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PubMed1992

Role of immersion refractometry for investigating laser-induced effects in cells.

The broad background of scattered light observed in spectra of cell suspensions is reduced by factors of up to 20 by immersion refractometry allowing for improved spectroscopic determination of the absorption properties of cells in the 325-820 nm range. Refractive-index matched spectra of E. coli C1a exhibit a set of resonant features near 422, 561, and 582 nm. Exposure wavelengths are chosen based on this spectrum and cell viability is investigated in E. coli suspensions exposed to 350, 400, 422, 440, and 700 nm radiation delivered in nanosecond pulses with total doses from 500 millijoules to 60 Joules. We observe a loss in cell viability for doses greater than 1 Joule at 422 nm and for all doses at other wavelengths; exposures of less than 1 Joule at 422 nm enhance growth. Excluding exposures at wavelengths within the resonant feature, longer wavelengths are less effective at reducing the viability of E. coli C1a. This indicates the occurrence of at least two absorption processes.

Absorption↗
PubMed2006

Use of refractometry and colorimetry as field methods to rapidly assess antimalarial drug quality.

The proliferation of counterfeit and poor-quality drugs is a major public health problem; especially in developing countries lacking adequate resources to effectively monitor their prevalence. Simple and affordable field methods provide a practical means of rapidly monitoring drug quality in circumstances where more advanced techniques are not available. Therefore, we have evaluated refractometry, colorimetry and a technique combining both processes as simple and accurate field assays to rapidly test the quality of the commonly available antimalarial drugs; artesunate, chloroquine, quinine, and sulfadoxine. Method bias, sensitivity, specificity and accuracy relative to high-performance liquid chromatographic (HPLC) analysis of drugs collected in the Lao PDR were assessed for each technique. The HPLC method for each drug was evaluated in terms of assay variability and accuracy. The accuracy of the combined method ranged from 0.96 to 1.00 for artesunate tablets, chloroquine injectables, quinine capsules, and sulfadoxine tablets while the accuracy was 0.78 for enterically coated chloroquine tablets. These techniques provide a generally accurate, yet simple and affordable means to assess drug quality in resource-poor settings.

Antimalarials↗
PubMed1995

Determination of anaesthetic agent concentration by refractometry.

Reference refractivity values have been derived for the anaesthetic agents halothane, isoflurane, enflurane, sevoflurane and desflurane which are traceable to national measurement standards. A simple method and equation have been derived for the application of these data to the measurement of agent concentration by refractometry. The main instrumental sources of uncertainty associated with this method are discussed and their respective contributions quantified. Agent concentration can be measured routinely at the +/- 1% level of uncertainty using this approach.

Anesthetics, Inhalation↗
PubMed1983

Refractometry of fungi.

Immersion refractometry is based on matching the refractive index of the medium to that of the cytoplasm of the cell, in which case the latter will have minimum contrast in the phase contrast microscope and will practically disappear. The cytoplasm is then said to be at the match point with the medium. Since the refractive index of a solution is related to its concentration by the formula n-no equal alpha C, the values for the refractive indices can be expressed in terms of the concentration of solids (g/100 ml) which in fact has been done throughout this work. The properties of a suitable immersion medium, its preparation and interpretation of numerical results have been discussed. Refractometric measurements were carried out on fungal hyphae and spores. The solid concentration in hyphae varies according to their age, physiological state and function. In general the highest values are found at sites of physiological activity, such as growth and reproduction, and may range from 11% to 30% of solid concentration. The solid concentration of mature, resting spores was found to be in the range of 45-54%, with the exception of a powdery mildew spores in which the concentration of solids was 30-32%. The germination of spores is preceded by a fall in the concentration of solids, and subsequent swelling before the emergence of the germ tube. When the solid concentration in pre-germinating spore has fallen to about 20-30%, metabolic activity can take place and germination can occur.

Fungi↗
PubMed1985

Comparison of hydrometry, refractometry, osmometry and Ames N-Multistix SG in estimation of urinary concentration.

