[The centrosome of leukocytes of vertebrates studied by phase contrast microcinematography and by electron microscopy].
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Animals↗
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[Use of phase-contrast and interference-contrast microscopy for immediate cytological tests in gynaecological practice (author's transl)].
Vital cytological tests of cervical smears obtained from 1,083 patients in a gynaecological practice are reported. The smears had been taken in search for malignant changes, in addition to the need for appraisals of ovarian function and vaginal flora. Smears which produced suspicious colposcopic and vital cytological findings were restained by means of Papanicolaou stain. The diagnoses based on both methods were compared. The results have confirmed the vital cytological technique to be extremely suitable for functional and flora diagnosis, while the Papanicolaou method was found to be superior to vital cytology in cancer screening.
Cytodiagnosis↗
[Laboratory diagnosis of Acanthamoeba keratitis].
OBJECTIVE: To find a rapid method for diagnosing Acanthamoeba keratitis and identifying Acanthamoeba. METHODS: 10% potassium hydroxide (KOH) wet-mount preparations, Acanthamoeba culture, inverted phase contrast microscopy, and pathological examination using H. E. staining and PAS staining. RESULTS: Using corneal scrapings and corneal materials obtained from surgery, 7 cases and 5 cases of Acanthamoeba keratitis were diagnosed by 10% KOH wet-mount preparations. 6 strains of Acanthamoeba were isolated in corneal materials of 6 cases by protozoa culture method. The cysts, trophozoites and pseudopods on the trophozoites of Acanthamoeba were directly observed under the inverted phase contrast microscope. The cysts and trophozoites of Acanthamoeba were seen by H. E. staining and PAS staining with 20 h. CONCLUSION: Acanthamoeba keratitis could be rapidly diagnosed by 10% KOH wet-mount preparations and inverted phase contrast microscopy. Acanthamoeba organisms could be directly observed and identified under inverted phase contrast microscope.
Acanthamoeba↗
Morphology of spermatogenic arrest in man.
Spermatogenic arrest in man was studied by transillumination, phase contrast microscopy, and conventional histology. The spermatogenic arrest occurred at the late pachytene stage, and the degeneration of the germ cells could be studied more accurately with phase contrast microscopy of living cells than with conventional histology. The Sertoli cells rapidly phagocytosed the degenerating cells, which could be seen on transillumination as dark spots along the whole length of the tubule.
Follicle Stimulating Hormone↗
Modeling aberrant wound healing using tissue-engineered skin constructs and multiphoton microscopy.
BACKGROUND: Keloids and hypertrophic scars result from aberrant wound healing and remain a potential complication of any surgical procedure or trauma. Investigation of aberrant wound healing has been limited to the study of growth factors, collagen precursors, and DNA synthesis in simple in vitro systems, which necessitate removal or destruction of cells or factors in the growth environment of cell cultures. Multiphoton microscopy (MPM) can use endogenous chromophores such as collagen and nicotinamide adenine dinucleotide hydrogenase to produce thin optical sections of thick living tissues without the use of dyes or stains. Endogenous second-harmonic-generation (SHG) signals in collagen can be collected to form an MPM image. OBJECTIVE: To present a novel wound-healing model used to investigate keloid-derived fibroblast activity and collagen production in the same intact tissue-engineered construct over time. METHODS: Artificial tissue constructs called RAFTs (produced by suspension of keloid or normal dermal fibroblasts in type I collagen gel with an overlying keratinocyte layer) were cultured at air-fluid interface. Multiphoton microscopy SHG images of collagen in the intact tissue constructs consisting of normal or keloid-derived fibroblasts were obtained. The constructs were then incised with a scalpel. Serial MPM and phase-contrast microscopy images were obtained to monitor changes in the extracellular matrix in response to wounding of the artificial skin construct over 8 days. RESULTS: The tissue-engineered constructs formed a bilayer resembling the dermis and epidermis of human skin. Phase-contrast microscopy revealed migration of keratinocytes into the defect created by scalpel wounding. The constructs were found to contract with time after wounding. The MPM SHG images showed collagen deposition in the tissue constructs after wounding. Tissue constructs with keloid-derived fibroblasts were found to deposit collagen at a higher rate than those with normal fibroblasts. CONCLUSIONS: The MPM model described herein permits serial observation of the same intact specimens without the need for fixation or cytotoxic stains. Furthermore, it demonstrates the biologic activity of RAFT artificial tissue constructs.
Cells, Cultured↗
Scanning electron microscopy of L-phase streptococci. I. Development of techniques.
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Culture Media↗
Cell wall deficient bacteria as a cause of idiopathic hematuria.
