Confocal reader for biochip screening and fluorescence microscopy

We developed a fluorescence reader for the sensitive detection of surface-generated fluorescence. The system is applicable for high resolution imaging as well as for the readout of large biochips. The surface of a microscope coverslip is scanned with a laser beam focused to a waist diameter of 500 nm (FWHM) by means of a single aspheric lens. Scanning large areas with a focused beam usually evokes the need of automatic control elements to adjust the laser spot to the designated position at the surface. Due to the special design of the reader, the focus keeps at the plane of the surface even when scanning large areas, obviating the requirement of any real time control. Thus the instrument is straightforward and inexpensive. Nevertheless it features a high sensitivity and high optical resolution. The versatility of the instrument is demonstrated by imaging cells and reading out a DNA-chip. The excellent sensitivity is shown by detecting single fluorescently labeled antibodies.     

Changes in fibronectin staining in the human lens during ageing and cataractogenesis

The content and localization of fibronectin, an extracellular glycoprotein, in the serial sections of lenses of normal human donors and cataractous patients of different ages were determined by the indirect immunoperoxidase staining technique. This was followed by the evaluation with quantitative morphometric analysis. It was shown that fibronectin was present in the area of cell contacts as single deposits of faint orange-brown stained material in the lens samples of young donors. The fibronectin level was raised in lens sections from aged donors. Its accumulation was detected mostly within the spaces of the lens fiber cells. At different stages of cataractogenesis a dramatic decrease of the fibronectin content was detected in the lens sections obtained from patients of different ages. A new linear spectrophotometric technique was developed for evaluation of the lens transparency, to correlate the lens opacity with corresponding histological data obtained from the immunostaining technique. Morphological studies performed further suggested that the lens fiber cell plasma membrane structures were deteriorated. This was observed as changes of fibronectin staining in the lens sections at different periods of human ageing and cataract development. It is concluded that a decrease of fibronectin staining in the human lens is an indication for the structural damage of the lens fiber cell plasma membranes during ageing and cataractogenesis.     

Space-charge lens for focusing negative ion beams by means of an additional electron ionizer

A space-charge lens created at the Institute of Physics, National Academy of Sciences of Ukraine, to focus negative ion beams using an additional electron ionizer is investigated. In the previous version of the lens, in which the gas was ionized by the ion beam itself, the focal power was quite high (the focal length was ? ≤ 20 cm) but the gas pressure was too great (P ～ 10^?3 torr), which resulted in significant charge-exchange losses of the beam ions. The experimental and theoretical study reported here shows that the use of a 100-eV electron beam as an auxiliary ionizer allows the working pressure in the lens to be significantly reduced. As a result, a simple, inexpensive, and efficient lens has been developed that can be used in systems for transportation of negative ion beams.     

An optical multi-sensing system for detection of cardiovascular toxicity

A mini-microscope-based system for multisite detection of cardiovascular toxicity was developed. The mini-microscope consisted of an image sensor and lens module extracted from an inexpensive webcam. The flipped lens module enabled cells to be magnified and monitored during testing. The portability and compactness of this system enables short-term and potential long-term experimentation inside a conventional incubator. The toxicity test results demonstrated that the normalized beating rates of cardiac muscle cells selected from multiple regions increased over time when treated with 100 nM isoprenaline. The presented system could be a promising cost-effective cell-based testing tool for discovering and screening drugs.     

Preparation of leukocyte-poor erythrocyte concentrates using a simple aspiration technic

Depletion of leukocytes from red blood cells can prevent transfusion reactions in HLA-sensitized patients. We describe a simple technique of aspiration of the buffy-coat after centrifugation of red blood cells concentrates, less than 6 days old. This method can remove 87% of the leukocytes. The average leukocyte count after aspiration is 2 x 10(8). This method is inexpensive and does not require any special equipment.     

High-vacuum optical platform for cryo-CLEM (HOPE): A new solution for non-integrated multiscale correlative light and electron microscopy

