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At least 1,747 records · Page 97Linked to original sources

Micromachining in plastics using X-ray lithography for the fabrication of microelectrophoresis devices.

Micromachining was performed in polymethylmethacrylate (PMMA) using X-ray lithography for the fabrication of miniaturized devices (microchips) for potential applications in chemical and genetic analyses. The devices were fabricated using two different techniques: transfer mask technology and a Kapton mask. For both processes, the channel topography was transferred (1:1) to the appropriate substrate via the use of an optical mask. In the case of the transfer mask technique, the PMMA substrate was coated with a positive photoresist and a thin Au/Cr plating base. Following UV exposure, the resist was developed and a thick overlayer (approximately 3 microns) of Au electroplated onto the PMMA substrate only where the resist was removed, which acted as an absorber of the X-rays. In the other technique, a Kapton film was used as the X-ray mask. In this case, the Kapton film was UV exposed using the optical mask to define the channel topography and following development of the resist, a thick Au overlayer (8 microns) was electrodeposited onto the Kapton sheet. The PMMA wafer during X-ray exposure was situated directly underneath the Kapton mask. In both cases, the PMMA wafer was exposed to soft X-rays and developed to remove the exposed PMMA. The resulting channels were found to be 20 microns in width (determined by optical mask) with channel depths of approximately 50 microns (determined by x-ray exposure time). In order to demonstrate the utility of this micromachining process, several components were fabricated in PMMA including capillary/chip connectors, injectors for fixed-volume sample introduction, separation channels for electrophoresis and integrated fiber optic fluorescence detectors. These components could be integrated into a single device to assemble a system appropriate for the rapid analysis of various targets.

Electrophoresis, Capillary↗

From scattering to wavefronts--what's in between?

PURPOSE: Some optical errors are too localized and random to be detected by commercial wavefront devices and Zernike polynomial expression. We looked beyond aberrations defined by Zernike expression to discuss implications of fine irregularities associated with highly aberrated corneal surfaces and complex surface roughness that can lead to light scattering. METHODS: Most fine irregularities are related to postoperative surface roughness, complexities of corneal ablation, and the laser in situ keratomileusis (LASIK) flap. These can be characterized mathematically by a random function that includes local surface tilts, the correlation radius of irregularities (Ic), surface roughness, and other terms. The Kirchoff method of scatter analysis characterizes fine surface irregularities by replacing each point on the surface with a tangential plane, allowing it to be governed by Snellen and Fresnel laws. RESULTS: The joint action of the continuum of microbeams defines a complex point spread function that can be expressed by the Strehl ratio. Small, highly irregular steep central islands and flap striae may not be adequately detected by Zernike expression and may have a surface irregularity diameter of 0.1 to 2.0 mm and height of 10 to 20 microm that results in a reduced Strehl ratio below 0.8. Laser ablation inhomogeneities may have dimensions of 1 to 10 microm, resulting in a root mean square tilt value approaching 1.0 and a Strehl ratio below 0.5. CONCLUSION: Corneal surface irregularities after laser vision correction may induce significant optical aberrations and distortions apart from classical wavefront or scattering errors. As these may not be detected by commercial wavefront devices, and yet contribute to the degradation of optical performance, alternate techniques should be evaluated to detect and describe these surface irregularities.

Cornea↗

Single particle high resolution spectral analysis flow cytometry.

BACKGROUND: While conventional multiparameter flow cytometers have proven highly successful, there are several types of analytical measurements that would benefit from a more comprehensive and flexible approach to spectral analysis including, but certainly not limited to spectral deconvolution of overlapping emission spectra, fluorescence resonance energy transfer measurements, metachromic dye analysis, free versus bound dye resolution, and Raman spectroscopy. METHODS: Our system utilizes a diffraction grating to disperse the collected fluorescence and side-scattered light from cells or microspheres passing through the interrogation region over a rectangular charge-coupled-device image sensor. The flow cell and collection optics are taken from a conventional flow cytometer with minimal modifications to assure modularity of the system. RESULTS: Calibration of the prototype spectral analysis flow cytometer included wavelength characterization and calibration of the dispersive optics. Benchmarking of the system demonstrated a single particle/cell intensity sensitivity of 2160 MESF of R-Phycoerythrin. Single particle spectra taken with our instrument were validated against bulk solution fluorimeter and conventional flow cytometer measurements. Coefficients of variation of integrated spectral fluorescence intensity of several sets of standard fluorescent microspheres ranged from 1.4 to 4.8% on the spectral system. Spectral discrimination of free versus PI bound to cells is also demonstrated. CONCLUSIONS: It is demonstrated that the flow spectrometer has sufficient sensitivity and wavelength resolution to detect single cells and microspheres, including multi-fluorophore labeled microspheres. The capability to use both standard mathematical deconvolution techniques for data analysis, coupled with the feasibility of integration with existing flow cytometers, will improve the accuracy and precision of ratiometric measurements, enable the analysis of more discrete emission bands within a given wavelength range, and allow more precise resolution of the relative contribution of individual fluorophores in multiply-tagged samples, thereby enabling a range of new applications involving the spectral analysis of single cells and particles.

