Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “dynamic range”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 1,135 records · Page 63Linked to original sources
PubMed1990

Intensity discrimination as the driving force for loudness. Application to pure tones in quiet.

Loudness functions and their associated neural-count functions are derived for steady-state tones at 250 and 1000 Hz from measurements of intensity discrimination obtained under gated and continuous conditions. The calculations are based on a multichannel generalization of the McGill-Goldberg counting model [J. Acoust. Soc. Am. 44, 576-581 (1968)]. Using the data for just noticeable differences (jnd) in intensity as input, the generalized version gives an integral relation between the neural-count function N(x) and the intensity-jnd function, where x = I/I0 and I0 is the reference intensity. Loudness functions are generated through the prescription L(x) = AN(x)--B. To determine how the detailed shapes of the intensity-jnd functions affect the form of the loudness function within the model, integration was performed over the intensity-jnd functions with and without a power-function approximation. Over a range of intensity levels from 20-95 dB, the best agreement between the calculated and measured loudness functions is obtained from the unaltered intensity-jnd functions. Consistent with psychophysical evidence and several models of intensity coding, the results predict that the output of the whole auditory nerve is unnecessary to maintain the large dynamic range observed for loudness and intensity discrimination.

Animals↗
PubMed2001

Temporal representation of iterated rippled noise as a function of delay and sound level in the ventral cochlear nucleus.

The discharge patterns of single units in the ventral cochlear nucleus (VCN) of anesthetized guinea pigs were examined in response to iterated rippled noise (IRN) as a function of the IRN delay (which determines the IRN pitch) and the IRN sound level. Delays were varied over five octaves in half-octave steps, and sound levels were varied over a 30- or 50-dB range in steps of 5 dB. Neural responses were analyzed in terms of first-order and all-order inter-spike intervals (ISIs). The IRN quasi-periodicity was preserved in the all-order ISIs for most units independent of unit type or best frequency (BF). A deterioration of the temporal all-order code was found, however, when the neural response was influenced by inhibition. The IRN quasi-periodicity was also preserved in first-order ISIs for a limited range of IRN delays and levels. Sustained Chopper units (CS) in the VCN responded with very regular ISIs when the IRN delay corresponded to the unit's chopping period; i.e., the unit showed an increased proportion of intervals corresponding to the IRN delay (interval enhancement) relative to an equal-level, white-noise stimulation. This interval enhancement has a band-pass characteristic with a peak corresponding to the chopping period. Moreover, for CS units in rate saturation, the chopping period, and thus the interval enhancement to the IRN, did not vary with level. Units classified as onset-chopper also show a band-pass interval enhancement to the IRN stimuli; however, they show more level-dependent changes than CS units. Primary-like (PL) units also show level-dependent changes in their ability to code the IRN pitch in first-order intervals. The range of delays where PL units showed interval enhancement was broader and extended to shorter delays. Based on these findings, it is suggested that CS units may play an important role in pitch processing in that they transform a higher-order interval code into a first-order interval place code. Their limited dynamic range together with the preservation of the temporal stimulus features in saturation may serve as a physiological basis for the perceived level independence of pitch.

Action Potentials↗
PubMed2006

The six spot step test: a new measurement for walking ability in multiple sclerosis.

OBJECTIVE: The primary objective of this study was to develop a quantitative test to assess ambulation in multiple sclerosis (MS) patients that is more accurate and sensitive than the Timed 25-foot walk (T25FW). For this purpose, we developed the Six Spot Step Test (SSST), which besides speed includes co-ordination and balance, to be a lower limb counterpart to the 9-Hole Peg Test (9HPT). BACKGROUND: The T25FW, which is the ambulation test of the MS Functional Composite (MSFC), reflects only the speed component of walking. The lack of sensitivity to other components of gait adds to the floor effect. METHODS AND PATIENTS: In the SSST, the patient is instructed to walk as quickly as possible from one end to the other of a rectangular field measuring 1 x 5 m, while kicking five cylinder blocks out of five circles marked on the floor. Some 151 MS patients with the Expanded Disability Status Scale (EDSS) score 0-6.5 and 64 normal controls performed the SSST and the T25FW. In addition, 41 patients performed the tests twice. RESULTS: The range of the SSST (4.7-35.1 seconds) was wider than that of the T25FW (3.5-22.6 seconds). Using control mean + 2 SD as cut off, 107 patients had abnormal SSST, while 100 patients had abnormal T25FW. The T25FW (mean) increased 2.1 seconds over the EDSS range of 0-4.0, while the SSST increased 4.9 seconds. The intra-class correlation between repeated tests (r) was 0.95 for the SSST and 0.96 for the T25FW. The correlation between the SSST and the T25FW was high (r=0.92). CONCLUSION: The SSST seems to be superior to the T25FW in terms of dynamic range, floor effect and discriminatory power. The SSST is a relevant alternative for the T25FW as the ambulation component of the MSFC.

