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At least 1,297 records · Page 72Linked to original sources
PubMed1998

Introduction: electron microscopy for fungal cell ultrastructure

Ultrastructural morphology is a principal study for almost all natural science, i.e., an inevitable and fundamental study to elucidate structures, functions and differentiation of a given tissue, cell, or molecule as a most understandable visual form. The number of young scientists, however, who have been engaged in this field is very small recently, since this field appears to be too old when compared with modern molecular biology and it takes much longer time for the beginner to master the methology for electron microscopy (EM). This symposium is designed for these young scientists and molecular biologists or biochemists, who are not so familiar to ultrastructural morphology, to better understand the applicability of EM, through new results and findings in ultrastructures of fungal cells and related organisms. EM includes several kinds of methods, which are shadowing EM, negative staining EM, ultrathin section EM, scanning EM, freeze-fracture EM, immuno-EM and diverse methods for staining the specimen. Shadowing EM and negative staining EM are suitable methods for the study molecular structures of proteins, and the former is prepared by shadowing with platinum palladium in a vacuum chamber, and the latter is a method to observe a relief prepared by dipping the sample in phosphotungsten solution. Freeze-fracture electron microscopy is suitable for the study of membrane plane ultrastructures, since it reveals a wide planar view of the membrane by splitting it along the hydrophobic membrane internal plane. Immunoelectron microscopy is an essential method for the study of intracellular localization of proteinous molecules. These methods will be introduced. This symposium will introduce new findings as for fungal cells, bacteria and protozoa obtained principally by using electron microscopy. These findings obtained through ultrastructures may provided a renewed knowledge of research approach from view points of ultrastructure.

Journal Article↗
PubMed2001

Comparison of a nested polymerase chain reaction--restriction fragment length polymorphism method, the PATH antigen detection method, and microscopy for the detection and identification of malaria parasites.

A nested polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method, the PATH antigen detection method, and light microscopy were compared for their capacity to detect and identify Plasmodium species. One hundred and thirty-six blood specimens obtained from patients suspected of having malaria were examined by each of the three methods. Forty-four specimens were positive for malaria using microscopy as the "gold standard". The sensitivity for nested PCR was 100%, and the specificity was 98%. For the detection of Plasmodium falciparum, the antigen detection method had a sensitivity of 100% and a specificity of 97%. Species identification obtained using PCR-RFLP was identical or superior to light microscopy in 42 cases (96%). Although the nested PCR-RFLP method was more sensitive and specific, the rapid turnaround time and high sensitivity of the antigen detection method makes it a useful adjunct to standard microscopy.

Animals↗
PubMed2002

Quality assessment of sputum transportation, smear preparation and AFB microscopy in a rural district in Malawi.

SETTING: Ntcheu District, Central Region of Malawi. OBJECTIVES: To assess 1) the feasibility of introducing simple internal quality control procedures for acid-fast bacilli (AFB) microscopy, and 2) the quality of the district sputum smear microscopy service. DESIGN: A simple internal quality control system was piloted in which district laboratory staff assessed: 1) specimen suitability, 2) time between sputum submission and smear examination, 3) smear preparation and staining, and 4) microscopy. Actual times for processing specimens were compared with recommended times. External quality validation was carried out. RESULTS: Of 4805 sputum specimens: 1) documentation was complete in 95%, 2) 93% reached the laboratory within 7 days of collection, 3) 96% of smears were well prepared and stained, and 4) 97% concordance (96.4% smear-positive and 97.6% smear-negative) was demonstrated when 208 smears were re-examined by a second technician. The aggregate index of reliability was 86%. The mean time spent on microscopic examination was 3.8 minutes, compared with the recommended time of 10 minutes. When all smears from 164 patients were assessed externally, 98.2% concordance (98.1% smear-positive and 98.2% smear-negative) was demonstrated. False smear-negative and smear-positive rates were less than 2% each. CONCLUSION: District laboratory staff were able to incorporate simple quality control procedures for AFB microscopy into their routine practice, resulting in a reliable service. The lessons learnt are widely relevant and potentially useful for implementation of a national quality assurance scheme.