Calibration↗

Fiberoptic bronchoscopy during noninvasive positive pressure ventilation delivered by helmet.

OBJECTIVE: To evaluate the feasibility and safety of fiberoptic bronchoscopy (FOB) with bronchoalveolar lavage (BAL) during noninvasive positive pressure ventilation (NPPV) delivered by helmet in patients with acute respiratory failure (ARF) and suspected pneumonia. DESIGN AND SETTING: Prospective, clinical investigation in a general intensive care unit (ICU) of a university hospital. PATIENTS AND PARTICIPANTS: Four adult patients with ARF who underwent NPPV via the helmet and required fiberoptic BAL for suspected pneumonia. INTERVENTIONS: NPPV was delivered through the helmet in the pressure support ventilation mode. The specific seal connector placed in the plastic ring of the helmet allowed the passage of the bronchoscope, maintaining assisted ventilation. Arterial blood gas levels, pH, oxygen saturation, respiratory rate, heart rate, and mean arterial blood pressure were monitored during the study. RESULTS: Helmet NPPV avoided gas exchanges deterioration during FOB and BAL, with good tolerance. During the procedure heart rate increased by 5% and mean arterial blood pressure by 7% over baseline; these levels returned to prebronchoscopic values immediately after the withdrawal of the bronchoscope. Endotracheal intubation was never required during the 24 h after the procedure. BAL yielded diagnostic information in three of four patients. CONCLUSIONS: NPPV through the helmet allows a safe diagnostic FOB with BAL in patients with hypoxemic ARF, avoiding gas exchange deterioration, and endotracheal intubation. ELECTRONIC SUPPLEMENTARY MATERIAL: is available if you access this article at http://dx.doi.org/10.1007/s00134-002-1554-5. On that page (frame on the left side), a link takes you directly to the supplementary materials.

Adult↗

Carbon nanotube transistors for biosensing applications.

Electronic detection of biomolecules, although still in its early stages, is gradually emerging as an effective alternative to optical detection methods. We describe field effect transistor devices with carbon nanotube conducting channels that have been developed and used for biosensing and biodetection. Both transistors with single carbon nanotube conducting channels and devices with nanotube network conducting channels have been fabricated and their electronic characteristics examined. The devices readily respond to changes in the environment, and such effects have been examined using gas molecules and coatings with specific properties. Device operation in (conducting) buffer and in a dry environment--after buffer removal--is also discussed. Applications in the biosensing area are illustrated with three examples: the investigation of the interaction between devices and biomolecules, the electronic monitoring of biomolecular processes, and attempts to integrate cell membranes with active electronic devices.

Animals↗

[Multifocal ERG with confocal scanning laser ophthalmoscope. Comparison with monitor simulation].

BACKGROUND: Multifocal electroretinograms (mfERG) were recorded using a confocal scanning laser ophthalmoscope (cSLO) and compared to the results from conventional monitor stimulation. METHODS: Single and repeated measurements were recorded from 23 normal subjects using the cSLO (Heidelberg Retina Angiograph, Heidelberg Engineering, Heidelberg) as well as a conventional monitor as stimulation devices. Laser power output was modified by various optical filters. The reliability of the method and agreement with the conventional monitor stimulation were determined. RESULTS: CSLO recordings showed a high degree of variability. Reduction of laser power output improved the retinal response topography and characteristically modified response variations with each filter. Differences in amplitude size between cSLO and monitor recordings decreased with increasing amplitude levels. The results of repeated measurements showed considerable variation. CONCLUSION: It is possible to use a cSLO as a stimulator for mfERG recordings. However, a relatively high degree of variability represents a significant limitation of this method. Appropriate reduction of laser power decreases variations and serves to obtain photopic response topographies.

