The technique of electron microsco-pic in situ hybridization is applying in situ hybridization at the electron microscopic level. It is mainly used in the ultrastructural localization of the lablled DNA, RNA and RHA in a cell and/or a tissue. In this paper I mainly elaborated its establishment and classification, and the operation procedure of nonradioactive electron microscopic in situ hybridization and some points for attention. In the end I also discussed its application for research.
In Situ Hybridization ; methods ; Microscopy, Electron ; methods

Microscopy, Electron, Transmission ; methods ; Negative Staining ; methods ; Viruses ; ultrastructure

This article introduces the basic theories about atomic force microscope (AFM) and electron microscope (EM), respectively. New applications of each microscopic technology in regenerative medicine, covering both material science and life science, are discussed. The advantages or disadvantages of the kinds of microscopes in working conditions, sample preparation, resolution and the like, are discussed and compared systematically to make clear each scope of applications. This could be a useful guide for selecting the appropriate microscopic analysis in research work about regenerative medicine.
Humans ; Microscopy, Atomic Force ; methods ; trends ; Microscopy, Electron ; methods ; trends ; Regenerative Medicine ; methods ; trends

Confocal laser scanning microscopy (CLSM) was used to examine archaeoparasitological specimens from coprolites associated with La Cueva de los Muertos Chiquitos (CMC) located near present-day Durango, Mexico. The eggs for 4 different types of parasites recovered from CMC coprolites were imaged using CLSM to assist with identification efforts. While some of the parasite eggs recovered from CMC coprolites were readily identified using standard light microscopy (LM), CLSM provided useful data for more challenging identifications by highlighting subtle morphological features and enhancing visualization of parasite egg anatomy. While other advanced microscopy techniques, such as scanning electron microscopy (SEM), may also detect cryptic identifying characters, CLSM is less destructive to the specimens. Utilizing CLSM allows for subsequent examinations, such as molecular analyses, that cannot be performed following SEM sample preparation and imaging. Furthermore, CLSM detects intrinsic autofluorescence molecules, making improved identification independent of resource and time-intensive protocols. These aspects of CLSM make it an excellent method for assisting in taxonomic identification and for acquiring more detailed images of archaeoparasitological specimens.
Eggs ; Methods ; Mexico ; Microscopy ; Microscopy, Confocal ; Microscopy, Electron, Scanning ; Ovum ; Parasites

OBJECTIVETo observe the ultrastrual changes of the skin between the continuous tissue expansion and the conventional tissue expansion.

METHODSTwelve white piglets were used for this study on an animal model of tissue expansion with the continuous tissue expansion group and the conventional tissue expansion group. The tissue samples in each group were harvested and prepared for the transmission electron microscope observation.

RESULTSThe interspace among basal cells and spinose cells was increasing and the numbers of cell conjunctures were decreasing in the both groups. However, these changes in the continuous tissue expansion group were more obvious than in the conventional tissue expansion group. In the dermal layer of the skin, the ultrastructure of collagen fibers were basically normal. But, the fibroblasts and capillary endothelia cells were more activated in the continuous tissue expansion group, compared with the conventional tissue expansion group. The fibroblast apoptosis and collagenolysis spots were observed in both of the groups, while the red blood cells were also found in the tissue leaked outsides from the blood vascular cavities.

CONCLUSIONTissue expansion may result in tissue growth and tissue degeneration in the same time.

Animals ; Microscopy, Electron ; Skin ; cytology ; ultrastructure ; Swine ; Tissue Expansion ; methods

OBJECTTo study the significance of the ultra-high power microscope in the examination of vaginal discharge.

METHODSBy the ACT-2000 ultra-high power microscope system and Olympus CX21 microscope, the vaginal discharge of 1,100 gynaecology out-patients was examined respectively.

RESULTSThe positive rate of mould in the patients was 11.55% by CX21 and was 20.27% by ACT-2000, respectively. The positive rate of trichomonas vaginalis was 2.55% by CX21 and 3.0% by ACT-2000, respectively. The clue cell was detected in 11.27% of the patients by ACT-2000, but no such cell reported by CX21. Totally, positive results were obtained in 14.09% of the patients by CX21 and 32.55% by ACT-2000.

CONCLUSIONBy using the ultra-high power microscope, the positive result can be increased obviously in the examination of vaginal discharge. It is very important in clinical practices.

Adult ; Female ; Humans ; Microscopy ; methods ; Microscopy, Electron ; methods ; Middle Aged ; Vaginal Discharge ; pathology ; Young Adult

OBJECTIVETo study the histology and ultrastructure of the tidemark in the adult condylar cartilage and their significance.

METHODSAfter embedded in paraffin, 50 adult condyles were stained with HE, partly with Van-Gieson and histochemical methods, then observed by light microscope. 3 cases of the tidemark region were observed by transmission electron microscope, another 5 cases were studied by scanning electron microscope.

RESULTSIn the tidemark region, there had shown the presence of the AKP and calcium, absence of the proteoglycan, abundance of the membrane-bound matrix vesicles, crystals of hydroxyapatite and lipid nodule-like substances, which were often observed in the load-bearing areas. The collagen fibrils of the noncalcified cartilage crossed the tidemark gradiently and were continuous with those of the calcified cartilage; a band of horizontal fibrils surrounded the whole tidemark region, which was wider in the load-bearing areas than that in the nonload-bearing areas and which interweaved with the gradient fibrils so as to form a net. Digested with papain, the surface of the tidemark was highly undulating, and a lot of chondrocyte lacunae were seen on the surface, which were surrounded by calcified tissues.