Chi-Square Distribution↗
PubMed1992

[Application of electron microscopy in the diagnosis of neoplasms].

The majority of neoplasms can be diagnosed by light microscopy, but in some cases the diagnosis remains ambiguous due to poor differentiation, even though special stainings have been employed. This paper presents 34 cases of neoplasms in which the tumors were diagnosed by electron microscopy. This includes distinguishing (1) anaplastic carcinoma from lymphoma; (2) anaplastic carcinoma from amelanotic melanoma; (3) APUDoma from other tumors; (4) different mesenchymal tumors. The diagnoses of 4 cases of malignant melanoma, 11 cases of APUDoma, 7 cases of poorly differentiated carcinoma or anaplastic carcinoma, 2 cases of non-Hodgkin's lymphoma, 9 cases of mesenchymal tumors and 1 cases of other tumors have been resolved by electron microscopy. It is obvious that in some cases, electron microscopy can be of help in establishing a correct diagnosis.

Apudoma↗
PubMed1992

The validity of frozen section diagnosis based on video-microscopy.

To investigate the accuracy of video-microscopy of frozen sections, two pathologists reexamined 80 cases of archival material, on which frozen sections had previously been performed. Diagnoses based on the two trials (Observer 1 and 2) and diagnoses obtained in the original frozen section situation were compared with the final diagnoses based on paraffin embedded material. Observer 1 had two false negative diagnoses, but no false positive, whereas Observer 2 had one false positive, but no false negative diagnosis when compared with the final diagnoses. No false positive or false negative diagnoses were made in the original frozen sections situation and the number of inconclusive diagnoses were 5, as compared with 6 and 8 in the two trials based on video-microscopy. More experience with video-microscopy will probably achieve a quality of frozen section diagnoses similar to that of direct light-microscopy.

Diagnosis, Differential↗
PubMed1992

Capillary microscopy in patients with vasospastic and arterial occlusive diseases.

Capillary microscopy and its application for diagnosis and therapeutic evaluation in patients with vasospastic and arterial occlusive disease are described in this report. Capillary microscopy is a noninvasive technique for measuring red blood cell velocity and for determining nutritional blood flow in the capillaries of the skin. This method can be applied in distinguishing patients with Raynaud's phenomenon from those with other disorders involving ischemia of the hand. In patients with atherosclerotic disease of the lower limb, microcirculatory observations can help determine how temperature increases and posturally induced microvascular constriction affect limbs with different degrees of ischemia. Capillary microscopy can also be used to help distinguish between patients who respond and those who do not respond to prolonged treatment for such disorders. Because capillary microscopy can be used to assess microcirculatory hemodynamics and autoregulatory mechanisms in cases of severe ischemia, it should be regarded as an essential tool in the evaluation of patients with upper- and lower-limb ischemia.

Journal Article↗
PubMed2005

Can the urine dipstick test reduce the need for microscopy for assessment of systemic lupus erythematosus disease activity?

OBJECTIVE: Urine microscopic examination is an important component of the Systemic Lupus Erythematosus Disease Activity Index (SLEDAI). We investigated whether the urine dipstick test can reduce the need for microscopy for the assessment of SLEDAI. METHODS: We studied 269 urine samples from 259 SLE patients with Albustix and Hemastix reagent strips. The results were compared to concomitant microscopic examination of urinary sediment. RESULTS: When trace red blood cell was defined as the cutoff, the sensitivity, specificity, and negative predictive value (NPV) of the Hemastix urine test were 0.98, 0.53, and 0.99, respectively, for hematuria; 0.82, 0.47, and 0.90, respectively, for the presence of pyuria; and 0.91, 0.44, and 0.98, respectively, for the presence of casts by microscopic examination. When proteinuria of 1+ was defined as the cutoff, the sensitivity, specificity, and NPV of the Albustix test were 1.00, 0.46, and 0.99, respectively, for urinary casts; and 0.82, 0.49, and 0.90, respectively, for the presence of pyuria. When both Albustix and Hemastix were applied as screening test, urine microscopy could be reduced by 27%; however, 8% of cases with normal Albustix and Hemastix tests had at least one abnormality on urine microscopy examination. CONCLUSION: In patients with SLE, a combination of Albustix and Hemastix urine tests showed reasonable sensitivity to detect abnormalities in urine sediment. Based on these results, routine urine microscopy can be limited to SLE patients with abnormal Albustix or Hemastix tests. Rarer causes of abnormal renal function in lupus, such as tubulointerstitial nephritis or drug induced interstitial nephritis, would be manifested by pyuria and therefore would not necessarily be detected by changes in the blood and protein detectors on the urine dipstick.