Adult↗

A novel video technique for visualizing flow structures in cardiovascular models.

We describe a video system that produces good quality images of particle trajectories in seeded fluid flows. The operation of a liquid crystal optical shutter and a modified charge-coupled device (CCD) camera were synchronized to generate images of particle trajectories which were stored in a framegrabber before being transferred to S-VHS tape. The camera system is particularly appropriate for visualizing transient or unsteady flows in models of the cardiovascular system as the integration time may be varied to produce particle trajectories of variable length.

Cardiac Output↗

Effect of corneal polarization axis on assessment of retinal nerve fiber layer thickness by scanning laser polarimetry.

PURPOSE: Scanning laser polarimetry uses an anterior segment compensating device that assumes a fixed axis of corneal birefringence, which we call the corneal polarization axis. The purpose of this investigation was to establish the distribution of corneal polarization axes among a population of normal eyes and to evaluate the relationship between corneal polarization axis and posterior segment retardation. METHODS: We constructed a noninvasive slit lamp-mounted device incorporating two crossed linear polarizers and an optical retarder in order to measure the slow axis of corneal birefringence. Normal subjects underwent corneal polarization axis measurement. A subset of eyes underwent scanning laser polarimetry of the peripapillary retinal nerve fiber layer (n = 32) and macula (n = 29), and retardation measurements were evaluated in each group. RESULTS: One hundred eighteen eyes of 63 normal subjects (35 female, 28 male) underwent corneal polarization axis measurement (mean age, 45.5 +/- 17.1 years). Six eyes (5.1%) demonstrated unmeasurable corneal polarization. In the remaining 112 eyes, the mode of the corneal polarization axis distribution was 10 to 20 degrees nasally downward (range, 90 degrees nasally downward to 54 degrees nasally upward). A significant (P <.0001) correlation was observed between fellow eyes (R(2) =.52), with a mean difference of 11.2 +/- 10.5 degrees (range, 0-52 degrees). Corneal polarization axis was significantly associated (R(2) =.52-.84) with retinal nerve fiber layer and macula summary retardation parameters (average thickness, ellipse average, superior and inferior average, superior and total integral; P <.0001 for all groups). CONCLUSIONS: The mean corneal polarization axis among normal corneas is nasally downward; however, considerable intraindividual and interindividual variability exists. The linear relationship between corneal polarization axis and posterior segment retardation parameters is responsible, in part, for the wide distribution of retinal nerve fiber layer thickness data generated by scanning laser polarimetry.

Adult↗

Correction for corneal polarization axis improves the discriminating power of scanning laser polarimetry.

PURPOSE: Corneal polarization axis (CPA) has been reported to affect retardation measurements obtained with scanning laser polarimetry (SLP). The purpose of this investigation was to prospectively determine whether correction for CPA improves the discriminating power of SLP for detection of mild-to-moderate glaucoma. DESIGN: Cross-sectional analysis of normal and glaucomatous eyes. METHODS: We constructed a noninvasive slit-lamp-mounted device incorporating two crossed linear polarizers and an optical retarder to measure the slow axis of corneal polarization. Complete ocular examination, standard automated perimetry, SLP imaging, and CPA measurements were performed on normal and glaucomatous eyes. One eye/subject was enrolled; if both eyes of a patient were eligible for the study, the right eye was selected. For each of the 13 SLP parameters, logistic regression was used to determine if including CPA in the model influenced the ability to discriminate between normal and glaucomatous eyes. RESULTS: Forty-three normal eyes (average visual field mean defect, -0.53 +/- 1.4 dB) and 33 glaucomatous eyes (average visual field mean defect, -5.93 +/- 6.5 dB) were enrolled. CPA was significantly correlated with summary retardation parameters (average thickness and integral values) in normal (r = 0.72-0.83, P <.001 for all values) and glaucomatous eyes (r = 0.43-0.62, P =.013 to <.001). Including CPA in the model improved the ability to discriminate between normal and glaucomatous eyes for five retardation parameters quantifying retinal nerve fiber layer (RNFL) thickness (range of P values: 0.045-0.001). For inferior average thickness, area under the receiver operating characteristic (ROC) curve increased significantly (P =.002) from 0.70 to 0.78 after accounting for CPA; with a sensitivity set at 80% specificity improved from 33% to 72%. Correlations between visual field corrected pattern standard deviation and average thickness, ellipse average, superior average, and inferior average significantly increased (range of P values,.018-.001) after adjustment for CPA (r = -0.35 and -0.45, -0.38 and -0.47, -0.46 and -0.57, and -0.42 and -0.49, respectively). CONCLUSIONS: Correction for CPA significantly increases the correlation between retinal nerve fiber layer structural damage and visual function and significantly improves the discriminating power of SLP for detection of mild-to-moderate glaucoma.

