X-ray microanalysis--a method for determination of elements in bladder biopsies.

Chemical elements in biopsies of normal human urothelium and transitional cell carcinoma (TCC) are determined by X-ray microanalysis in combination with scanning electron microscopy. The methodology presented is intended to be used in subsequent studies to compare data of normal urothelium and TCC of different stage and grade.

Particulate matter contamination in the corneal stroma of severe eye burns in humans.

Corneal buttons obtained from keratoplasty were examined by energy dispersive x-ray analysis (EDXA) combined with scanning electron microscopy (SEM). This method enables to assay the mineral composition of minute parts of tissue samples identified in SEM images. Samples were cut from paraffin embedded corneae, deparaffinized in xylol, dried in aceton, critical-point desiccated, covered by evaporating with a thin layer of carbon and examined by SEM. In healthy human donor eyes, only some iron particles had been found. In the 22 patients samples high amounts of different particles were identified, materials from rubber stoppers, chromesteel, titanium pigments, talcum, barium and glass. Furthermore a lot of different metal particles containing varying amounts of Na, Mg, Al, Si, P, S, Cl, K, Ca, Fe, Cu, Cr, Zn, La and Ce were detected. Some particles may be caused by the initial trauma, others by therapy. Such contaminations might have supported leucocyte and fibrocyte invasion increasing the inflammatory reaction in the burnt cornea.

Measurement of BBN-induced alterations in rat urothelium by electron microscopic X-ray microanalysis.

The aim of this study was to analyse N-butyl-n-butanol-4-nitrosamine (BBN)-induced alterations of the urothelium in rats concerning its content of phosphorus, sulphur, chlorine, potassium and calcium using electron microscopic X-ray microanalysis (REM analysis). The following histopathological findings of the bladder mucosa were discovered after exposure to BBN: normal urothelium (n = 36); focal epithelial proliferations (n = 12) following 6-12 week's exposure; epithelial hyperplasia (n = 8) after urothelial carcinoma (n = 4) following 12 weeks' exposure. The observed phosphorus/sulphur and phosphorus/calcium ratios based on REM analysis did not show any statistical correlation with the morphological changes classified by light microscopy. Our data do not support the hypothesis raised by other investigators that an increase in phosphorus content or phosphorus/sulphur or phosphorus/calcium ratio could indicate early neoplastic transformations of urothelial cells as "tumor markers".

A simple empirical calibration of energy dispersive X-ray analysis (EDXA) on the cornea.

Monitoring of the corneal electrolyte content is important for the study of chemical eye burns. This paper describes quantitative measurements on gelatin standards, corneas and a cornea homogenate with an energy dispersive X-ray analyzer (EDX) in the scanning electron microscope (SEM). Ten micrometers thick cryosections were freeze-dried and mounted on solid carbon supports. The applied quantification procedure was a local peak background analysis with a specifically designed computer program. Similar chemical and physical properties of gelatin, cornea homogenate, and cornea were proven by EDX-analysis and wet chemical analysis. Gelatin standards with known concentrations of different added salts showed linear correlations with a correlation coefficient higher than 0.95 for all considered elements. The local background generation on carbon supports was the same for gelatin standards and corneal tissue. The results demonstrate that quantitative EDX analysis of semi-thin samples, mounted on neutral carbon supports, can be reliably used for the assessment of the corneal mineral composition.

Conjunctival tissue examination in severe eye burns: a study with scanning electron microscopy and energy-dispersive X-ray analysis.