Adult↗
PubMed1991

Mammographic equipment, technique, and quality control.

The most important improvements in mammographic technique were the introduction of single- or double-emulsion high-contrast film-screen combinations for mammography, the use of a specially designed low-kilovoltage Bucky grid to reduce scattered radiation, and the introduction of smaller focal spots to improve imaging geometry. Magnification techniques, especially the spot-film technique, yields clearer delineation of high-contrast microcalcifications. Dedicated mammographic equipment with specially designed x-ray tubes is necessary for modern high-quality mammography. However, in many modern mammographic units, the automatic exposure controller still fails to provide appropriate and constant optical film density over a wide range of tissue thickness and absorption. Extended-cycle processing of single-emulsion mammographic films can yield better image contrast and reduce exposure by up to 30%. Exposure times of less than 1 second are recommended to avoid the unnecessary higher doses caused by longer exposure times and reciprocity law failure. The wide dynamic range in mammography can be reduced by a beam equalization filter, and thus be better adapted to the decreased latitude of modern high-contrast mammographic screen-film systems. Mammographic film reading (detection of subtle microcalcifications) can be facilitated by modern computer evaluation of previously digitized mammograms. Standardization and assurance of image quality have been major challenges in the technical development of mammography. Different technical and anthropomorphic phantoms have been designed to measure and compare practical image quality. Detailed quality control measures have been developed. The benefit of a single or annual screening mammography, calculated in gained life expectancy, by far outweighs the relative risk for radiation-induced breast cancer.

Female↗
PubMed1998

[Otoacoustic distortion products in infants and adults: a comparative study].

Distortion Products (DP) are otoacoustic emissions evoked by emitting two pure tones. The principle advantage of this method vs. transient evoked otoacoustic emissions is the specificness of frequency. The purpose of the present work was to determine whether this technique could be applied in objective study of cochlear function in infants. This was done by setting standard data for infants and comparing them with the data obtained in a control population of adults. The input/output functions of the DP and the DP audiograms for 8 f2 frequencies ranging from 696 to 6006 Hz were studied in a group of 15 healthy, full-term infants. The data obtained were statistically compared to the data taken from a control group of 8 normal hearing adults. The infants showed significantly more intense DPs in the intermediate frequencies, with a greater detection threshold and dynamic range. On the other hand, at the high frequencies the amplitude of infant DPs was the same or lower than (f2 = 5005) those found in the adults. These differences can partially be attributed to the fact that the outer auditory channel is smaller in infants thus the probe and the middle ear are better matched. There is significantly greater background noise in infant DPs, particularly at low frequencies. This may be due to the fact that the skin of the outer auditory duct--which acts as a low-pass filter allowing suction and movement of the cervical muscles to contaminate the tracing--is thinner in infants. In conclusion, the study of DPs has shown its potential in the study of cochlear function in infants because it is non invasive and objective, it can be performed quickly and shows frequency specificness. Unfortunately, it cannot be considered a valid method for determining cochlear function at the low frequencies.

Acoustic Stimulation↗
PubMed2005

Bioluminescence regenerative cycle (BRC) system: theoretical considerations for nucleic acid quantification assays.

A novel application of bioluminescence for nucleic acid quantification, the bioluminescence regenerative cycle (BRC), is described in theoretical terms and supported by preliminary experimental data. In the BRC system, pyrophosphate (PPi) molecules are released during biopolymerization and are counted and correlated to DNA copy number. The enzymes ATP-sulfurylase and firefly luciferase are employed to generate photons quantitatively from PPi. Enzymatic unity-gain positive feedback is implemented to amplify photon generation and to compensate for decay in light intensity by self-regulation. The cumulative total of photons can be orders of magnitude higher than in typical chemiluminescent processes. A system level theoretical model is developed, taking into account the kinetics of the regenerative cycle, contamination, and detector noise. Data and simulations show that the photon generation process achieves steady state for the time range of experimental measurements. Based on chain reaction theory, computations show that BRC is very sensitive to variations in the efficiencies of the chemical reactions involved and less sensitive to variations in the quantum yield of the process. We show that BRC can detect attomolar quantities of DNA (10(-18) mol), and that the useful dynamic range is five orders of magnitude. Sensitivity is not constrained by detector performance but rather by background bioluminescence caused by contamination by either PPi or ATP (adenosine triphosphate).