Idiopathic hematuria in the absence of bacteriuria is a medical challenge. Routine cultures of catheterized bladder and endoscopically obtained ureteral urine specimens from a 22-year-old woman with a 6-week history of hematuria showed no growth after 24 to 48 hours of incubation. However, bacterial variants were grown on enriched media. Colonies were typical cell wall deficient/defective bacteria. Phase and electron microscopy of cystoscopic urine specimens obtained by retrograde ureteral catheterization as well as phase microscopy of the cultures revealed the classic morphology of these organisms. When the variant cultures were subcultured the organisms reverted to their related walled forms, that is Streptococcus agalactiae and Staphylococcus haemolyticus. Because the colonies of these organisms showed various patterns of biochemical reactivity, each phenotype was tested against 15 antimicrobials. Collectively, all biotypes had a common susceptibility to only nitrofurantoin. The patient was treated with nitrofurantoin for 6 weeks. Four days after initiation of therapy she had complete remission of hematuria. During the next 3 years she remained well and free of hematuria.
Adult↗
Multinuclear NMR microscopy of two-phase fluid systems in porous rock.
The high-field magnetic resonance (MR) characteristics of fluids in porous reservoir rock exhibit short T2 relaxation times and broad natural line widths. These characteristics severely restrict which MR imaging (MRI) methodology can be used to obtain high-resolution porescale images of fluids in porous rock. An MR microscopy protocol based on 3D backprojection using strong imaging gradients was developed to overcome many of these constraints. To improve the image quality of two-phase systems, multinuclear MRI using proton MR to image the brine phase and 19F MR of a fluorinated hydrocarbon to image the oil phase was used. Resolution as high as 25 microns per pixel has been obtained for fluid systems in Bentheim and Fontainebleau sandstones. Separate proton and 19F images of brine and oil phases show good agreement with total saturation images. Software has been developed to perform 3D erosion/dilations and to extract the pore size distribution from binarized 3D images of fluid filled porosity. Results from pore size measurements show significant differences in the nature of the pore network in Fontainebleau and Bentheim sandstones.
Image Processing, Computer-Assisted↗
Comparison of fluorescence microscopy and solid-phase cytometry methods for counting bacteria in water.
Total direct counts of bacterial abundance are central in assessing the biomass and bacteriological quality of water in ecological and industrial applications. Several factors have been identified that contribute to the variability in bacterial abundance counts when using fluorescent microscopy, the most significant of which is retaining an adequate number of cells per filter to ensure an acceptable level of statistical confidence in the resulting data. Previous studies that have assessed the components of total-direct-count methods that contribute to this variance have attempted to maintain a bacterial cell abundance value per filter of approximately 10(6) cells filter(-1). In this study we have established the lower limit for the number of bacterial cells per filter at which the statistical reliability of the abundance estimate is no longer acceptable. Our results indicate that when the numbers of bacterial cells per filter were progressively reduced below 10(5), the microscopic methods increasingly overestimated the true bacterial abundance (range, 15.0 to 99.3%). The solid-phase cytometer only slightly overestimated the true bacterial abundances and was more consistent over the same range of bacterial abundances per filter (range, 8.9 to 12.5%). The solid-phase cytometer method for conducting total direct counts of bacteria was less biased and performed significantly better than any of the microscope methods. It was also found that microscopic count data from counting 5 fields on three separate filters were statistically equivalent to data from counting 20 fields on a single filter.
Biomass↗
A rapid method for identification of S-phase cells in transmission electron microscopy of monolayer cultures.
A rapid technique has been developed for identifying cells in S-phase in transmission electron microscopy of monolayer cultures. Autoradiography is performed on the embedded monolayer which is then re-embedded. The cells and the adjacent layer of developed emulsion are sanctioned together, and when subsequently viewed in the electron microscope the cells in S-phase appear with a zone of silver grains in the emulsion which is opposite their nuclei. They are thus readily identified and can be selected for detailed study. The advantages of this technique over conventional TEM autoradiography are the very much shorter exposure times and a reduced workload when preparing serial sections.
Cells, Cultured↗
Design of a capacitively decoupled transmit/receive NMR phased array for high field microscopy at 14.1T.
A design is presented for a "phased array" of four transmit/receive saddle-geometry volume coils for microimaging at 600 MHz within a 45 mm clear-bore vertical magnet. The small size of the coils, approximately 10 mm in length, and high frequency of operation both present considerable challenges for the design of a phased array. The particular design consists of four saddle coils, stacked vertically, in order to produce an array suitable for imaging samples, typical of many microimaging studies, with a large length:diameter ratio. Optimal coil overlap is used to reduce the mutual inductance between adjacent coils, and capacitive networks are used to maximize the isolation between all of the coils. Standard 50 Omega input impedance preamplifiers are used so that the preamplifiers do not have to be integrated directly into the probe. Isolation between coils was better than 20 dB for all coil pairs. An increase in signal-to-noise of 70 +/- 3% was achieved, averaged over the whole array, compared to a single coil of the same dimensions. High resolution phased array images are shown for ex vivo tissue samples.
Animals↗
[Albumin receptors on isolated human hepatocytes. Studies by phase contrast and scanning electron microscopy].