Adult↗

The influence of chamber characteristics on the reliability of sperm concentration and movement measurements obtained by manual and videomicrographic analysis.

To assess the influence of chamber design and depth on the accuracy and precision of sperm measurements, manual counting of a standardized latex bead solution and automated sperm motility measurements were made using a Makler chamber (Sefi Medical Industries, Haifa, Israel), Neubauer hemocytometer (American Optical Company, Buffalo, NY), and a new, disposable device (Micro-Cell; Cyto Fluidics, Inc., Silver Spring, MD). Bead counts obtained with the Micro-Cell chamber or hemocytometer were not statistically different from those determined by electronic particle counting, whereas Makler chamber counts were 62% higher. Makler counts had a significantly higher standard deviation, suggesting that counts made with this device are less reproducible. Analyzing live sperm samples, the percentage of motile sperm determined using Micro-Cell and Makler chambers were similar. However, significant differences in sperm concentration and mean velocity were found. The Micro-Cell disposable chamber provided consistent and accurate data on sperm concentration, percent motility, and mean velocity. These differences in sperm measurements emphasize the importance of sampling chamber characteristics on data reliability.

Cell Survival↗

GRGDSP peptide-bound silicone membranes withstand mechanical flexing in vitro and display enhanced fibroblast adhesion.

Mechanobiological studies of cardiac tissue require devices that allow forces to be exerted on cells in vitro. Silicone elastomer is often used in these devices because it is flexible and transparent, permitting optical imaging of the cells. However, native untreated silicone is hydrophobic and is unsuitable for cell culture. Peptides covalently bound to silicone surfaces are examined here for the enhancement of cellular adhesion during in vitro dynamic flexing. A procedure is described for the chemical modification of medical grade silicone membranes with covalently bound GRGDSP peptides. The conditions for mechanical studies of cardiac cell cultures are then duplicated and it is demonstrated that the peptide layers survive 48 h of mechanical flexing in vitro. Specifically, mechanical flexing in vitro of the 30 pmol/cm2 peptide-modified silicone membranes has no significant effect on the amount of peptides that remains bound to the surface. Cardiac fibroblasts display enhanced adhesion to these peptide-bound silicone membranes for at least 24 h of growth, compared with native silicone or tissue culture polystyrene. The effects of serum versus serum-free media on fibroblast growth are also examined.

Animals↗

Stability of corneal polarization axis measurements for scanning laser polarimetry.

OBJECTIVE: Corneal polarization axis (CPA) has been reported to affect retardation measurements obtained with scanning laser polarimetry. The purpose of this investigation was to evaluate the longitudinal stability of CPA measurements. DESIGN: Prospective, noncomparative case series. PARTICIPANTS: Persons with normal corneas were enrolled; eyes with less than 1 year of follow-up from the initial CPA measurement were excluded. INTERVENTION: We constructed a noninvasive slit-lamp-mounted device incorporating two crossed linear polarizers and an optical retarder to measure the slow axis of corneal birefringence, or CPA. MAIN OUTCOME MEASURES: Corneal polarization axis measurements. RESULTS: Seventy-one eyes of 40 individuals (23 female, 17 male) were enrolled in this investigation (mean age, 42.9 +/- 13.6 years; range, 22-85 years). Initial CPA measurements (mean, 24.0 +/- 18.0 degrees nasally downward; range, 67 degrees downward to 13 degrees nasally upward) were strongly associated (R2 = 0.88; P < 0.0001) with repeat CPA measurements (mean, 20.9 +/- 14.6 degrees nasally downward; range, 59 degrees nasally downward to 14 degrees nasally upward). The mean change in CPA was 4.1 +/- 3.2 degrees (range, 0-13 degrees ). Corneal polarization axis stability was statistically associated with the mean (initial and repeat) CPA (R2 = 0.1; P = 0.009), but not associated with age (R2 = 0.0003; P = 0.9) or gender (R2 = 0.03; P = 0.2). CONCLUSIONS: Corneal polarization axis measurements have good 1-year stability. These data suggest that CPA should not contribute significantly to longitudinal measurements of retinal nerve fiber layer thickness obtained with scanning laser polarimetry.

