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PubMed2000

A comparison of optical geometries for combined flash photolysis and total internal reflection fluorescence microscopy.

Total internal reflection fluorescence (TIRF) microscopy, used in conjunction with flash photolysis, provides a useful way of investigating the kinetics of macromolecular interactions. We compare different TIRF optical geometries to establish an optimal combination. Excitation light was introduced via four different arrangements: (1) a prism positioned on the microscope optical axis, (2) an offset prism with propagation through a silica slide trans to the objective lens, (3) an offset prism with propagation through a silica coverslip cis to a water-immersion objective lens and (4) a prismless arrangement using a high NA oil-immersion objective lens. Photolysis was achieved using a Xe flash lamp and a customised silica condenser lens. Single myosin molecules labelled with a Cy3 fluorophore were used as a test sample. Although the offset trans prism gave the best signal-to-background ratio, a customised thin rhombic prism incorporated, on axis, into the flash condenser assembly was almost as good and was more practical for scanning multiple fields. An oil-immersion lens gave the brightest image for sample depths < 30 micrometer but above this limit, a water-immersion lens was better. The prismless arrangement may offer advantages in other situations but it is important to check the actual numerical aperture of the objective lens.

Animals↗
PubMed2003

Flash electroretinography in standing horses using the DTL microfiber electrode.

PURPOSE: The goal of our study was the evaluation of a practical method for the recording of flash electroretinograms (ERGs) in sedated, standing horses with the DTL microfiber electrode. METHODS: The horses were sedated intravenously with detomidine hydrochloride (0.015 mg/kg). The pupil was dilated and the auriculopalpebral nerve was blocked. The ERGs were recorded with the active electrode on the cornea (DTL), the reference electrode near the lateral canthus, and the ground electrode over the occipital bone. The light intensities of the white strobe light were 0.03 cd x s/m2 (scotopic) and 3 cd x s/m2 (scotopic and photopic). Photopic and scotopic single flash and flicker responses to Ganzfeld stimulation were recorded. During the 20-min dark adaptation period the retina was stimulated every 5 min with the 0.03 cd x s/m2 single flash. RESULTS: The median b-wave amplitudes and implicit times were 38 microV and 33 ms (photopic cone-dominated response), 43 microV and 63 ms (5-min dark adaptation), 72 microV and 89 ms (10 min), 147 microV and 103 ms (15 min), 188 microV and 109 ms (20 min, 0.03 cd x s/m2, rod response), and 186 microV and 77 ms (20 min, 3 cd x s/m2, maximal combined rod-cone response). A steady increase in amplitude and implicit time was noted during dark adaptation. No oscillatory potentials could be isolated. CONCLUSIONS: The use of detomidine hydrochloride sedation and the DTL microfiber electrode allowed the recording of good quality ERGs. This protocol should permit the detection of functional problems in the retina without the risk involved with general anesthesia.

Animals↗
PubMed1999

[Optimization of numerical measurement parameters for ECG-triggered MRI snapshot-FLASH myocardial perfusion studies].

BACKGROUND AND AIMS: In MR examinations of myocardial perfusion by the use of Snapshot-FLASH sequences it is of major importance that the achievable signal difference between pre- and normal postcontrast myocardium be maximized. METHODS: In ECG-triggered Snapshot-FLASH sequences the signal intensity of the myocardium depends on the flip angle alpha, the inversion time TI and the trigger delay TD (both depending on the cardiac frequency f) for unchanged slice thickness (SL), matrix size (MA), repetition time (TR) and echo time (TE). Therefore a simulation of the signal behavior of pre- and postcontrast myocardium based on Bloch's equations was performed by varying the flip angle alpha, TI and TD for different cardiac frequencies in order to determine an optimized combination of the measurement parameters. RESULTS: In normal heart rates (50-70 beats/min) maximal signal differences between pre- and normal postcontrast myocardium can be reached for inversion times TI = 170-200 ms and a flip angle alpha = 11 degrees. For higher heart rates again alpha = 11 degrees and TI = 200-220 ms with shortened TD (TD = 0 for f > 90 beats/min) were found to be optimal. The calculated values were semiquantitatively confirmed in phantom and volunteer measurements. CONCLUSIONS: The described method allows cardiac frequency dependent optimization of the Snapshot-FLASH measurement parameters alpha, TI and TD in order to reach a maximum in signal contrast between normal and malperfused myocardium.