Adenosine Triphosphate↗
PubMed2005

Direct determination of peracetic acid, hydrogen peroxide, and acetic acid in disinfectant solutions by far-ultraviolet absorption spectroscopy.

In this paper we propose a rapid and highly selective far-ultraviolet (FUV) spectroscopic method for the simultaneous determination of peracetic acid (PAA), hydrogen peroxide, and acetic acid (AA). For this purpose we developed a novel FUV spectrometer that enables us to measure the spectra down to 180 nm. Direct determination of PAA, H(2)O(2), and AA, the three main species in disinfectant solutions, was carried out by using their absorption bands in the 180-220-nm region. The proposed method does not require any reagents or catalysts, a calibration standard, and a complicated procedure for the analysis. The only preparation procedure requested is a dilution of H(2)O(2) with pure water to a concentration range lower than 0.2 wt % in the sample solutions. Usually, the required concentration range can be obtained by the 10 times volume dilution of the actual disinfectant solutions. As the measured sample does not leave any impurity for the disinfection, it can be reused completely by using a circulation system. The detection limit for PAA of the new FUV spectrometer was evaluated to be 0.002 wt %, and the dynamic ranges of the measured concentrations were from 0 to 0.05 wt %, from 0 to 0.2 wt %, and from 0 to 0.2 wt % for PAA, H(2)O(2), and AA, respectively. The response time for the simultaneous determination of the three species is 30 s, and the analysis is applicable even to the flowing samples. This method may become a novel approach for the continuous monitoring of PAA in disinfectant solutions on the process of sterilization.

Acetic Acid↗
PubMed2005

Chlorinated phenol analysis using off-line solid-phase extraction and capillary electrophoresis coupled with amperometric detection and a boron-doped diamond microelectrode.

The analysis of chlorinated phenols (2-chlorophenol, 3-chlorophenol, 4-chlorophenol, 2,4-dichlorophenol, 2,4,6-trichlorophenol, pentachlorophenol) in river water was accomplished using off-line solid-phase extraction (SPE) and capillary electrophoresis coupled with electrochemical detection. A key to the sensitive, reproducible, and stable detection of these pollutants was the use of a boron-doped diamond microelectrode in the amperometric detection mode. An off-line SPE procedure was utilized to extract and preconcentrate the pollutants prior to separation and detection, with ENVI-Chrom P, a highly cross-linked styrene-divinylbenzene copolymer, being employed as the sorbent. Pollutant recoveries in the 95-100% range with relative standard deviations of 1-4% were achieved. The diamond microelectrode provided a low and stable background current with low peak-to-peak noise. The oxidative detection of the pollutants was accomplished at +1.05 V vs Ag/AgCl without the need for electrode pretreatment. The method was evaluated in terms of the linear dynamic range, sensitivity, limit of quantitation, response precision, and response stability. A reproducible electrode response was observed during multiple injections of the chlorinated phenol solutions with a relative standard deviation of < or =5.4%. Good electrode response stability was observed over many days of continuous use with no significant electrode deactivation or fouling. The separation efficiencies for all six pollutants were greater than 170,000 plates/m. The minimum concentration detectable for all six ranged from 0.02 to 0.2 ppb (S/N > or = 3) using a 250:1 preconcentration factor.

Boron↗
PubMed1992

Auditory processing of complex sounds: an overview.

