Polarization microscopy, possibly together with some contrast techniques (dark field and color phase contrast), was used to study the periphyton (microbiome) growing on filamentous green algae. The material containing filamentous algae with periphyton on the surface was collected in the villages of Sýkořice and Zbečno (Křivoklátsko Protected Landscape Area). The objects were studied in a LOMO MIN-8 St. Petersburg polarizing microscope and a Carl Zeiss Jena NfpK laboratory microscope equipped with an In Ph 160 basic body with variable dark field or color phase contrast and a Nikon D70 DSLR digital camera. Cells of filamentous algae of the genera Cladophora, Vaucheria, and Oedogonium were studied and the periphyton attached to them formed by cyanobacteria of the genera Chamaesiphon and Pleurocapsa and algae of the genera Characium, including diatoms of the genera Eunotia and Synedra. In all cases, the cell walls of the host algae showed a very strong birefringence. In contrast, the walls of cyanobacteria of the genera Chamaesiphon and Pleurocapsa were characterized by a much weaker birefringence (Pleurocapsa somewhat thicker), and the diatom frustules of the genera Eunotia and Synedra were almost without a birefringence. Strongly birefringent granules were found in the cytoplasm of the green alga of the genus Characium, which forms periphyton on the filamentous green algae of the genus Vaucheria. The periphyton on the filamentous alga of the genus Oedogonium, formed by cyanobacteria of the genus Pleurocapsa and diatoms of the genera Eunotia and Synedra, deposited in a massive layer of mucus containing birefringent crystals, showed a particularly strong birefringence. At the end of the vegetation of filamentous algae, their parts and remnants of periphyton (diatom frustules and crystals) became part of the detritus at the bottom of the culture vessel. The use of polarization microscopy in the study of filamentous algae with periphyton on the surface allows us not only to determine the birefringence of the observed structures, but also to partially deduce their chemical composition, or regular arrangement of particles, so-called shape birefringence.
Polarization and positive phase contrast microscope were concomitantly used in the study of the internal structure of microbial cells. Positive phase contrast allowed us to view even the fine cell structure with a refractive index approaching that of the surrounding environment, e.g., the cytoplasm, and transferred the invisible phase image to a visible amplitude image. With polarization microscopy, crossed polarizing filters together with compensators and a rotary stage showed the birefringence of different cell structures. Material containing algae was collected in ponds in Sýkořice and Zbečno villages (Křivoklát region). The objects were studied in laboratory microscopes LOMO MIN-8 Sankt Petersburg and Polmi A Carl Zeiss Jena fitted with special optics for positive phase contrast, polarizers, analyzers, compensators, rotary stages, and digital SLR camera Nikon D 70 for image capture. Anisotropic granules were found in the cells of flagellates of the order Euglenales, in green algae of the orders Chlorococcales and Chlorellales, and in desmid algae of the order Desmidiales. The cell walls of filamentous algae of the orders Zygnematales and Ulotrichales were found to exhibit significant birefringence; in addition, relatively small amounts of small granules were found in the cytoplasm. A typical shape-related birefringence of the cylindrical walls and the septa between the cells differed in intensity, which was especially apparent when using a Zeiss compensator RI-c during its successive double setting. In conclusion, the anisotropic granules found in the investigated algae mostly showed strong birefringence and varied in number, size, and location of the cells. Representatives of the order Chlorococcales contained the highest number of granules per cell, and the size of these granules was almost double than that of the other monitored microorganisms. Very strong birefringence was exhibited by cell walls of filamentous algae; it differed in the intensity between the cylindrical peripheral wall and the partitions between the cells. Positive phase contrast enabled us to study the morphological relationship of various fine structures in the cell (poorly visible in conventional microscope) to anisotropic structures that have been well defined by polarization microscopy.
The pathophysiology of microcirculation is intensively investigated to understand disease development at the microscopic level. Orthogonal polarization spectral (OPS) imaging and its successor sidestream dark-field (SDF) imaging are relatively new noninvasive optical techniques allowing direct visualization of microcirculation in both clinical and experimental studies. The goal of this experimental study was to describe basic microcirculatory parameters of skeletal muscle and ileal serous surface microcirculation in the rat using SDF imaging and to standardize the technical aspects of the protocol. Interindividual variability in functional capillary density (FCD) and small vessels (<25 microm in diameter) proportion was determined in anesthetized rats on the surface of quadriceps femoris (m. rectus femoris and m. vastus medialis) and serous surface of ileum. Special custom made flexible arm was used to fix the SDF probe minimizing the pressure movement artifacts. Clear high contrast images were analyzed off-line. The mean FCD obtained from the surface of skeletal muscle and ileal serous surface was 219 (213-225 cm/cm(2)) and 290 (282-298 cm/cm(2)) respectively. There was no statistically significant difference between rats in mean values of FCD obtained from the muscle (P = 0.273) in contrast to ileal serous surface, where such difference was statistically significant (P = 0.036). No statistically significant differences in small vessels percentage was detected on either the muscle surface (P = 0.739) or on ileal serous surface (P = 0.659). Our study has shown that interindividual variability of basic microcirculatory parameters in rat skeletal muscle and ileum is acceptable when using SDF imaging technique according to a highly standardized protocol and with appropriate fixation device. SDF imaging represents promising technology for experimental and clinical studies.