Computer Graphics↗
PubMed1996

[Flow contrast behavior in fast MR angiography with an ECG-triggered, slice-selective inversion recovery turbo-FLASH technique].

PURPOSE: To optimise and evaluate fast "inflow"-based contrast-prepared gradient-echo magnetic resonance angiography (MRA) of blood vessels with pulsatile flow. MATERIAL AND METHOD: We used a standard 2-D Turbo-FLASH sequence with an acquisition time of 0.9 s and a slice-selective hyperbolic secant inversion (SSIR) pulse for contrast preparation. Trigger delay and inversion time were systematically varied. RESULTS: Abdominal blood vessels of volunteers showed high signal intensity compared to stationary tissue without pulsatile artifacts. However, depending on cardiac cycle and inversion time, venous vessels in particular exhibited a periodic signal drop by maximum 70% of the peak value. Using optimised parameters SSIR turbo-FLASH images of patients could be obtained with high flow contrast between normal and pathological structures. CONCLUSION: With ECG triggering SSIR Turbo-FLASH MRA enables fast imaging of pulsatile vessels with optimised flow contrast.

Abdomen↗
PubMed1991

[Contrasts in the FLASH sequences in the study of musculoskeletal tumors: phantom study and theoretical calculations].

The contrast between samples simulating musculoskeletal neoplasms and muscle or bone marrow achieved with FLASH sequences, was analysed in phantom studies and was compared with theoretically calculated contrast with and without correction for flip angle distribution over the slice profile. The contrast was correlated closer with the flip angle than with TR or TE. For delineation of the tumour from muscle, only a FLASH sequence with a large flip angle following intravenous administration of Gd-DTPA can be recommended, if a tumour shows a clear Gd-DTPA uptake. With all FLASH sequences analysed, no sufficient contrast between "tumour" without Gd-DTPA uptake and muscle was obtained. Maximal contrast between "tumour" and bone marrow was achieved with small flip angles; and an additional peak was noted with large flip angles and short TR. Experimentally measured T2*-dependent contrasts were nearly identical with theoretically calculated contrasts without correction of flip angle distribution. For calculation of T1-dependent contrasts, correction of the flip angle distribution over the slice profile was of high value.

Bone Neoplasms↗
PubMed1988

[Parameter-weighted MR images and their optimization: theory and example of images for the flash sequence compared to the spin-echo sequence].

In this paper the signal intensity and parameter-weighting zones of spin echo and gradient echo sequences (FLASH) have been analysed. Using information available in the literature, with additional analytical and numerical techniques, it is possible to determine parameters which maximise tissue contrast for T1, T2 and proton-weighted images. The technique is equally applicable to spin echo and FLASH sequences. The advantages of optimised gradient echo sequences compared with spin echo sequences with respect to its signal to noise ratio for short repetition times are analysed quantitatively. T2-weighted images can be generated as rapid as T1-weighted images, using the FLASH sequences when using a small angle of spin and relatively long echo. Illustrations are shown to demonstrate the numerical results (field strength 1.5 tesla). This is followed by a discussion of possible opposing effects.

Humans↗
PubMed1980

[Electromechanical flash stimulator for erg-pev (author's transl)].

In order to avoid electrical interference and the noice caused by a gas discharge tube (flash), the authors have developed a stimulator using mechanical flash as light modulator for ERG and PEV. Flash duration is 2 ms regardless of frequency. This apparatus is now on the market.

Electric Stimulation↗
PubMed1998

Effect of the timing of treatment of port-wine stains with the flash-lamp-pumped pulsed-dye laser.