Cryo-correlative light and electron microscopy (cryo-CLEM) offers a unique way to analyze the high-resolution structural information of cryo-vitrified specimen by cryo-electron microscopy (cryo-EM) with the guide of the search for unique events by cryo-fluorescence microscopy (cryo-FM). To achieve cryo-FM, a trade-off must be made between the temperature and performance of objective lens. The temperature of specimen should be kept below devitrification while the distance between the objective lens and specimen should be short enough for high resolution imaging. Although special objective lens was designed in many current cryo-FM approaches, the unavoided frosting and ice contamination are still affecting the efficiency of cryo-CLEM. In addition, the correlation accuracy between cryo-FM and cryo-EM would be reduced during the current specimen transfer procedure. Here, we report an improved cryo-CLEM technique (high-vacuum optical platform for cryo-CLEM, HOPE) based on a high-vacuum optical stage and a commercial cryo-EM holder. The HOPE stage comprises of a special adapter to suit the cryo-EM holder and a high-vacuum chamber with an anti-contamination system. It provides a clean and enduring environment for cryo specimen, while the normal dry objective lens in room temperature can be used via the optical windows. The ‘touch-free’ specimen transfer via cryo-EM holder allows least specimen deformation and thus maximizes the correlation accuracy between cryo-FM and cryo-EM. Besides, we developed a software to perform semi-automatic cryo-EM acquisition of the target region localized by cryo-FM. Our work provides a new solution for cryo-CLEM and can be adapted for different commercial fluorescence microscope and electron microscope.     

Radio gas chromatography of steroid metabolites by collection of radioactive fractions on thin layer chromatography plates

A simple and inexpensive technique for the monitoring of radioactive compounds in gas chromatographic effluent is described. Use has been made of the adsorption properties of thin layer chromatography plates to trap eluting radioactive peaks. Performance of the technique is demonstrated by analysis of radioactive steroid mixtures. The efficiency and sensitivity of this technique equal that of more complicated techniques.     

A method for concentrating dilute solutions of macromolecules

A simple, inexpensive acrylate polymer which has a capacity to absorb 170 ml of water per g has been developed. It can be used to concentrate dilute solutions of macromolecules such as proteins, nucleic acids, and carbohydrates. The polymer absorbs only low-molecular-weight substances such as glucose, sucrose, and inorganic salts. It can replace the various conventional concentration methods. No special device or electricity is needed for the concentration. The inexpensive polymer, molded in the form of rods, can be very conveniently used as “disposable concentration sticks.”     

Spectrofluorometric determination of bacterial DNA base composition

A spectrofluorometric technique for bacterial DNA base composition has been developed. This fast and simple technique requires two fluorescent dyes and a few inexpensive reagents. The data from this assay indicate that the guanine-cytosine content obtained was within acceptable statistical limits in comparison to commonly cited literature values. The spectrofluorometric technique is reliable and reproducible.     

Cross-linked leucaena seed gum matrix: an affinity chromatography tool for galactose-specific lectins

A cross-linked leucaena (Leucaena leucocephala) seed gum (CLLSG) matrix was prepared for the isolation of galactose-specific lectins by affinity chromatography. The matrix was evaluated for affinity with a known galactose-specific lectin from the seeds of snake gourd (Trichosanthes anguina). The matrix preparation was simple and inexpensive when compared to commercial galactose-specific matrices (i.e. about 1.5 US dollars/100 ml of matrix). The current method is also useful for the demonstration of the affinity chromatography technique in laboratories. Since leucaena seeds are abundant and inexpensive, and the matrix preparation is easy, CLLSG appears to be a promising tool for the separation of galactose-specific lectins.     

Vesicular and bullous diseases of the skin; cytologic diagnosis

A new technique, first described by Tzanck, for diagnosis of vesicular and bullous diseases of the skin by cytologic examination of material firmly scraped from lesions has a number of advantages over excision of biopsy specimens. It takes relatively little time, is painless, simple and inexpensive. The method also has value in research and teaching.     

VESICULAR AND BULLOUS DISEASES OF THE SKIN—Cytologic Diagnosis

A new technique, first described by Tzanck, for diagnosis of vesicular and bullous diseases of the skin by cytologic examination of material firmly scraped from lesions has a number of advantages over excision of biopsy specimens. It takes relatively little time, is painless, simple and inexpensive. The method also has value in research and teaching.     

A simplified lead oxide cadaver injection technique

A simple, inexpensive injectable substance is reported for analysis of the arterial circulation in fresh human or animal cadavers. The technique is a modification of that reported by Salmon in 1936 and utilizes lead oxide and gelatin. This combination is highly radiopaque; it perfuses the small radicles of the vascular tree and sets to a firm rubbery consistency to fascilitate dissection.     