The four currently available methods used for estimating urinary concentration were assessed. One hundred and fifty urine specimens were collected and 130 were measured. Twenty were excluded as they contained protein and/or glucose. The volume of 41 samples was inadequate for hydrometry. In urines not containing dissolved macromolecules, hydrometry, refractometry and osmolality reliably reflect the degree of concentration. Except at the extremes, the reagent strip on N-Multistix SG is not a reliable indicator of specific gravity. In a children's Hospital, one-third of urine specimens will be of insufficient volume for standard hydrometry.

Child↗
PubMed1982

Photometric immersion refractometry of bacterial spores.

Photometric immersion refractometry was used to determine the average apparent refractive index (n) of five types of dormant Bacillus spores representing a 600-fold range in moist-heat resistance determined as a D100 value. The n of a spore type increased as the molecular size of various immersion solutes decreased. For comparison of the spore types, the n of the entire spore and of the isolated integument was determined by use of bovine serum albumin, which is excluded from permeating into them. The n of the sporoplast (the structures bounded by the outer pericortex membrane) was determined by use of glucose, which was shown to permeate into the spore only as deeply as the pericortex membrane. Among the various spore types, an exponential increase in the heat resistance correlated with the n of the entire spore and of the sporoplast, but not of the isolated perisporoplast integument. Correlation of the n with the solids content of the entire spore provided a method of experimentally obtaining the refractive index increment (dn/dc), which was constant for the various spore types and enables the calculation of solids and water content from an n. Altogether, the results showed that the total water content is distributed unequally within the dormant spore, with less water in the sporoplast than in the perisporoplast integument, and that the sporoplast becomes more refractile and therefore more dehydrated as the heat resistance becomes greater among the various spore types.

Bacillus cereus↗
PubMed1984

Phase separation between nucleoid and cytoplasm in Escherichia coli as defined by immersive refractometry.

The refractive indices of nucleoid and cytoplasm in Escherichia coli were derived theoretically and experimentally. For the theoretical estimates, we made use of the known macromolecular composition of E. coli B/r (G. Churchward and H. Bremer, J. Theor. Biol. 94:651-670, 1982) and of estimates of cell and nucleoid volumes. These were obtained from micrographs of living bacteria made with a confocal scanning light microscope. The theoretical values were calculated, assuming that all DNA occurred in the nucleoid and that all protein and RNA occurred in the cytoplasm. Comparison with experimental refractive index values directly obtained by immersive refractometry showed that, besides its DNA, the nucleoid must contain an additional amount of solids equivalent to 8.6% (wt/vol) protein. With the nucleoid containing 6.8% (wt/vol) DNA and 8.6% (wt/vol) protein and the cytoplasm containing 21% (wt/vol) protein and 4% (wt/vol) RNA, a mass difference is obtained, which accounts for the phase separation observed between the nucleoid and cytoplasm in living cells by phase-contrast microscopy. The decrease in the refractive index of the nucleoid relative to that of the cytoplasm observed upon, for instance, OsO4 fixation was interpreted as being indicative of the loss of protein content in the nucleoid.

Chloramphenicol↗
PubMed2003

Bifocal optical coherenc refractometry of turbid media.

We propose and demonstrate a novel technique, which we term bifocal optical coherence refractometry, for the rapid determination of the refractive index of a turbid medium. The technique is based on the simultaneous creation of two closely spaced confocal gates in a sample. The optical path-length difference between the gates is measured by means of low-coherence interferometry and used to determine the refractive index. We present experimental results for the refractive indices of milk solutions and of human skin in vivo. As the axial scan rate determines the acquisition time, which is potentially of the order of tens of milliseconds, the technique has potential for in vivo refractive-index measurements of turbid biological media under dynamic conditions.

Animals↗
PubMed1999

On-line monitoring of the intravascular volume during haemodialysis by continuous refractometry.