BACKGROUND: Long-lasting inflammation is a major problem in treatment after severe eye burns and may find expression in an altered elemental composition of the conjunctiva. Particulate contamination of biological tissue induces such inflammatory processes. In the anterior eye segment, trauma or subsequent therapy may give rise to such contamination. Scanning electron microscopy and energy-dispersive X-ray analysis are able to detect traumatic residues of submicron size and changes of the elemental composition. METHODS: Conjunctival specimens from first-time peridectomy of three healthy and nine severely burnt-eyes were examined with scanning electron microscopy and energy-dispersive X-ray analysis. The samples were prepared as cryo- or paraffin sections, mounted on carbon blocks and coated with evaporated elemental carbon. RESULTS: The samples of healthy conjunctiva showed higher concentrations of Na, P and CI. These elements showed lower concentrations in conjunctival stroma of burnt eyes excised before the 20th day after trauma than in material obtained subsequently. In two burnt conjunctival specimens there was severe traumatic contamination with Ca in Ca(OH)2 and CaO burns, and in one case the traumatic substance was Si, in a peroxide plus silicone spray burn. In the remaining six cases, particulate contamination with Fe, Al, Ni, Zn, Cu, Ti and other substances was present in the burnt conjunctivas, while no contamination was detected in the specimens of healthy conjunctivas. CONCLUSIONS: The origin of the contaminant particles is assumed to be the trauma itself and the subsequent therapy. These investigations stress the importance, for clinical purposes, or early peridectomy and contamination-free therapy.

Scanning electron microscopic and energy-dispersive X-ray analysis findings on two brand new intraocular lenses.

Two intraocular lenses (IOLs) from the same manufacturer showed surface particles by scanning electron microscopy. With energy-dispersive X-ray analysis, these were shown to contain Mg-Si, Al-Mg-Si, Ti, Fe, Ca and Cu. A sterile chronic intraocular inflammation may result from implantation of such contaminated IOLs.

Does the human cornea contain silicon?

BACKGROUND: Our study investigated the presence, type and quantity of silicon in the human cornea. We report the results of silicon measurements in the corneas of silicotic individuals, bricklayers and apparently normal human individuals and offer a hypothesis for the mechanism of silicon deposition in the human cornea. METHODS: We examined corneas from 13 decreased subjects who suffered from silicosis, 2 bricklayers and 6 apparently healthy subjects. Cornea samples were examined by energy-dispersive x-ray analysis (EDXA) under calibrated conditions in a scanning electron microscope (SEM). The EDXA detector was a silicon-free germanium crystal. Five distinct layers (epithelium, Bowman's membrane, central stroma. Descemet's membrane and endothelium) were analyzed in each cornea. The method allows simultaneous semiquantitative analysis of, among other elements, silicon, calcium and oxygen. We measured amorphous silicon and visible particles of silicon. RESULTS: We found amorphous silicon in low concentrations in 38% of the silicotic corneas and in very low concentrations in 29% of the healthy corneas. Bricklayers showed high concentrations of amorphous silicon. These accumulations of silicon were predominantly located in Descemet's membrane. Silicotic corneas showed significantly more silicon-containing particles than corneas of healthy controls (chi 2-test, P < 0.01). CONCLUSION: Normal corneas contain very low amounts of silicon. Longterm exposure to inhalative silicon dusts results in only very slightly increased levels of amorphous silicon in the cornea. However, silicon-containing particles accumulate in the cornea of silicotic individuals. Bricklayers incorporate more amorphous silicon into the cornea.

Changes in mineral composition of rabbit corneas after alkali burn.

BACKGROUND: Mineral balance is essential for maintaining corneal transparency. In preliminary investigations, we found alterations of mineral concentrations in the human cornea after chemical burns. In this prospective study the mineral content of the rabbit cornea was quantified after experimental alkali burns. METHODS: Thirty-two eyes of rabbits were burnt for 30 s with 1 M NaOH. A short period of saline rinsing followed, but no further therapy was given. The eyes of 20 healthy rabbits served as a control. We measured the water content by weighing the cornea before and after drying. The mineral content of the cornea was determined in different stromal layers using energy-dispersive X-ray analysis in the scanning electron microscope. Groups of eight rabbits were examined on days 0, 1, 4 and 8 after the experimental burns. RESULTS: After alkali burns the mineral composition of the corneal stroma changed considerably. Sodium was present in high concentrations immediately after burning, up to 708 mmol/kg dry weight, and returned to normal on day eight. Chlorine increased significantly throughout the whole period. Sulphur concentrations stayed almost normal, showing only a minute loss of ground substance after alkali burn. A considerable and increasing edema could be concluded from the water content. CONCLUSIONS: Alkali burns resulted in a dramatic change of the corneal mineral content. Therapeutic intervention with appropriate solutions is required to restore and maintain the normal mineral composition of the denuded corneal stroma.