In the present work a minibead method has been employed to study human hepatocyte receptors for albumin-coupled latex particles. The cell-latex reaction was observed in both phase contrast and scanning electron microscopy. It has been demonstrated that human hepatocytes exhibit bindings sites for different forms of glutaraldehyde-treated albumin, and the ligand has proved to be species-nonspecific. The albumin binding activity is temperature-dependent and can be only partially blocked by preincubation with free, glutaraldehyde-treated protein, unless hepatocytes are pretreated with vinblastine sulfate. The possible biological role of these receptors as regards the infection due to the hepatitis B virus (that shares common, but not identical binding sites) is also discussed.
Humans↗
Haematuria: glomerular or non-glomerular?
The standard urinary sediment of 80 consecutive patients with significant haematuria admitted at our hospital was examined for significantly dysmorphic red blood cells by 3 methods-1. Phase contrast microscopy, 2. Wright's staining of the urinary sediment, and 3. Simple light microscopy. The results of the study were compared with the final diagnosis reached in the ward and the sensitivity of the three methods was statistically compared. Our study conclusively proves that phase contrast microscopy is superior to light microscopy of plain or Wright's stained urinary sediment, with respect to both sensitivity and percentage of dysmorphism of the urinary red blood cells which can be detected.
Diagnosis, Differential↗
Scanning electron microscopy of L-phase streptococci. II. Growth in broth and upon millipore filters.
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Cell Membrane↗
The influence of surface-free energy on supra- and subgingival plaque microbiology. An in vivo study on implants.
The influence of surface free energy on supra- and subgingival plaque microbiology was examined in 9 patients with functional fixed prostheses supported by endosseous titanium implants. Two abutments (trans-mucosal part of the 2 stage implant) were replaced by either a new titanium abutment or a fluor-ethylene-propylene (FEP) coated abutment per subject. After 3 months of habitual oral hygiene, plaque samples were taken. Supragingival plaque was examined by means of differential phase-contrast microscopy, whereas for the subgingivial plaque additional analyses (DNA probes analysis, culturing) were performed. The subgingival samples were taken by paper-points and by scraping of the subgingival abutment surface. Differential phase-contrast microscopy showed a significant difference in plaque composition, especially when supragingival plaque was considered (P = 0.05). FEP coated abutments frequently harbored more coccoid microorganisms, whereas spirochetes or motile organisms were only detected around titanium abutments. Subgingivally, the number of colony forming units (CFU) in paper-points was comparable for both types of abutments. If the to-the-abutment-adhering plaque was considered, the number of CFU was 5 times higher on the titanium abutments than on the FEP coated abutments. However, this difference did not reach a statistical level of significance (P = 0.38). The DNA probe analysis of the subgingival plaque collected with paper-points showed a slightly higher frequency and concentration of perio-pathogens around the titanium abutments. However, the inter-substratum differences were smaller than the inter-subject differences. The latter seemed to be related to patient's dental status.(ABSTRACT TRUNCATED AT 250 WORDS)
Bacterial Adhesion↗
Liposomes from ionic, single-chain amphiphiles.
In studies of the minimum physiochemical requirements for lipid membrane formation, we have made liposomes from dilute, aqueous dispersions of C(8)-C(18) single-chain amphiphiles. In general, membrane formation from ionic soaps and detergents requires the presence of uncharged amphiphiles. Vesicles were characterized by phase-contrast microscopy, by trapping of ionic dyes, as well as by negativestain and freez-frature electron microscopy. They were typically heterogeneous in size, but the average diameter could be experimentally varied in some cases over the range of 1 to 100 micrometer. Uni-, oligo-, and multilamellar vesicles were observed. Membrane permeability to various solutes was determined in part by a new technique which utilized phase-contract microscopy; when impermeable vesciles exclude added solutes such as sucrose, refractive index differences are created between vesicle contents and surrounding medium, so that the vesicles appear bright in the phase microscope. Permeant solutes do not produce this effect. Spectrophotometric permeability determinations confirmed the results of this technique and provided quantitative measures of permeability. Monoalkyl liposomes have potential uses as models of biomembranes and in drug delivery. They are also relevant to the prebiotic origin of biomembranes.
Fatty Acids, Nonesterified↗
The genus Candida Berkhout. XI. Electron microscopy and roentgen-phase analysis of cell walls of three strains of Candida guilliermondii (Cast.) Langeron et Guerra.
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Agglutination Tests↗
[Differential diagnosis of urinary findings].
Classical urinalysis is extremely useful in the differential diagnosis of diseases of the kidneys and lower urinary tract. Although dipsticks are suitable for orientational purposes, when pathology is indicated, and when renal disease presents, they must be supplemented by urine microscopy or quantitative and qualitative protein electrophoresis to establish the diagnosis. Phase-contrast microscopy can distinguish glomerular from nonglomerular hematuria and thus guide the further diagnosis, and the detection of leukocytes and bacteria confirms the diagnosis of urinary tract infection. In asymptomatic patients microhematuria or proteinuria is detected more frequently with increasing age. A differentiated diagnostic strategy must be adapted to the individual risk of the patient, and must avoid overdiagnosis while not missing potentially serious pathology.
Acanthocytes↗