BACKGROUND: Port-wine stains can be treated with a flash-lamp-pumped pulsed-dye laser, but it is uncertain whether this treatment is more effective if administered early in life, when the skin is thinner and the lesion is smaller. METHODS: We prospectively studied 100 patients with a previously untreated port-wine stain of the head or neck. They were treated with the flash-lamp-pumped pulsed-dye laser and divided into four age groups (0 to 5, 6 to 11, 12 to 17, and 18 to 31 years). The outcome measure was lightening of the port-wine stain (reduction in the difference in color between the skin with the stain and contralateral healthy skin) as measured with a colorimeter after an average of five treatments (range, three to seven) of the entire lesion. RESULTS: Of the 100 patients, 11 could not be included in the analysis because they had received fewer than three or more than seven treatments, had an erroneous base-line color measurement, or were lost to follow-up. The sizes, locations, and colors of the port-wine stains were similar among the groups. When all 89 patients were analyzed together, the average reduction in the difference in color between the skin with the port-wine stain and contralateral healthy skin was 40 percent. The differences between age groups in the average reduction in color differences were not significant (P= 0.26). By the end of the study, only 7 of 89 patients had completed laser therapy, and in no case was clearance complete. Treatment was discontinued in all seven because the last three treatments did not lead to further lightening. CONCLUSIONS: We found no evidence that treatment of port-wine stains with the flash-lamp-pumped pulsed-dye laser in early childhood is more effective than treatment at a later age.

Adolescent↗
PubMed2006

FLASH is an essential component of Cajal bodies.

Cajal bodies are small nuclear organelles with a number of nuclear functions. Here we show that FLICE-associated huge protein (FLASH), originally described as a component of the apoptosis signaling pathway, is mainly localized in Cajal bodies and is essential for their structure. Reduction in FLASH expression by short hairpin RNA results in disruption of the normal architecture of the Cajal body and relocalization of its components. Because the function of FLASH in the apoptosis receptor signaling pathway has been strongly questioned, we have now identified a clear function for this protein.

Animals↗
PubMed1974

The effect of bicarbonate on photosynthetic oxygen evolution in flashing light in chloroplast fragments.

The ability of bicarbonate ion (HCO(3) (-)) to stimulate photosynthetic oxygen evolution in maize chloroplast fragments exposed to continuous light depends on light intensity. Stimulation by HCO(3) (-) is less at low intensities. In HCO(3) (-)-depleted chloroplasts exposed to brief saturating light flashes, period 4 oscillations (in O(2) yield per flash) are damped within three cycles. Readdition of HCO(3) (-) to these preparations restores the oscillatory pattern to higher flash numbers, indicating that HCO(3) (-) reduces the probability of "misses" in the photosystem II reaction center. The rate of the dark relaxation reaction S(n) (') --> S(n+1) (where S refers to the oxidation state of the oxygen-evolving mechanism and n = 0, 1, or 2), after a photoact in the photosystem II reaction center, is retarded in HCO(3) (-)-depleted chloroplasts compared to the rate for this reaction in depleted chloroplasts to which HCO(3) (-) has been resupplied. However, the final oxygen-evolving reaction after the accumulation of four positive charges appears to be independent of HCO(3) (-). Bicarbonate has no effect on the dark deactivation of the higher oxidation states (S(2) and S(3)) of the positive charge-accumulating system. We propose two alternate ways in which the kinetic model of oxygen evolution developed by Kok et al. [(1970) Photochem. Photobiol. 11, 457-475] can be extended to include the action of HCO(3) (-).

Journal Article↗
PubMed1997

Plastoquinol at the quinol oxidation site of reduced cytochrome bf mediates signal transduction between light and protein phosphorylation: thylakoid protein kinase deactivation by a single-turnover flash.

Redox-controlled phosphorylation of thylakoid membrane proteins represents a unique system for the regulation of light energy utilization in photosynthesis. The molecular mechanisms for this process remain unknown, but current views suggest that the plastoquinone pool directly controls the activation of the kinase. On the basis of enzyme activation by a pH shift in the darkness combined with flash photolysis, EPR, and optical spectroscopy we propose that activation occurs when plastoquinol occupies the quinol-oxidation (Qo) site of the cytochrome bf complex, having its high-potential path components in a reduced state. A linear correlation between kinase activation and accessibility of the Qo site to plastoquinol was established by quantification of the shift in the g(y) EPR signal of the Rieske Fe-S center resulting from displacement of the Qo-site plastoquinol by a quinone analog. Activity persists as long as one plastoquinol per cytochrome bf is still available. Withdrawal of one electron from this plastoquinol after a single-turnover flash exciting photosystem I leads to deactivation of the kinase parallel with a decrease in the g(z) EPR signal of the reduced Rieske Fe-S center. Cytochrome f, plastocyanin, and P(700) are rereduced after the flash, indicating that the plastoquinol at the Qo site is limiting in maintaining the kinase activity. These results give direct evidence for a functional cytochrome bf-kinase interaction, analogous to a signal transduction system where the cytochrome bf is the receptor and the ligand is the plastoquinol at the Qo site.

