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Biomedical subjects

V Sarafis

Publications and source records attributed to V Sarafis.

12 recordsLinked to original sources

Identification of Photosystem I and Photosystem II enriched regions of thylakoid membrane by optical microimaging of cryo-fluorescence emission spectra and of variable fluorescence.

Oxygenic photosynthesis of higher plants requires linear electron transport that is driven by serially operating Photosystem II and Photosystem I reaction centers. It is widely accepted that distribution of these two types of reaction centers in the thylakoid membrane is heterogeneous. Here, we describe two optical microscopic techniques that can be combined to reveal the heterogeneity. By imaging micro-spectroscopy at liquid nitrogen temperature, we resolved the heterogeneity of the chloroplast thylakoid membrane by distinct spectral signatures of fluorescence emitted by the two photosystems. With another microscope, we measured changes in the fluorescence emission yield that are induced by actinic light at room temperature. Fluorescence yield of Photosystem II reaction centers varies strongly with light-induced changes of its photochemical yield. Consequently, application of moderate background irradiance induces changes in the Photosystem II fluorescence yield whereas no such modulation occurs in Photosystem I. This contrasting feature was used to identify regions in thylakoid membranes that are enriched in active Photosystem II.

Image Processing, Computer-Assisted↗

Localization and movement of mineral oil in plants by fluorescence and confocal microscopy.

Fluorescence and confocal laser scanning microscopy were explored to investigate the movement and localization of mineral oils in citrus. In a laboratory experiment, fluorescence microscopy observation indicated that when a 'narrow' distillation fraction of an nC23 horticultural mineral oil was applied to adaxial and opposing abaxial leaf surfaces of potted orange [Citrus x aurantium L. (Sapindales: Rutaceae)] trees, oil penetrated steadily into treated leaves and, subsequently, moved to untreated petioles of the leaves and adjacent untreated stems. In another experiment, confocal laser scanning microscopy was used to visualize the penetration into, and the subsequent cellular distribution of, an nC24 agricultural mineral oil in C. trifoliata L. seedlings. Oil droplets penetrated or diffused into plants via both stomata and the cuticle of leaves and stems, and then moved within intercellular spaces and into various cells including phloem and xylem. Oil accumulated in droplets in intercellular spaces and within cells near the cell membrane. Oil entered cells without visibly damaging membranes or causing cell death. In a field experiment with mature orange trees, droplets of an nC23 horticultural mineral oil were observed, by fluorescence microscopy, in phloem sieve elements in spring flush growth produced 4-5 months and 16-17 months after the trees were sprayed with oil. These results suggest that movement of mineral oil in plants is both apoplastic via intercellular spaces and symplastic via plasmodesmata. The putative pattern of the translocation of mineral oil in plants and its relevance to oil-induced chronic phytotoxicity are discussed.

Biological Transport↗

Improvement of optical resolution in far-field imaging by optical multiplication.

Based on the idea of the complex filters suggested by Toraldo di Francia in 1952, super-resolved imaging has been achieved by using interferometric image multiplication. The question of whether super-resolution implies a limited field of view is discussed, and an analytical criterion to decide this question is obtained. Three different imaging configurations are investigated, all of which gave a spatial resolution exceeding that corresponding to the full open aperture of the optical system. A resolution limit 55% of the Rayleigh limit was achieved for incoherently illuminated objects. One price that must be paid for the super-resolution is inefficient use of the light source. A second price is a limitation to the usable field of view.

Fourier Analysis↗

Optical sectioning by two-pinhole confocal fluorescence microscopy.

A two-pinhole axially superresolving confocal fluorescence imaging system is presented. Based on the concept of subtractive incoherent imaging, the system described here is equipped with a zero-focus complex-transmittance pupil filter in one of the collector paths. The optical sectioning capacity of the system is 25% superior to that of a free-pupil one-pinhole instrument.

Image Enhancement↗

Superresolution in far-field imaging.

We demonstrate the feasibility of achieving superresolved images by using Fourier-plane phase masks and image multiplication, which together create effective point-spread functions that are not positive definite and therefore cannot be created by any single Fourier plane mask in a linear system. Three different configurations were investigated, all of which gave a spatial resolution exceeding that corresponding to the full open aperture of the optical system. One price that must be paid for the superresolution is inefficient use of the light source.

Journal Article↗

The fluorescent dye 3, 3' dihexyloxacarbocyanine iodide selectively stains the midpiece and apical region of the heads of murid rodent spermatozoa.

Fluorescence microscopy of caudal epididymal spermatozoa stained with 3, 3' dihexyloxacarbocyanine iodide (DiOC6(3)) showed intense fluorescence along the concave surface of the apical hook of spermatozoa of Rattus species and along the upper concave margin of the sperm head in Mus musculus. In the spermatozoa of Hydromys chrysogaster, Melomys cervinipes, and Pseudomys australis, the two ventral processes also fluoresced brightly. In P. australis, fluorescence in the apical hook of sperm heads was largely localized to its upper and lower surfaces. The sperm of N. alexis did not show consistent positive fluorescence. The localization of fluorescence in these spermatozoa after staining with DiOC6(3) was mainly restricted to regions where a large accumulation of perinuclear theca material lies beneath the plasmalemma. The reason for this remains to be determined, but DiOC6(3) may be useful for quickly demonstrating areas of abundant perinuclear thecal material in sperm heads of eutherian mammals by light microscopy.

Animals↗

Applications of chemical-shift-selective NMR microscopy to the non-invasive histochemistry of plant materials.

Chemical-shift-selective imaging at microscopic resolution has been applied to the study of various plant materials including orange peel, grape berries and both dried and undried fruits of fennel. It is shown that selective imaging of aromatics and carbohydrates as well as water and oil can be performed with in-plane resolution down to 13 microns and slice thicknesses of 1 mm or less, at a field strength of 4.7 Tesla. The noninvasive nature of the method gives it advantages over established methods of plant histochemistry which involve sectioning and staining to reveal different chemical constituents.

Fruit↗

Localization of actin in the sperm head of the plains mouse, Pseudomys australis.

Actin was localized in the sperm head of the plains mouse, Pseudomys australis, using NBD-phallacidin, a fluorescent-labeled phallotoxin. It was found to be present in the two ventral hooks in sperm from the testis, epididymis, and vas deferens. Faint fluorescence was observed in other regions of the sperm head, whereas autofluorescence occurred in the midpiece of the tail in some preparations. The material in the ventral hooks was also found to be birefringent, which is consistent with it showing preferential orientation.

Actins↗

Sperm head morphology of the plains mouse Pseudomys australis.

Spermatozoa of plains mice were studied by transmission and scanning electron microscopy. The occurrence of a distinct dorsal hook and 2 ventral hooks joined at the base was confirmed. Acrosomal material covered the dorsal hook and appeared to constitute most of the 2 ventral hooks which contained nuclear material only at the base. Incubation in sodium dodecyl sulphate resulted in loss of all the material presumed to be acrosomal.

Animals↗