The past 30 years has seen a remarkable development in our understanding of how the auditory system--particularly the peripheral system--processes complex sounds. Perhaps the most significant has been our understanding of the mechanisms underlying auditory frequency selectivity and their importance for normal and impaired auditory processing. Physiologically vulnerable cochlear filtering can account for many aspects of our normal and impaired psychophysical frequency selectivity with important consequences for the perception of complex sounds. For normal hearing, remarkable mechanisms in the organ of Corti, involving enhancement of mechanical tuning (in mammals probably by feedback of electro-mechanically generated energy from the hair cells), produce exquisite tuning, reflected in the tuning properties of cochlear nerve fibres. Recent comparisons of physiological (cochlear nerve) and psychophysical frequency selectivity in the same species indicate that the ear's overall frequency selectivity can be accounted for by this cochlear filtering, at least in bandwidth terms. Because this cochlear filtering is physiologically vulnerable, it deteriorates in deleterious conditions of the cochlea--hypoxia, disease, drugs, noise overexposure, mechanical disturbance--and is reflected in impaired psychophysical frequency selectivity. This is a fundamental feature of sensorineural hearing loss of cochlear origin, and is of diagnostic value. This cochlear filtering, particularly as reflected in the temporal patterns of cochlear fibres to complex sounds, is remarkably robust over a wide range of stimulus levels. Furthermore, cochlear filtering properties are a prime determinant of the 'place' and 'time' coding of frequency at the cochlear nerve level, both of which appear to be involved in pitch perception. The problem of how the place and time coding of complex sounds is effected over the ear's remarkably wide dynamic range is briefly addressed. In the auditory brainstem, particularly the dorsal cochlear nucleus, are inhibitory mechanisms responsible for enhancing the spectral and temporal contrasts in complex sounds. These mechanisms are now being dissected neuropharmacologically. At the cortical level, mechanisms are evident that are capable of abstracting biologically relevant features of complex sounds. Fundamental studies of how the auditory system encodes and processes complex sounds are vital to promising recent applications in the diagnosis and rehabilitation of the hearing impaired.

Animals↗
PubMed2006

Bis(2-hydroxyacetophenone)ethylenediimine as a neutral carrier in a coated-wire membrane electrode for lead(II).

A coated-wire ion-selective electrode (CWISE), based on a Schiff base as a neutral carrier, was successfully developed for the detection of Pb(II) in aqueous solution. CWISE exhibited a linear response with a Nernstian slope of 29.4 +/- 0.5 mV/decade within the concentration range of 1.0 x 10(-5) - 1.0 x 10(-1) M lead ion. CWISE has shown detection limits of 5.0 x 10(-6) M. The electrode exhibited good selectivity over a number of alkali, alkaline earth, transition and heavy metal ions. This sensor yielded a steady potential within 10 to 20 s at a linear dynamic range. The electrode was suitable for use in aqueous solutions in a pH range of 2.0 to 5.0. Applications of this electrode for the determination of lead in real samples and as indicator electrode for potentiometric titration of Pb2+ ion using K2CrO4 are reported.

Journal Article↗
PubMed2001

Capillary sodium dodecyl sulfate-DALT electrophoresis of proteins in a single human cancer cell.

Capillary Sodium dodlecyl sulfate (SDS)-DALT an (abbreviation for Dalton) electrophoresis was applied to analysis of proteins in single HT29 human colon adenocarcinoma cells. A vacuum pulse was employed to introduce a single cell into the coated capillary. Once the cell was lysed, proteins were denatured with SDS, fluorescantly labeled with 3-(2-furoyl)-quinoline-2-carboxaldehyde (FQ), and then separated by using 8% pullulan as the sieving matrix. This method offers a few advantages for single-cell protein analysis. First, it provides reproducible separation of single-cell proteins according to their size. Based on comparison with the migration time of standard proteins, most components from a single HT29 cancer cell have molecular masses within the range of 10-100 kDa. Second, as a one-dimensional separation method, it gives fairly good resolution for proteins. Typically, around 30 protein components of a single HT29 cell were resolved, indicating that this method has similar peak capacity to SDS-polyacrylamide gel electrophoresis (PAGE). Third, this method shows high detection sensitivity and wide dynamic range, which is important because of the wide range of protein expression in living systems. Detection limits for standard proteins ranged from 10(-10) to 10(-11) M. Finally, this method provides much higher speed than classical gel electrophoresis methods, and it provides automated anlysis of cellular proteins at the single-cell level; the separation is complete in 30 min and the entire analysis takes approximately 45 min.

Adenocarcinoma↗
PubMed2024

Secretome Analysis Using Affinity Proteomics and Immunoassays: A Focus on Tumor Biology.

The study of the cellular secretome using proteomic techniques continues to capture the attention of the research community across a broad range of topics in biomedical research. Due to their untargeted nature, independence from the model system used, historically superior depth of analysis, as well as comparative affordability, mass spectrometry-based approaches traditionally dominate such analyses. More recently, however, affinity-based proteomic assays have massively gained in analytical depth, which together with their high sensitivity, dynamic range coverage as well as high throughput capabilities render them exquisitely suited to secretome analysis. In this review, we revisit the analytical challenges implied by secretomics and provide an overview of affinity-based proteomic platforms currently available for such analyses, using the study of the tumor secretome as an example for basic and translational research.