Adolescent↗
PubMed1976

Scanning electron, light, and immunofluorescent microscopy of coronaviral enteritis of turkeys (Bluecomb).

Intestinal sections from both experimental and field cases of turkey coronaviral enteritis (TCE) were examined by scanning electron microscopy and light microscopy through 10 days after inoculation and by a direct fluorescent antibody test for TCE through 12 days. Serums were collected for an indirect fluorescent antibody test for TCE through 160 days after inoculation. Lesions observed with the scanning electron microscopy were catarrhal enteritis with hemorrhage per diapedesis, epithelial desquamation, and villous atrophy which developed and regressed within 6 days after inoculation. Light microscopy demonstrated similar lesions, except that villus-to-crypt ratios remained depressed 10 days. The direct fluorescent antibody test demonstrated the presence of coronaviral antigen throughout the sampling period, and serum antibodies to TCE were present until at least 160 days, when the experiment was terminated.

Animals↗
PubMed1990

Microhaematuria in general practice: is urine microscopy misleading?

During a health centre screening programme for men aged 60 years and over, the urinary dipstick results of 58 patients found to have microscopic haematuria were compared with the results of immediate microscopy at the health centre and routine microscopy by a hospital laboratory. There was agreement between a positive dipstick test for haematuria and the presence of red cells at routine microscopy reported by the hospital laboratory in only 18 cases (31%). Routine urine microscopy results requested from general practice should not be taken as the criterion for deciding whether further investigation is needed in cases of microscopic haematuria.

Aged↗
PubMed1979

Ultrastructural pathology with normal light microscopy of liver in Wilson's disease. A case report.

Light microscopy findings in early Wilson's disease (hepatolenticular degeneration) may be normal even when special cytochemical stains are used. We present a case of Wilson's disease in which light microscopy was negative, while electron microscopy showed the characteristic changes in the hepatocytes. Since low serum ceruloplasmin levels and high urinary copper excretion are not by themselves definite proof of Wilson's disease, and since copper content of the liver is not universally measured, electron microscopy examination of liver tissue appears to be a worthwhile additional tool for the early diagnosis of Wilson's disease.

Adult↗
PubMed1979

Urine microscopy and infection in general practice.

To test the value of urine microscopy 100 consecutive specimens were examined in the surgery and the results correlated with the subsequent laboratory culture reports.An assessment of the degree of pyuria was made by low power microscopy of a thick drop of fresh urine. A second specimen was examined under high power for the presence or absence of motile bacilli. The techniques used are described and quantified.The laboratory report was definitive in 88 of the 100 cultures. All the 33 specimens with 10(5) bacteriuria had some degree of pyuria and in 27 (82 per cent) motile bacilli had been found. In the 50 with no significant bacteriuria no motile bacilli had been seen in 38 (76 per cent).In these 88 specimens a diagnosis made in the surgery based entirely on bacterial microscopy would have been correct in 80 per cent, combined with cytological microscopy in 87 per cent, and with the addition of clinical features in 92 per cent.In the remaining 12 cases the laboratory report was inconclusive and would have made no difference to my conclusions.

Bacteriuria↗
PubMed1980

Ultrastructural and elemental imaging of biological specimens by soft X-ray contact microscopy.