- MeSH
- Quadriceps Muscle blood supply MeSH
- Financing, Organized MeSH
- Ileum blood supply MeSH
- Rats MeSH
- Microcirculation MeSH
- Microscopy, Polarization methods standards MeSH
- Rats, Wistar MeSH
- Reproducibility of Results MeSH
- Serous Membrane blood supply MeSH
- Animals MeSH
- Check Tag
- Rats MeSH
- Male MeSH
- Animals MeSH
Spatial light modulators have become an essential tool for advanced microscopy, enabling breakthroughs in 3D, phase, and super-resolution imaging. However, continuous spatial-light modulation that is capable of capturing sub-millisecond microscopic motion without diffraction artifacts and polarization dependence is challenging. Here we present a photothermal spatial light modulator (PT-SLM) enabling fast phase imaging for nanoscopic 3D reconstruction. The PT-SLM can generate a step-like wavefront change, free of diffraction artifacts, with a high transmittance and a modulation efficiency independent of light polarization. We achieve a phase-shift > π and a response time as short as 70 µs with a theoretical limit in the sub microsecond range. We used the PT-SLM to perform quantitative phase imaging of sub-diffractional species to decipher the 3D nanoscopic displacement of microtubules and study the trajectory of a diffusive microtubule-associated protein, providing insights into the mechanism of protein navigation through a complex microtubule network.
- MeSH
- Time Factors MeSH
- Microscopy, Interference methods statistics & numerical data MeSH
- Metal Nanoparticles ultrastructure MeSH
- Humans MeSH
- Microscopy, Atomic Force MeSH
- Microscopy, Phase-Contrast methods statistics & numerical data MeSH
- Microtubules metabolism ultrastructure MeSH
- Nanotechnology MeSH
- Nanotubes ultrastructure MeSH
- Optical Phenomena MeSH
- Computer Simulation MeSH
- Microtubule-Associated Proteins metabolism MeSH
- Cell Cycle Proteins metabolism MeSH
- Schizosaccharomyces pombe Proteins metabolism MeSH
- Light MeSH
- Tubulin metabolism MeSH
- Gold MeSH
- Imaging, Three-Dimensional methods statistics & numerical data MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
Little is known about the viability of myxozoan actinospore stages after harvest from laboratory cultures of infected oligochaete worms. The viability and reactivity of actinospores of three myxozoan species was evaluated after short-term storage at 4°C and 12°C. Two methods of determining actinospore viability were compared: differential fluorescent staining and direct microscopic observation of morphological indicators of spore integrity. Spore reactivity was quantified by measuring polar filament discharge rates in a micro-assay with fish mucus substrate and mechanical stimulation by vibration. The age-dependent viability of the three species showed clear differences. Myxobolus cerebralis actinospores had the shortest effective life span whereas Henneguya nuesslini actinospores survived significantly longer. Storage at lower temperatures yielded higher viability in all species. Myxobolus pseudodispar actinospores were significantly robust up to 12°C when assessed by staining, but showed similar viability characteristics as H. nuesslini when analyzed morphologically. Evaluation of spore viability by fluorescent staining correlated with morphological assessment, although fewer viable actinospores were usually detected microscopically. Polar filament discharge activity of morphologically intact actinospores did not significantly decrease until the third day of storage compared to freshly harvested samples. The results indicate that durability and reactivity trends during storage of actinospores differ among myxozoan species.