[Particulate contamination of infusion solutions and drug additives within the scope of long-term intensive therapy. 1. Energy dispersion electron images in the scanning electron microscope-REM/EDX]. / Partikuläre Verunreinigung von Infusionlösungen und Medikamentenzusätezen. im Rahmen einer Langzeit-Intensiv-Therapie. Teil 1. Rückstreuelektronenbilder im Rasterelektronenmikroskop-REM/EDX.

During use of i.v. solutions, particulate matter may be introduced into the patient. X-ray analyses have shown that the particulate matter consists mainly of glass from ampules, rubber from stoppers of infusion bottles, and plastic from infusion sets. A new method is introduced: scanning electron microscopy-SEM- in combination with energy-dispersive X-ray analysis-EDX. using this technique all foreign particles larger than 0.8 microns can be detected and analyzed in any tissue. Especially in critically ill patients, particularly matter is deposited in the microcirculation of the lung. The particles cause a variety of pathologic changes (formation of thrombi, thromboembolization of the microcirculation, destruction of the vascular endothelium, formation of granulomas and foreign body giant cells). The adult respiratory distress syndrome could be produced or respiratory insufficiency might be aggravated following circulatory shock. It is therefore recommended that terminal in-line filters with small pore sizes be used for all i.v. solutions.

[The filter effectiveness of common 15-micron filters (DIN 58362). II: Scanning electron microscopy and roentgen analysis]. / Untersuchung über die Filtereffektivität herkömmlicher 15-microns-Filter (DIN 58362). Teil II: Rasterelektronenmikroskopie und Röntgenanalyse.

When i.v. solutions are used, millions of foreign particles are washed into the organism. Our town studies could recently show that these particles may interact with the organism and thus cause injury. A new method is introduced here (energy dispersive X-ray analysis in scanning electron microscopy, SEM, Jeol 35 CF/EDX, EEDS II Ortec), with which the element combination of foreign particles can be analyzed qualitatively and quantitatively. As a first step an analysis of the particulate matter in filtrates of infusion solutions was carried out in the same way, materials used in infusion therapy (e.g. infusion sets, cannulas, etc.) were analyzed. The results show that most of the particulate matter consists of glass from ampoules, plastic materials from infusion sets and rubber from the rubber stoppers of infusion bottles.

[Filtering effectiveness of conventional 15 m micron filters (DIN 58362) and newly developed 15 m micron and 5 m micron filters. I: Particle count]. / Untersuchung über die Filtereffektivität herkömmlicher 15-microns m-Filter (DIN 58362) und endständiger 15-microns m-und 5-microns m-Filter. Teil I: Partiklzählung.

When i.v. solutions are used, a considerable amount of particulate matter is washed into the patient. This particulate matter consists mainly of glass from ampoules, plastic materials from infusion sets, rubber from the rubber stoppers of infusion bottles and various materials, partly derived from the manufacturing process of the solutions. A new method of particle counting is introduced here (modified thrombocyte counter, Becton Dickinson), whereby particulate matter (less than 2 microns, greater 0.8 microns) could be counted for the first time. Using this counting method, the efficiency of the standard filtration (DIN 58362) was tested. Also tested were 2 new final filtration systems (with 15 microns and 5 microns pore-size). All 3 filtration methods were shown to be ineffective; especially small particles (less than 2 microns) cannot be eliminated by these filtration methods.

[Particulate contamination of infusion solutions and drug additives in the framework of long-term intensive therapy. 2. An animal model]. / Partikuläre Verunreinigung von Infusionslösungen und Medikamentenzusätzen im Rahmen einer Langzeit-Intensiv-Therapie. Teil 2. Tiermodell.

When IV solutions are used, particulate matter may be introduced into the patient. Using X-ray studies we could demonstrate particulate matter consisting mainly of glass from ampules, rubber from rubber stoppers of infusion bottles, and plastic material from IV sets. With a recently introduced method of back-scatter pictures in scanning electron microscopy combined with X-ray studies--Jeol 35 CF/EEDS II Ortec (REM EDX)--we analyzed this phenomenon using an animal model (rabbit). It appears that particulate matter is deposited in the interstitial lung tissue and especially the pulmonary microcirculation. Pathogenetic effects could also be demonstrated (formation of thrombi, etc.).

