldehyde, and 95% ethanol; embedded in paraffin; and stained with eight lectin-horseradish peroxidase conjugates: Triticum vulgare (WGA), Dolichos biflorus (DBA), Helix pomatia (HPA), Maclura pomifera (MPA), Griffonia simplicifolia I-B4 (GSA I-B4), Arachis hypogaea (PNA), Ulex europeus I (UEA I), and Limulus polyphemus (LPA). Each lectin yielded a characteristic staining pattern, which modulated throughout development. In general, changes in staining characteristics of the tracheal epithelium preceded similar changes in the lobar bronchus, bronchiole, and alveolus. In the case of UEA I, MPA, WGA, and HPA, staining increased with time uniformly over the luminal surface of all epithelial cells. However, in the case of PNA, GSA I-B4, and LPA, after the differentiation of ciliated and secretory cells, the apical surfaces of the ciliated cells stained more intensely than the apical surfaces of the secretory cells. Neuraminidase pretreatment enhanced PNA and GSA I-B4 staining in both cell types. In the case of PNA, these light microscopic observations were confirmed by ultrastructural study. Unlike the other lectins, the pattern of staining with DBA was unusual. Staining was moderate at first, then decreased (days 13 and 14), then increased at all airway levels. This study shows that different glycoconjugates modulate in airway epithelial cells throughout fetal development.

Animals↗

PubMed1992

Area 3a in the cat. I. A reevaluation of its location and architecture on the basis of Nissl, myelin, acetylcholinesterase, and cytochrome oxidase staining.

Current knowledge on the anatomy of area 3a of the cat mainly derives from the cyto- and myeloarchitectonic study of Hassler and Muhs-Clement (J Hirnforsch 6:377, 1964). Previous investigations in the cat had failed to identify a cortical region comparable to monkey's area 3a. In the present study, Nissl, myelin, acetylcholinesterase, and cytochrome oxidase staining techniques were applied to coronal and sagittal serial sections of the cat brain. Area 3a appears as a slender band of cortex between areas 4 and 3b, and in Nissl-stained sections it is mainly characterized by an attenuated granular layer IV, overlying a thin layer V with pyramidal cells of various sizes, including a few large ones. These cytoarchitectonic features are sufficient to differentiate area 3a from neighboring areas, although the borders between them are not sharp in many cases. After the Nissl staining, the acetylcholinesterase staining proved to be the most helpful in defining the structure and borders of area 3a. Acetylcholinesterase staining was dense in layer I (in contrast with a lighter staining of outer layer I in area 4), and light in layers II and IIIa, changing to moderate in IIIc and IV (a pattern which is accentuated in area 3b). Myelin and cytochrome oxidase techniques also yielded differential staining patterns of area 3a and neighboring areas 4 and 3b, although the borders were not easily drawn with these techniques. Whereas our cyto- and myeloarchitectonic findings were comparable to those of Hassler and Muhs-Clement ('64) and applied well to area 3a in the convexity of the hemisphere, we found that most of the area 3a described by these authors in the medial face of the hemisphere had a number of distinguishing architectonic (as well as connectional and physiological) features which enabled us to define it as a separate area (7m). The techniques we used to delineate area 3a are compatible with most current procedures of histo- and immunohistochemical staining of the brain, and may also provide valuable supporting data for electrophysiological studies.

Acetylcholinesterase↗

PubMed1988

Flow cytometric detection of herpes simplex virus type 2 infected and transformed cells by immunoenzymatic and by indirect immunofluorescence staining.

The glucose oxidase antiglucose oxidase (GAG) immunoenzymatic staining procedure has been used to detect herpes simplex virus (HSV) antigens microscopically. In this study, the GAG procedure was adapted to cells in suspension, and its potential usefulness in flow cytometry was examined. HSV-2 infected monkey kidney and HSV-2 transformed mouse cells were stained using antisera to HSV-2 or to an HSV-2 specific protein with a molecular weight of 38 Kd, respectively, with the GAG procedure. Flow cytometric analysis of the GAG stained cells was then performed by the measurement of scattered light intensity in the angular intervals 1 degree-2 degrees, 2.5 degrees-19 degrees, and 3 degrees-6 degrees. The greatest scattered light intensity decrement caused by staining occurred in the 3 degrees-6 degrees angular interval, as predicted by previous work. In infected cells, which stain intensely by immunofluorescence, the difference between positively and negatively stained cells was adequate for detecting infected cells using the GAG method; however, this was not the case for the lightly staining transformed cells. The indirect immunofluorescence method of analysis of the same populations was superior to the scattered light method of analysis of the GAG stained infected and transformed cells.

Animals↗