Soft X-ray contact microscopy offers a means of visualizing unstained as well as stained biological materials at better than 6 nm resolution. Unlike light and transmission electron microscopy, which rely on stains to increase specimen contrast, soft X-ray imaging is dependent upon the differential absorption of incident soft (1-10nm wavelength) X-rays by the endogenous elements within the specimen. The advantages of using soft X-rays for imaging are: 1) reduced specimen damage during exposure; 2) ability to image hydrated specimens at atmospheric pressure; 3) ability to image specimens ranging in thickness from less than 40 nm to as much as 10 microns; and 4) ability to map the elemental composition of the specimen through observation of the differential absorption of properly chosen incident x-ray wave lengths. This paper explains the principles of the image formation and demonstrates the use of soft X-ray contact microscopy with biological samples which could not readily be imaged in their natural form using conventional electron microscopy methods.

Animals↗
PubMed1980

Scanning electron microscopy of the adult human testis.

Scanning electron microscopy was performed on adult human testicular tissue using fresh orchiectomy and some recent autopsy material. After preliminary fixation in 2.5% glutaraldehyde in phosphate buffer, pH 7.4, blocks were taken for scanning electron microscopy and for correlative light and transmission electron microscopy,. For scanning electron microscopy, the tissue was processed by means of the osmium-thiocarbohydrazide-osmium method, dehydrated and critical point dried with CO2. Blocks taken from the periphery of the testis showed the presence of the tunica albuginea, which formed a comparatively dense encapsulation around the testis. Large blood vessels were present at points along the innermost layer of the tunica. Within the testis, the vessels became progressively smaller and came into close apposition to the seminiferous tubules. Individual tubules could be identified from the lowest magnification (15x). They were seen both as intact, intricately convoluted structures and at all levels and planes of transection. Within the tubules, the seminiferous epithelium was seen both in the intact and transected state. All of the cell types, were identified. In the basal layer, in contact with the basement membrane, there were spermatogonia and varying numbers of Sertoli cells. Spermatocytes were located in the mid-zone of the tubule and spermatids in the adluminal compartment. There were numerous tails of spermatozoa projecting into the lumen. Normal spermatozoa, some round headed forms and an occasional bizarre multiheaded spermatozoon were observed. Leydig cells were noted in the intertubular angles and in close proximity to blood vessels.

Adult↗
PubMed1984

Coalescence of endothelial cells in the traumatized cornea. III. Correlation between specular and scanning electron microscopy.

Sequential specular microscopy has shown that coalescence of endothelial cells occurs after thawing in cryopreserved rabbit corneas. Scanning electron microscopy of corneas studied in vitro by specular microscopy showed the same large, fused endothelial cells. As observed in the electron micrographs, these cells showed no evidence of the cell borders that had disappeared during the specular microscopic examination. These data support the reliability of our specular microscopic observations and provide additional evidence for the occurrence of fusion or coalescence of corneal endothelial cells.

Animals↗
PubMed1998

Pre-embedding immunolabeling for electron microscopy: an evaluation of permeabilization methods and markers.

For scarce antigens or antigens which are embedded in a dense macromolecular structure, on-section labeling, the first method of choice, is not always successful. Often, the antigen can be localized by immunofluorescence microscopy, usually by a pre-embedding labeling method. Most of these methods lead to loss of ultrastructural details and, hence, labeling at electron microscope resolution does not add essential information. The scope of this paper is to compare five permeabilization methods for pre-embedding labelling for electron microscopy. We aim for a method that is easy to use and suitable for routine investigations. For our ongoing work, special attention is given to labeling of the cell nucleus. Accessibility of cytoplasmic and nuclear antigens is monitored with a set of different marker antibodies. From this investigation, we suggest that prefixation with formaldehyde/glutaraldehyde is necessary to stabilize the ultrastructure before using a detergent (Triton X-100 or Brij 58) to permeabilize or remove the membranes. The experimental conditions for labeling should be checked first with fluorescence or fluorescence-gold markers by fluorescence microscopy. Then either ultrasmall gold particles (with or without fluorochrome) with silver enhancement or, if the ultrasmall gold particles are obstructed, peroxidase markers are advised. The most promising technique to localize scarce antigens with good contrast is the combination of a pre-embedding peroxidase/tyramide-FITC or -biotin labeling followed by an on-section colloidal gold detection.