[Silver nitrate burn after Credé's preventive treatment. A roentgen analytic and scanning electron microscopy study]. / Silbernitratverätzung nach Credéscher Prophylaxe. Eine röntgenanalytische und rasterelektronenmikroskopische Untersuchung.

Following Credé's prophylaxis with silver-nitrate, the cornea of a newborn presented greyish-brown, lime-like plaques on the nasal part of the right eye. A paracentral ulcerating stromal opacification undermined these appositions, when the patient was admitted to the eye-clinic at Aachen. In the material taken in a lamellar keratectomy scanning electron microscopical examination was able to prove the existence of granules, previously described in light-microscopy. These granules measured 100 to 300 nm in diameter and were placed up to 110 microns deep into the corneal stroma of the specimen. An earlier chemical analysis of necrotic material showed no silver specific reaction. By means of EDX-Analysis these granules could be identified as silver-containing. This was once reassured by a newly developed modification of van-Kossa's-staining-method. The fact that the granular deposits contained mainly silver proves that the onset of a sodium-chloride-irrigation did not promote an intended therapeutic silver-chloride-precipitation and therefore had no effects on the silver-nitrate's penetration abilities. Injuries by silver-nitrate-solutions used for Credé's prophylaxis are seldom but still reported. The mechanism of injury in this case of a child, born by sectio remains unknown. Neither the use of an unusual silvernitrate solution, that was taken from a disposable ampoule (Mova-Nitrat) was reported, nor any corneal injury during sectio mentioned.(ABSTRACT TRUNCATED AT 250 WORDS)

Corneal silver deposits following Crede's prophylaxis an examination with electron dispersive x-ray analysis (EDX-analysis) and scanning electron microscope (SEM).

In a case of silver nitrate injury after Credé's prophylaxis, the cornea of a newborn presented yellow-brown, lime-like plaques on the nasal part of the right eye. A paracentral ulcerating stromal opacification undermined these appositions, when the patient was admitted to the eye-clinic at Aachen. In the material obtained by a lamellar keratectomy scanning electron microscopical examination was able to prove the existence of granules, previously described in light-microscopy. These granules measured 100 to 300 nm in diameter. An earlier chemical analysis of necrotic material showed no silver specific reaction. By means of EDX-analysis these granules could be identified as silver-containing. Injuries by silver nitrate solutions used for Credé's prophylaxis are seldom but still reported. The mechanism of injury in this case of a child, born by sectio remains unknown. Neither the use of an unusual silver nitrate solution, that was taken from a disposable ampoule (Mova-Nitrat R) was reported, nor any corneal injury during sectio mentioned. Nevertheless the method of EDX-analysis and SEM proved the diagnosis of corneal silver deposits, so that the origin of the granullar opacification of the cornea could be determined.

Impact of particulate contamination in crystalloid cardioplegic solutions: studies by scanning and transmission electron microscopy.

The amount of particulate matter present in Bretschneider's cardioplegic solution (HTK) was assessed by laser-mediated particle counting. Permissible levels of contaminant particles with a distribution of diameters between 0.2 and 20 microns were found. A significant further increase in the particle count was observed when the fluid was administered for clinical use, which resulted in the additional release of particles from, for example, the infusion kit, which included an in-line filter with pores of 270 microns. Filtration of the HTK solution by a terminal inline filter (0.2 micron) significantly reduced the number of particles. In order to determine the chemical composition and the potential hazards of the particulate material we used scanning electron microscopy in combination with energy dispersive X-ray analysis and transmission electron microscopy to examine specimens taken from heart tissue obtained from Göttinger minipigs after cardioplegia and from humans undergoing mitral valve replacement after cardioplegia and reperfusion. Particles of various diameters were found either to be plugging coronary capillaries, to be adherent to the endothelial layer, or to be engulfed by polymorphonuclear (PMN) granulocytes, which appeared to be activated. Some of the PMN granulocytes were apparent in the endothelial layer. It is recommended, therefore, that a terminal in-line filter (0.2 micron) should be routinely used.