Humans↗
PubMed2006

Parameter estimation of feedback gain in a stochastic model of renal hemodynamics: differences between spontaneously hypertensive and Sprague-Dawley rats.

Proximal tubular pressure shows periodic self-sustained oscillations in normotensive rats but highly irregular fluctuations in spontaneously hypertensive rats (SHR). Although we have suggested that the irregular fluctuations in SHR represent low-dimensional deterministic chaos in tubuloglomerular feedback (TGF), they could also arise from other mechanisms, such as intrinsic instabilities in preglomerular vessels or inputs from neighboring, coupled nephrons. To test this possibility, we applied a parameter estimation procedure to a model of TGF, where a stochastic process was added to represent mechanisms not included explicitly in the model. In its deterministic version, the model can have chaotic dynamics arising from TGF. The model introduces random fluctuations into a parameter that determines the gain of TGF. The model shows a rich variety of dynamics ranging from low-dimensional deterministic oscillations and chaos to high-dimensional random fluctuations. To fit the data from normotensive rats, the model must introduce only a small variation in the feedback gain, and its estimates of that gain agree well with experimental values. These results support the use of the deterministic model of nephron dynamics in normotensive rats. In contrast, the irregular tubular pressure fluctuations in SHR were best described by a model dominated by random parameter fluctuations. The results point to the failure of simple mathematical models of nephron dynamics adequately to describe processes that are important for the irregular tubular pressure fluctuations and the need to consider other factors, such as differences in vascular function or nephron-nephron interactions, in further work on this problem.

Animals↗
PubMed1991

Intracellular study of electrophysiological features of primate spinothalamic tract neurons and their responses to afferent inputs.

1. Intracellular recordings were made from 43 spinothalamic (STT) neurons in the lumbosacral region of the spinal cord in anesthetized macaque monkeys. The antidromic responses of these neurons to electrical stimulation of the ventral posterior lateral (VPL) nucleus of the thalamus were examined, and orthodromic responses to electrical stimulation of the sural nerve or to mechanical stimulation of hindlimb skin were recorded to study the electrophysiological features of these neurons and their responses to afferent inputs. 2. The resting membrane potential of the neurons ranged from -26 to -70 mV and the antidromic latency from 2.3 to 9.1 ms. Three of the neurons were located in lamina 1 and were recorded so briefly that only antidromic and spontaneous activity could be studied. The rest of the neurons were located in laminae III-V and were of the wide-dynamic-range (WDR) type. 3. The antidromic action potentials recorded in the somas of STT neurons typically showed a fast rising phase and a short initial segment-somadendritic (IS-SD) delay. After repetitive antidromic stimulation, a progressive elongation of the IS-SD delay, widening of the spike, and failure of the SD spike were observed. 4. The afterpotential of the antidromic action potential consisted of a fast afterhyperpolarization (AHPf) and sometimes a delayed depolarization (DD) and a slow afterhyperpolarization (AHPs). The amplitude and the duration of the AHPs were progressively increased when longer trains of stimuli were used. When the membrane potential was hyperpolarized, the amplitude of the AHPs decreased, suggesting an involvement of K+ and/or Cl- ions. However, the AHPs completely disappeared when the strength of stimulation was adjusted to a level just below the threshold for the axon, suggesting that it was unlikely that recurrent inhibition contributed to the AHPs. 5. The background activity of 32 STT neurons was analyzed. The membrane potential at which spikes were triggered in these neurons was around -42 mV. The width and the rise time of the spontaneous spikes were shorter than those of antidromic action potentials, although the maximum rate of rise was similar. The heights of the spontaneous spikes were slightly shorter than those of antidromic action potentials. 6. Three types of background activity have been observed. One type had a very low average spontaneous rate with a bursting firing pattern, consisting of a few spikes superimposed on a depolarization. This type of activity was seen mostly in lamina I neurons. The second type of activity had a moderate to high spontaneous rate with a fairly constant interval between spikes.(ABSTRACT TRUNCATED AT 400 WORDS)

Action Potentials↗
PubMedDate not supplied

Functional biomechanical deficits in running athletes with plantar fasciitis.