Smulevich -- Optical and Infrared Absoфtion as a Probe of the Dynamics of Heme Proteins 6 -- S. S. Raman Microscope with a Scanning Electron Microscope 23 -- Y. y. Aksenov. E. G. J. Reinders, J. Fastermann, 1 Heberle -- A New Method Coupling Polarized ATR-FTIR Spectroscopy and iH/^H Exchange to Pankowska -- Structural and Optical Properties of LH2-CompIexes -- R. Kassies. S. Bahatyrova, C. N. Peretti -- Optical Interference Study of Physiological Fluids Transportation Through 109 -- Semipermeable
2 sv. (150 s., s. 151-274) : il., tab. ; 25 cm
- MeSH
- Molecular Biology MeSH
- Spectrum Analysis MeSH
- Publication type
- Abstracts MeSH
The aim of this study was to evaluate the efficiency of using meiotic spindle (MS) visibility and relative position to the polar body (PB) as indicators of oocyte maturation in order to optimize intracytoplasmic sperm injection (ICSI) timing. This was a cohort study of patients younger than 40 years with planned ICSI, the timing of which was determined by MS status, compared with those without MS evaluation. The angle between PB and MS and MS visibility were evaluated by optical microscope with polarizing filter. Oocytes with MS evaluation were fertilized according to MS status either 5-6 h after ovum pick-up (OPU) or 7-8 h after OPU. Oocytes without MS evaluation were all fertilized 5-6 h after OPU. For patients over 35 years visualization of MS influenced pregnancy rate (PR): 182 patients with MS visualization had 32% PR (58/182); while 195 patients without MS visualization had 24% PR (47/195). For patients under 35 years, visualization of MS did not influence PR: 140 patients with MS visualization had 41% PR (58/140), while 162 patients without MS visualization had 41% PR (66/162). Visualization of MS therefore appears to be a useful parameter for assessment of oocyte maturity and ICSI timing for patients older than 35.
Cieľ práce: opis a imunohistochemická analýza prednádorových zmien a nádora obličky u pacienta s chronickou obličkovou nedostatočnosťou liečenou dlhodobou hemodialýzou. Metódy: Nefrektomický bioptický materiál (ľavá oblička), bol vyšetrený mikroskopicky a polaroskopicky. Ďalej boli na analýzu použité tieto protilátky: CK1/CK3, CK5/6, CK8, CK20, EMA, „Renal cells“, CD10, Ki-67, PCNA, p53 a E-cadherin. Hlavné nálezy: Operačný preparát pochádzal z pacienta, liečeného pre renálnu insuficienciu 11 rokov hemodialýzou pre diabetickú nefropátiu a chronickú tubulointersticiálnu nefritídu. Indikáciou pre urgentnú nefrektómiu bola bolesť a masívne intrarenálne krvácanie. Spolu s nálezom vyššie uvedených obličkových ochorení zistili sme rozvinutý nález získanej cystózy vznikajúcej počas dlhodobej dialýzy, viaceré malé papilárne adenómy a multifokálne karcinómy z buniek obličky vykazujúce konvenčný, ďalej papilárny a sarkomatoidný charakter. Prítomné boli tiež známky ťažkej vaskulárnej nefrosklerózy a uremickej oxalózy. Horný pól orgánu bol masívne prekrvácaný. Závery: Nami popísaný nález obsahuje súčasnú prítomnosť mikroskopických zmien vysvetľujúcich stav chronickej obličkovej nedostatočnosti s prítomnou uremickou oxalózou, kombinovaných so získanou cystózou vznikajúcou v dôsledku dlhodobej hemodialýzy. V takto zmenenom orgáne sa nachádzali dysplastické mikrotubulárne a intracystické zmeny, smerujúce ku vzniku viacerých papilárnych adenómov a multifokálnych karcinómov z buniek obličky, vyznačujúcich sa rôznorodou diferenciáciou.
Purpose of the investigation: Description of precancerous lesions and kidney tumors developing in a patient with chronic uremia treated by long-term hemodialysis. Most important methods. Light microscopy, polarization and immunohistochemistry with CK1/CK3, CK5/6, CK7, CK8, CK20, EMA, Renal cell, CD10, Ki-67, PCNA, p53 and E-cadherin antibodies were used. Main findings: After 11 years of hemodialysis treatment of end-stage diabetic nephropathy and chronic tubulointerstitial nephritis an urgent left-sided nephrectomy was performed because of pain and massive intrarenal bleeding. Biopsy revealed acquired cystic kidney disease associated with multiple precancerous lesions, several small papillary adenomas and a multifocal renal cell carcinoma with conventional and papillary structures with admixture of small foci of highly cellular sarcomatoid features. Severe vascular nephrosclerosis and uremic oxalosis were additional findings. The upper pole of the kidney was massively hemorrhagic. Principal conclusions: This case illustrates the association of chronic renal insufficiency, uremic oxalosis, long-term hemodialysis, acquired cystic kidney disease and development of variable precursor intratubular and intracystic lesions progressing to several papillary adenomas and multifocal renal cell carcinomas with variegated microscopic structures in one kidney.