CONCLUSIONSIn the region of the tidemark, physiological calcification takes place and is more active in the load-bearing areas; gradient and horizontal fibrils interweave with each other, which is correlated with the force on the articulation.

Adult ; Female ; Histocytochemistry ; methods ; Humans ; Male ; Mandibular Condyle ; cytology ; ultrastructure ; Microscopy, Electron ; Microscopy, Electron, Scanning ; Temporomandibular Joint ; cytology ; ultrastructure

OBJECTIVETo compare the efficacy of three methods for forensic diatom test, namely strong acid digestion-centrifuge enrichment-light microscopy (SD-CE-LM), microwave digestion-membrane filtration-automated scanning electron microscopy (MD-ME-SEM), and microwave digestion-membrane filtration-light microscopy (MD-MF-LM).

METHODSSixty samples were randomly divided into 3 groups for diatom test using three methods, and the sample preparation time, degree of digestion and recovery rate of diatoms were compared.

RESULTSThe sample preparation time was the shortest with MD-MF-LM and the longest with SD-CE-LM (P<0.05). MD-ME-SEM and MD-MF-LM allowed more thorough tissue digestion than SD-CE-LM. MD-ME-SEM resulted in the highest total recovery rate of diatom, followed by MD-MF-LM and then by SD-CE-LM (P<0.05); the recover rate of different diatom species was the highest with MD-ME-SEM, followed by MD-MF-LM and SD-CE-LM (P<0.05).

CONCLUSIONSD-CE-LM has a low recovery rate of diatoms especially for those with lengths shorter than 40 µm or densities less than 1/5. With a high recovery rate and accuracy in diatom test, MD-ME-SEM is suitable for diagnosis of suspected drowning cases. MD-MF-LM is highly efficient, sensitive and convenient for forensic diatom test.

Centrifugation ; Diatoms ; isolation & purification ; Drowning ; Forensic Sciences ; methods ; Humans ; Microscopy ; Microscopy, Electron, Scanning ; Microwaves ; Specimen Handling

Sterigmatocystin (ST), the secondary metabolite of many kinds of filamentous fungi, is a potent carcinogen structurally related to the aflatoxins (AFT). With similar chemical structure, sterigmatocystion behaves much the homogeneous properties to aflatoxins, both of these mycotoxins exhibit similar biological properties due to their bisfuranoid structure. Since the common, and even heavier pollution, found in foods and feeds-stuff, sterigmatocystion is more harmful than aflatoxins. The reported detection methods of sterigmatocystion included the Thin-layer Chromatography, the High-Performance-Liquid Chromatography, the Enzyme-Linked Immunosorbant Assay and the PCR detection to the toxic gene, however studies about both easy and inexpensive electro-chemical methods have not been found. Our previous studies had discovered that Sterigmatocystin (ST) exist similar sensitivity towards aflatoxin-detoxifizyme (ADTZ), which we had isolated from a fungus, as aflatoxin does. In this work, the preliminary study on electrochemical analysis and determination of ST with triplet electrode enzyme-biosensor system (Ag/AgCl as the reference electrode, Pt and Au as the pair and work electrode, respectively) was carried out. Multiwall-carbon-nanotube (MWNT) had been used to increase the electron transportation on electrode. In the research, the Au electrode was modified by MWNT-immobilized ADTZ, and then the voltammertric behavior of ST was studied by means of cyclic voltammogram analysis and different pulse analysis. Autoprobe CP Research Atomic Force Microscope and TECNAI 10 Transmission Electron Microscope, had been used to detect the MWNT as well as the surface of MWNT-modified ADTZ. The voltammertric behavior of ST was studied by means of cyclic voltammogram analysis and different pulse analysis. The results show that the red-ox peak potential of ST is at the point of -600 mV, the linear detection range is from 8.32 x 10(-5) to 66.56 x 10(-5) mg/mL, the detection limit is at 8.32 x 10(-5) mg/mL, and the response time is 10 seconds. This study provided a good basic work for further research.
Biosensing Techniques ; methods ; Electrochemistry ; Microscopy, Atomic Force ; Microscopy, Electron, Transmission ; Nanotubes, Carbon ; chemistry ; Sterigmatocystin ; analysis

OBJECTIVETo provide practical method for microscopic authentication of traditional Chinese medicine Gusuibu and its adulterants.

METHODBy means of light microscope, scanning electron microscopy and tissue section techniques, the morphology, the size of the rhizome scales and their bearing position in the original plants of Gusuibu and its adulterants, i. e. Drynaria roosii, D. delavayi, D. quercifolia and Pseudodrynaria coronans were analyzed.

RESULTThere were significant differences between scales length of D. roosii, D. delavayi and P. coronans, while there was no significant difference between that of D. roosii and D. quercifolia. The scale teeth of D. delavayi were usually curved, bifid and uneven distributed at the scale fringe, which was different from that of the other three species. The base of the scales sinks in epidermis in D. roosii, D. quercifolia, and P. coronans, while it bore at the raised part of epidermis in D. delavayi.

CONCLUSION[corrected] Morphology, size and bearing position of the rhizome scales have significant differences in the several species. Therefore, these characteristics can be applied to the identification of Gusuibu and its adulterants.

Medicine, Chinese Traditional ; methods ; Microscopy ; Microscopy, Electron, Scanning ; Polypodiaceae ; anatomy & histology ; classification ; Rhizome ; anatomy & histology ; classification