Cell Nucleus↗
PubMed1999

Microscopy for recognition of individual biomolecules.

One frontier challenge in microscopy and analytical chemistry is the analysis of soft matter at the single molecule level with biological systems as most complex examples. Towards this goal we have developed two novel microscopy methods. Both employ highly specific molecular recognition schemes used by nature-the recognition of specific protein sites by antibodies and ligands. One method uses fluorescence labeled ligands for detecting single molecules in fluid systems like membranes (Fig. 1B). Unitary signals are reliably resolved even for millisecond illumination periods. The knowledge of the unitary signal from single molecules permits the determination of stoichiometries of component association (Fig. 3). Direct imaging of the diffusional path of single molecules became possible for the first time (Fig. 4). Using linear polarized excitation, the angular orientation of single molecules can be analyzed (single molecule linear dichroism, (Fig. 5), which opens a new perspective for detecting conformational changes of single biomolecules. In the other method, an antibody is flexibly linked to the tip of an atomic-force microscope. This permits the identification of receptors in multi-component systems. Molecular mapping of biosurfaces and the study of molecular dynamics in the ms to s range become possible with atomic force microscopy.

Antibodies↗
PubMed1999

An evaluation of two-photon excitation versus confocal and digital deconvolution fluorescence microscopy imaging in Xenopus morphogenesis.

The ability to visualize cell motility occurring deep in the context of opaque tissues will allow many currently intractable issues in developmental biology and organogenesis to be addressed. In this study, we compare two-photon excitation with laser scanning confocal and conventional digital deconvolution fluorescence microscopy, using the same optical configuration, for their ability to resolve cell shape deep in Xenopus gastrula and neurula tissues. The two-photon microscope offers better depth penetration and less autofluorescence compared to confocal and conventional deconvolution imaging. Both two-photon excitation and confocal microscopy also provide improved rejection of "out-of-focus" noise and better lateral and axial resolution than conventional digital deconvolution microscopy. Deep Xenopus cells are best resolved by applying the digital deconvolution method on the two-photon images. We have also found that the two-photon has better depth penetration without any degradation in the image quality of interior sections compared to the other two techniques. Also, we have demonstrated that the quality of the image changes at different depths for various excitation powers.

Animals↗
PubMed1997

Quantitation of cell-matrix adhesion using confocal image analysis of focal contact associated proteins and interference reflection microscopy.

We have developed an approach for the quantitation of vinculin, a focal contact associated protein, based on a multimodal confocal microscopy and image analysis. Vinculin spot distribution was imaged in confocal fluorescence microscopy and the corresponding focal contacts were imaged in confocal interference reflection microscopy. These images were analyzed with a SAMBA image cytometer. The image analysis program provided 12 morphometric features describing cellular area, shape, and proportions of vinculin spots as well as six topographical features describing the distribution of vinculin and the relative overlap of vinculin and focal contacts. This approach was applied to the study of rat osteosarcoma cells submitted to mechanical stresses: successions of 2g and 0g accelerations during a series of parabolic flights. The measured features were assessed by means of correlation analysis and stepwise discriminant analysis. After correlation analysis, only ten parameters were retained. Quantitation of cell morphological parameters indicated that cell area was significantly affected by gravitational stresses as well as vinculin distribution. Cell area was reduced by 50% and vinculin spots were restricted to cell periphery. Cell adhesion measured by IRM decreased significantly in the first part of the flight and remained stable at the end of the flight. These results suggest that cell-matrix adhesion is affected by gravitational stresses. Image analysis provides useful tools to investigate focal adhesion re-organization under different physiological stimuli.

Animals↗