复眼透镜光学信息保密编码及译码的研究——Ⅱ.分解掩模编码与综合再现

本文根据复眼透镜光学信息编译码的技术原理,实现了对二维图像进行分解编码记录以及综合译码再现.一幅m×n个目标单元的二维图像,通过1×k阵列的复眼透镜,得到(1×k)(m×n)个像元.经过一个特制的掩模板,得到一幅随机分解编码像,并根据透镜的物、像共轭原理,综合再现了原始图像.进而还实现了同时记录多幅二维图像信息的互补编码像,以及将互补编码像分离重现了每一幅原始目标图像.此互补编码像携带了更大的信息量,同时也大大提高了保密性能.     

A chronic intravenous cannulation technique adapted for behavioral studies

A simple inexpensive technique for chronic intravenous cannulation of small laboratory animals is described. With this technique multiple daily blood samples can be taken to form serial hormone profiles of individual animals. The procedure does not interfere with ongoing behavioral and physiological processes. As a measure of stress caused by the procedure, prolactin levels were determined by radioimmunoassay. Prolactin levels were only elevated on the first postoperative day; subsequent levels were comparable to those measured in decapitated animals. It is concluded that this technique can be very useful for short- and long-term experiments in the field of hormones and behavior.     

Centering mechanism for soft contact lenses

A soft contact lens, because of its elasticity, can re-center itself over the cornea after it has been displaced by a blink. In this paper, a potential energy method is used to simulate the centering mechanism. Specifications for commercially available soft contact lenses and realistic eye shapes are used for the examples. With this technique we can compare the centering attributes of different lens and eye geometries.     

Identification of Human Vital Functions Directly Relevant to the Respiratory System Based on the Cardiac and Acoustic Parameters and Random Forest

Regarding sleep research, polysomnography (PSG) also called a sleep study, is a gold standard. It incorporates brain waves, the oxygen level in the blood, heart rate and breathing, and leg movement recordings. PSG is a complicated and expensive laboratory-based procedure, usually done in hospitals or special sleep center. In this study, an alternative technique for Sleep-Related Breathing Disorders (SRBD) based on selected cardiac and acoustic parameters and the Random Forest (RF) has been studied. A system dedicated to the detection of simultaneously acquired ECG and acoustic signals, which are collected during sleep at home environment is proposed. Results obtained indicate that classification and regression tree models such as RF are appropriate for the evaluation of sleep disorders like SRBD. The best identification of sleep irregularities at level 89.00 percent for the raw database was obtained. Thus, statistical predictive models allow identification of breathing events with high levels of sensitivity and specificity, providing an inexpensive and accurate diagnosis.     

A simple method for the extraction of soil solution by high speed centrifugation

Summary A simple technique for extracting samples of soil solution by high speed centrifugation is described. The method uses inexpensive commerically available centrifuge tubes requiring only slight modification. Aspects of the method are discussed together with an example of its application.     

A Technique for Continuous Observation of the Infection Process in Cucumber Anthracnose

A new light microscope preparation technique for high magnification observation of living plant tissue and fungal penetration is described. Agar immersion is used in differential interference contrast microscopy (DIC) instead of coverslips (lens 40) or instead of coverslips and oil (lens 100).
This technique is suitable for
(a) longtime observation of living tissue, because the tissue to be observed remains on the plant, and for
(b) thick and uneven samples, as no coverslips are required.
With this technique it was possible to observe the dynamics of penetration of Colletotrichum lagenarium into epideral cells of cucumber cotyledons for 72 hours.
A time lapse film using this technique is in preparation.     

The Helsinki Rat Microsurgical Sidewall Aneurysm Model

Experimental saccular aneurysm models are necessary for testing novel surgical and endovascular treatment options and devices before they are introduced into clinical practice. Furthermore, experimental models are needed to elucidate the complex aneurysm biology leading to rupture of saccular aneurysms.Several different kinds of experimental models for saccular aneurysms have been established in different species. Many of them, however, require special skills, expensive equipment, or special environments, which limits their widespread use. A simple, robust, and inexpensive experimental model is needed as a standardized tool that can be used in a standardized manner in various institutions.The microsurgical rat abdominal aortic sidewall aneurysm model combines the possibility to study both novel endovascular treatment strategies and the molecular basis of aneurysm biology in a standardized and inexpensive manner. Standardized grafts by means of shape, size, and geometry are harvested from a donor rat''s descending thoracic aorta and then transplanted to a syngenic recipient rat. The aneurysms are sutured end-to-side with continuous or interrupted 9-0 nylon sutures to the infrarenal abdominal aorta.We present step-by-step procedural instructions, information on necessary equipment, and discuss important anatomical and surgical details for successful microsurgical creation of an abdominal aortic sidewall aneurysm in the rat.