Journal Article↗
PubMed1988

The quantal source of area supralinearity of flash responses in Limulus photoreceptors.

The time integrals of the responses of dark-adapted Limulus ventral photoreceptors to flashes exhibit a supralinear dependence on intensity at intermediate intensities. By decomposing the responses into their elementary single-photon components ("bumps"), we are able to calculate the overall quantum efficiency and to display the time courses of the bump amplitude and rate of appearance. Since the time course of the flash response is not slow compared with that of the bump, it was necessary, in order to carry out the decomposition, to develop a new technique for noise analysis of dynamic signals. This new technique should have wide applications. Our main finding is that the supralinearity of the flash responses corresponds to an increase in bump amplitude, with little change in bump duration or quantum efficiency. The time courses of the bump rate and of the change in bump amplitude are peaked and have widths similar to that of the response itself. The peaks of the time courses of the bump rate and amplitude displayed against the starting times of the bumps do not coincide and occur approximately 80 and approximately 40 ms, respectively, before the peak of the response. The time from the start of a bump to its centroid is approximately 70 ms, which means that the time at which the bump centroid reaches its maximum follows the response peak by 30 ms. These results impose constraints on possible mechanisms for the amplitude enhancement.

Animals↗
PubMed1991

Flash photolysis of magnesium-DM-nitrophen in heart cells. A novel approach to probe magnesium- and ATP-dependent regulation of calcium channels.

Changes in cytoplasmic magnesium concentration in the millimolar range are known to evoke directionally opposite changes in the amplitude of cardiac L-type calcium current, but the effects of magnesium changes in the micromolar range are not known. This concentration range is particularly interesting for two reasons: open-channel block of some ion channels (eg, the inward rectifier) occurs with a micromolar Kd; and magnesium at such low levels may limit the availability of magnesium adenosine triphosphate (MgATP) and magnesium guanidine triphosphate (MgGTP) for a variety of enzymatic processes, including calcium channel phosphorylation. To investigate the effects of critical magnesium depletion and its sudden reversal, we used flash photolysis to liberate magnesium from intracellular DM-nitrophen in guinea pig heart cells. Under our conditions, intracellular [Mg2+] changed from 0.25 mumol/L before the flash to less than or equal to 200 mumols/L afterwards. Calcium channel currents carried by either calcium or barium consistently increased after flash, a directional change opposite to the reported effects of elevated [Mg2+] in the millimolar range. Our data do not yet enable us to understand the basis of the magnesium effect, but we are particularly interested in the possibility that the sudden appearance of magnesium enables calcium channel phosphorylation.

Acetates↗
PubMed1996

First-pass evaluation of myocardial output during dipyridamole stress using turbo-FLASH magnetic resonance imaging.

RATIONALE AND OBJECTIVES: This study evaluated the value of dynamically enhanced fast low-angle shot (FLASH) magnetic resonance (MR) imaging in measuring cardiac output with and without dipyridamole pharmacological stress. METHODS: Ten subjects underwent rest and stress MR imaging. Rest images were acquired using electrocardiogram gated MR (turbo-FLASH: repetition time = 6 mseconds; echo time = 12 mseconds; flip angle = 12 degrees, inversion time = 100) 10 to 45 seconds after intravenous bolus of 0.04 mmol/kg gadolinium (Gd)-DTPA using a Siemens 1.0-tesla Magnetom SP. Stress was induced within the MR imaging scanner with 0.56 mg/kg dipyridamole over 4 minutes with stress MR images obtained after a second bolus of Gd-DTPA in exactly the same position and time intervals. Cardiac output was calculated with a least squares error analysis before and after dipyridamole stress for the left and right ventricles in all 10 patients, and comparison was made with cardiac output by Fick dilution technique during cardiac catheterization in seven patients. RESULTS: This MR analysis methodology shows reasonable correlation (r = 0.953) between left ventricular and right ventricular cardiac output with no effect on cardiac output during immediate dipyridamole stress. Fick dilution studies demonstrated a correlation of 0.96. CONCLUSIONS: Turbo-FLASH MR can demonstrate time-activity curves and cardiac output calculations consistent with theoretical predictions.