- MeSH
- Biopsy MeSH
- Renal Insufficiency, Chronic diagnosis therapy MeSH
- Kidney Diseases, Cystic diagnosis etiology therapy MeSH
- Renal Dialysis methods utilization MeSH
- Immunohistochemistry methods utilization MeSH
- Disease Attributes MeSH
- Pathology, Clinical methods trends MeSH
- Humans MeSH
- Microscopy MeSH
- Kidney Neoplasms diagnosis etiology therapy MeSH
- Nephrectomy MeSH
- Precancerous Conditions diagnosis etiology therapy MeSH
- Aged MeSH
- Check Tag
- Humans MeSH
- Male MeSH
- Aged MeSH
- Publication type
- Case Reports MeSH
Článek je pokračováním pojednání (I) o technice, resp. bioinženýrských metodách pro praxia výzkum v dermatovenerologii. V tomto II. oddíle jsou probrány vyšetřovací metody určené k zobrazováníkůže a jejích struktur. Lze je rozdělit na přímé a nepřímé. V první části přímých metod jevěnována pozornost klasické fotografii a nové digitální fotografii. Jsou porovnány vlastnosti, technickémožnosti a přednosti obou způsobů, kvalita a využití. Probrány jsou způsoby archivace,počítačového zpracování, komprimace dat.Zmíněny jsou i způsoby makro- a mikrofotografií, včetně stereomikroskopie. Mezi nové metodypatří i videomikroskopie a její provádění v polarizovaném světle.Tato elektronická mikroskopie poskytuje při spojení s příslušným softwarem možnost ještěkvalitnějšího zpracování dat, především kožních nádorů (melanomů) a névů. Počítačové posuzovánína principu tzv. neuronových sítí poskytuje ještě kvalitnější ABCD+E+T analýzu s vyšší pravděpodobnostízáchytu melanomu. Systém je využitelný i pro ostatní uchování a popis obrazů. Přípojtlačítkem„video-mail“ dává možnost použití v teledermatologii, tj. při teledermatoskopii, teledermatopatologii,při konzultacích a edukaci.V další části jsou uvedeny mikroskopické metody, jako jsou konfokální a elektronová (skenovací)mikroskopie s jejich konstrukčními a technickými parametry a využitím.Do stati přímých metod patří ještě kapilaroskopie a fluorescenční videomikroskopie, optickákoherentní tomografie kůže a protonová magnetická rezonance kůže.K nepřímým metodám zařazujeme dopplerometrii (A-, B-, C-, M-mode scan), laser-Doppler-průtokometrii,měření nerovnosti kožního povrchu (profilometrii) a kožní viziometr. Jsou opět uvedenyjejich konstrukční základy, s technickým a praktickým použitím.
The article is a continuation of part I dealing with the technique and bioengineering methods forpractice and research in dermatovenerology. In part II the authors discuss examination methods forimaging of the skin and its structures. They can be divided into direct and indirect ones. In the firstpart of direct methods attention is paid to classical photography and new digital photography. Theauthors compare properties, technical possibilities and advantages of both methods, their qualityand application. They discuss also methods of archivation, computer processing and comprimationof data.The authors mention methods of macro- and microphotography, incl. stereomicroscopy. Newmethods include also videomicroscopy incl. its implementation in polarized light.This electronic microscopy provides when combined with the appropriate software the possibilityof high standard processing of data, in particular skin tumours (melanomas) and naevi. Computer-assisted evaluation on the principle of so-called neuron networks makes even higher-standardABCD+E+T analysis possible with a greater probability of melanoma detection. The system can beused also for other preservation and description of images. Connection by button „video-mail“ makesit possible to use of in teledermatology, i.e. in teledermatoscopy, teledermatopathology, duringconsultations and education. In the subsequent part microscopic methods are presented such as confocal and electron(scanning) microscopy with their design, technical parameters and use.The section on direct methods includes also capillaroscopy and fluorescent videomicroscopy,optic coherent tomography of the skin and proton magnetic resonance of the skin.Indirect methods include dopplerometry (A-, B-, C-, M-mode scan), laser-doppler flowmetry,assessment of inequalities of the skin surface (profilometry) and skin visiometry. Their design,technical and practical use are described.
- MeSH
- Dermatology methods MeSH
- Microscopy, Electron methods utilization MeSH
- Microscopy, Confocal methods utilization MeSH
- Humans MeSH
- Magnetic Resonance Imaging methods utilization MeSH
- Ultrasonography, Doppler methods utilization MeSH
- Microscopy, Video methods utilization MeSH
- Check Tag
- Humans MeSH
- Publication type
- Review MeSH