Adult↗

Devic disease: clinical course, pathophysiology, and management.

Devic disease is characterized by an attack or attacks of optic neuritis and necrotizing myelitis without clinical or magnetic resonance imaging (MRI) evidence of brain involvement or presence of oligoclonal bands in the cerebrospinal fluid (CSF). More than a century after its first systematic study by the French physician, Eugene Devic, its cause as well as its pathogenesis, remains unknown. Devic disease is both a diagnostic and therapeutic challenge to clinicians. The authors present the latest review on the nature, course, possible mechanisms, and therapeutic options of Devic disease.

Journal Article↗

[Orthopedic treatment of scoliosis: new technique using impression by optic procedure].

The authors present a new procedure for acquiring the whole external trunk shape. The construction of braces starts with corrected a positive mold using computer-assisted design software connected to a digital tooling machine. The computer-assisted design and construction of braces enable the determination, measurement, and modification of a three-dimensional image of the trunk, which allows the positive to be corrected. The outer-trunk can be re-balanced, derotated or rectified in the sagittal or frontal plane and the inner modeling of the spine and chest can be rectified. Moreover, in the case of scoliosis or kyphosis, the software produces automatic correction. This rapid (acquisition time < 2 seconds), non invasive, safe and painless procedure can also be used to detect and follow mild spinal deformities.

Adolescent↗

Observer performance comparison of digital radiograph systems for stereotactic breast needle biopsy.

RATIONALE AND OBJECTIVES: Two digital radiograph systems for stereotactic mammography, one using a lens to couple a Lanex Regular screen to a back-illuminated charge-coupled device (CCD) and one using a fiber-optic taper to couple a Min-R Regular-type screen to a front-illuminated CCD, were evaluated with respect to observer performance. METHODS: A contrast-detail phantom was imaged in a variety of equivalent exposure conditions on both systems. Six observers viewed images on a video monitor and recorded which objects were detected. RESULTS: Performance (percent correct detections) with the lens-coupled system using the Lanex Regular screen was significantly higher than with the fiber-optic-coupled system using the Min-R Regular-type screen. CONCLUSION: Differences in absorption efficiencies of phosphors used, as well as differences in design of the two cameras, can explain differences in observer detection performance.

Biopsy, Needle↗

Chemoselective biosensors.

New opportunities for biosensors are now appearing in clinical and genetic diagnostics, genomics, environmental protection, food processing and safety, drug discovery and bioprocess monitoring. Concerns about the cost, stability and selectivity of previous sensor technologies are being addressed by developing new recognition systems and their integration into transducers, micro- and nanofabricated devices, array technologies and novel magnetic, acoustic and optical transduction systems.

Biosensing Techniques↗

Repetitive, noninvasive imaging of cyclooxygenase-2 gene expression in living mice.

The cyclooxygenase-2 (COX-2) gene plays a role in a wide variety of normal physiologic pathways and is a major target of pharmacologic intervention in a large number of pathophysiologic contexts, including pain, fever, inflammation, and cancer. Expression of the COX-2 gene is induced in a wide range of cells, in response to an ever-increasing number of stimuli. The regulation of the COX-2 gene has been the subject of extensive study, using traditional transfection techniques with reporter gene constructs. Regulation of the COX-2 gene in living animals, however, requires sacrifice of the animal and in situ hybridization and/or immunohistochemical studies. We have utilized in vivo optical imaging technology with a cooled charged coupled device camera to image the expression of the firefly luciferase gene in tumor xenografts that are stably transfected with a chimeric gene containing the first kilobase of the murine COX-2 promoter. Induction of luciferase gene expression following systemic lipopolysaccharide/endotoxin administration can be robustly demonstrated; both a dose-response relationship and a time course for luciferase expression from the COX-2 promoter can be noninvasively analyzed in the tumor xenografts. These data suggest expression from the COX-2 promoter will be easily analyzed in transgenic mice, in knock-in mice, and in somatic cell and gene transfer experiments.

Animals↗