Aged↗
PubMedDate not supplied

Three-dimensional FLASH imaging: applications with gadolinium-DTPA.

Sixteen patients with a variety of brain and spine tumors were examined with T1-weighted spin echo and three-dimensional (3D) fast low-angle shot (FLASH) (60 degree flip angle) techniques before and after administration of Gd-diethylenetriamine pentaacetic acid (DTPA) to assess the utility of volume imaging in parenchymal and dural disease. Reconstructions were performed on a Kontron MIP magnetic resonance (MR) workstation and compared with the corresponding spin echo images. Good 3D FLASH examinations (n = 9) were seen with a variety of echo times (6-17 ms) and repetition times (40-100 ms). Fair (n = 4) and poor (n = 3) examinations were usually associated with motion artifact, especially the coronal acquisitions. Spin echo images were all considered of good quality. An increase in confidence of diagnosis was seen in nine cases following contrast medium administration, based on either enhancement of an abnormality (n = 7) or failure of enhancement (n = 2). Three-dimensional FLASH imaging combined with Gd-DTPA holds promise to develop into a clinically valuable sequence allowing good patient throughput and ability to reconstruct in any plane and sensitivity to abnormalities of the blood-brain barrier or vascularity. However, some problems remain, including motion and susceptibility artifacts, long reconstruction time, and loss of resolution in reconstruction due to anisotropic volume.

Brain Neoplasms↗
PubMedDate not supplied

T1 maps by K-space reduced snapshot-FLASH MRI.

The T1 maps evaluated from k-space reduced Snapshot fast low angle shot (FLASH) images provide high contrast parameter images for tissue characterization in vivo of any body region. An algorithm for computing T1 values that allows a fast and reliable evaluation of T1 maps and yields reproducible values of tissue parameters in MR imaging is presented. The algorithm combined with the Snapshot FLASH inversion recovery imaging sequence permits a precise determination of T1 values, even for T1 times as low as 50 ms. Comparison with a spectroscopical inversion recovery method on identical phantoms demonstrates the accuracy of this technique. With its total acquisition time of approximately 2 s, IR Snapshot FLASH is fast enough to be used in monitoring fast T1 dynamics.

Magnetic Resonance Imaging↗
PubMedDate not supplied

Endometrial carcinoma: dynamic MRI with turbo-FLASH technique.

PURPOSE: Our goal was to investigate the usefulness of dynamic MRI with the turbo-FLASH technique in estimating myometrial invasion by endometrial carcinoma. METHOD: Forty-six patients with endometrial carcinomas were evaluated with pathologic correlation. Dynamic MRI was performed with the rapid administration of Gd-DTPA using the turbo-FLASH technique. RESULTS: The inner muscle layer showed more rapid contrast enhancement effects than the outer muscle layer even after menopause. Contrast-to-noise ratio between the inner muscle layer and endometrial carcinoma was maximum at approximately 50 s after administration of Gd-DTPA. In postmenopausal women, the accuracy in estimating myometrial invasion with T2-weighted images, contrast-enhanced T1-weighted images, and dynamic MRI was 66.7, 77.8, and 92.6%, respectively. CONCLUSION: Dynamic MRI with the turbo-FLASH technique is considered to be a useful imaging method for the estimation of myometrial invasion by endometrial carcinoma, especially in postmenopausal patients.

Adult↗
PubMed1986

Multi-flash campimetry as an indicator of visual field loss in glaucoma.

Multi-flash campimetry is a computer-implemented technique used for the rapid assessment of temporal resolving power across the visual field. On the basis of previous reports that glaucoma patients exhibit impaired temporal sensitivity earlier than conventional perimetric field loss, the question of whether multi-flash campimetry might also be sensitive to the visual consequences of glaucoma was explored. Of the 27 patients that were tested, only one had normal temporal resolving power in both eyes, suggesting that multi-flash campimetry might be a useful aid in determining whether a patient should be diagnosed and treated as a glaucoma suspect or as a confirmed glaucoma patient.

Adult↗