Plantar fasciitis is a relatively common injury that occurs in running athletes. The disease entity is a good example of an overloaded process of the plantar fascia at its calcaneal insertion. This study was designed to examine the strength and flexibility findings in the muscles that are put on tensile load during running, and which are responsible for controlling the forces on the foot during stance and pushoff, thus modifying the overload. Three groups of athletes underwent physical examination, including checking ankle range of motion in plantar flexion and dorsiflexion. Cybex peak torque measurements were taken at 60 and 180 deg/sec. The groups were a control group of 45 athletes with no symptoms, a group that included 43 affected feet with symptomatic plantar fasciitis, and a group that included the 43 unaffected contralateral feet. Analysis of data showed dynamic range of motion deficits in 38 of 43 affected feet, static range of motion deficits in 37 of 43 affected feet, deficits in peak torque at 60 deg/sec in 41 of 43 affected feet, and deficits in peak torque at 180 deg/sec in 37 of 43 affected feet. Statistical comparison of range of motion showed that the group with symptomatic plantar fasciitis was significantly restricted compared to both control and unaffected contralateral feet groups. Statistical comparison of peak torque showed that the symptomatic plantar fasciitis group was significantly lower than both other groups at both velocities. This study documents strength and flexibility deficits in the supporting musculature of the posterior calf and foot that are affected by plantar fasciitis.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed2002

[Design and evaluation of a confocal laser-induced fluorescence detector].

A portable laser-induced fluorescence detector, based on confocal configuration detection system has been developed. This is assembled from commercially available components. All the components of the detector are domestic, which makes it low cost. The routine alignment procedure is simplified by using a skillful and visual alignment system and requires minimal experience for operation. The module design makes it possible for high performance liquid chromatographic, capillary electrophoretic and microfluid chip applications. The performance of the detector, including the sensitivity, noise, linear range and detection limit, was evaluated by capillary electrophoresis and flow injection analytical technique using a red-absorbing cyanine derivative (Cy5) and Cy5 labeled tryptophan as test samples. The results show that the background signal is very low and the peak-to-peak noise level is 0.002 mV. The detection limit and the linear dynamic range are 3.7 nmol/L and 10(3), respectively.

Chromatography, High Pressure Liquid↗
PubMedDate not supplied

Application of a new high sensitivity luminometer for industrial microbiology and molecular biology.

A compact new luminometer (FB12) has been developed based on a 370-630 nm photon counter and measuring chamber that can accommodate a range of sample formats. The FB12 permits measurements as low as 1000 molecules of luciferase in reporter gene assays. Its sensitivity for ATP is limited by reagent background. If ATP assay reagents had no chemical background, 2 fg of ATP could be detected using 3 SD of instrument background as the detection limit. The FB12 has a dynamic range of six decades and operates under its own microprocessor programme or protocol-based PC software that is integrated with Microsoft(R) Excel(R). An injector port above the sample measuring position allows connection of external reagent injectors. Applications are performed using protocols provided with the FB12 or user defined protocols. Examples are presented that illustrate use of the instrument for research and industrial applications.

Industry↗
PubMed2001

A real-time ratiometric method for the determination of molecular oxygen inside living cells using sol-gel-based spherical optical nanosensors with applications to rat C6 glioma.

The first sol-gel-based, ratiometric, optical nanosensors, or sol-gel probes encapsulated by biologically localized embedding (PEBBLEs), are made and demonstrated here to enable reliable, real-time measurements of subcellular molecular oxygen. Sensors were made using a modified Stöber method, with poly(ethylene glycol) as a steric stabilizer. The radii of these spherical PEBBLE sensors range from about 50 to 300 nm. These sensors incorporate an oxygen-sensitive fluorescent indicator, Ru(II)-tris(4,7-diphenyl-1,10-phenanthroline) chloride ([Ru(dpp)3]2+), and an oxygen-insensitive fluorescent dye, Oregon Green 488-dextran, as a reference for the purpose of ratiometric intensity measurements. The PEBBLE sensors have excellent reversibility, dynamic range, and stability to leaching and photobleaching. The small size and inert matrix of these sensors allow them to be inserted into living cells with minimal physical and chemical perturbations to their biological functions. Applications of sol-gel PEBBLEs inserted in rat C6 glioma cells for real-time intracellular oxygen analysis are demonstrated. Compared to using free dyes for intracellular measurements, the PEBBLE matrix protects the fluorescent dyes from interference by proteins in cells, enabling reliable in vivo chemical analysis. Conversely, the matrix also significantly reduces the toxicity of the indicator and reference dyes to the cells, so that a wide variety of dyes can be used in optimal fashion.

Animals↗