The control of intravascular volume (IVV) by continuous on-line measurement of protein concentration would optimise the patients' specific rate of ultrafiltration. To prove the accuracy of a refractometric device, plasma was continuously drawn by haemofiltration during 10 haemodialysis treatments of male patients. Refractometry reflects highly significant changes in the concentrations of filtrate proteins (r = 0.862, p < 0.001) and blood proteins (rtotal = 0.593, ptotal < 0.001). In vitro, the refractometric device detected a change of protein concentration of 0.041 g/L through a refraction increase of 0.1 mV. The power of discrimination was 0.067% of IVV However, in vivo, the accuracy of IVV refractometric monitoring is reduced by interference factors such as sodium (0. 141 mV/mmol/L), glucose (0.034 mV/mg/dl) and triglycerides (-0.040 mV/mg/dl). Adjustment of the refraction data using sodium and glucose electrodes and plasma filters with a cut-off below the size of chylomicrons is recommended.

Blood Proteins↗
PubMedDate not supplied

Determination of chicken and turkey plasma and serum protein concentrations by refractometry and the biuret method.

Plasma and serum protein concentrations were determined in chickens and turkeys by refractometry (with human and veterinary refractometers) and by the biuret method. Chicken and turkey serum protein values were significantly lower than respective plasma protein values according to both methods. Refractometer readings for both plasma and serum correlated closely with the results of the biuret test (r2 = 0.72 to 0.97). These findings indicate that plasma and serum protein values may be determined accurately in chickens and turkeys with a handheld refractometer.

Analysis of Variance↗
PubMed2004

Evaluation of on-line high-performance size-exclusion chromatography, differential refractometry, and multi-angle laser light scattering analysis for the monitoring of the oligomeric state of human immunodeficiency virus vaccine protein antigen.

Chiron has developed a novel mutant form of the human immunodeficiency virus (HIV) envelop protein, o-gp140, that is currently entering Human Phase 1 clinical trials for testing as a prophylactic HIV vaccine. The o-gp140 protein is oligomeric and the quaternary structure is thought to play an important role in its activity as an antigen. As o-gpl40 proceeds through the clinical trial process and prior to marketing approval, analytical methods that are able to demonstrate manufacturing consistency with respect to degree of oligomerization will need to be developed and validated. On-line high-performance size-exclusion chromatography, differential refractometry, and multi-angle laser light scattering analysis (HPSEC-RI-MALLS), a method commonly used to obtain the molar mass of macromolecules based on the Rayleigh-Gans-Debye approximation, was evaluated for this purpose. The results obtained demonstrated intra- and inter-day precisions to be 0.9 and 3.6% R.S.D., respectively. Accuracy was found to be equal to, or better than, 11% when comparing the known molar masses of test proteins to that of the molar masses determined by the method. Additionally, the method compared favorably to orthogonal native polyacrylamide gel electrophoresis and ultracentrifugation analyses. R-factor analysis was used to demonstrate that HPSEC-RI-MALLS is capable of discriminating compositional differences between o-gpl40 test lots. Based on the data presented, HPSEC-RI-MALLS may be a suitable manufacturing control method.

AIDS Vaccines↗
PubMed2001

Applications of dynamical diffraction under locally plane wave conditions: defects in nearly perfect crystals and X-ray refractometry.

In a previous paper, the concept of a locally plane wave was explained theoretically. In such a configuration, the fringe pattern recorded on the film can be considered as a phase analyser. Here the experimental analysis is presented, showing examples of interesting applications to X-ray refractometry and to the visualization of the strain field around isolated defects.

Journal Article↗
PubMed1973

Immersion refractometry of isolated bacterial cell walls.

Immersion-refractometric and light-scattering measurements were adapted to determinations of average refractive indices and physical compactness of isolated bacterial cell walls. The structures were immersed in solutions containing various concentrations of polymer molecules that cannot penetrate into wall pores, and then an estimate was made of the polymer concentration or the refractive index of the polymer solution in which light scattering was reduced to zero. Because each wall preparation was heterogeneous, the refractive index of the medium for zero light scattering had to be estimated by extrapolation. Refractive indices for walls suspended in bovine serum albumin solutions ranged from 1.348 for walls of the rod form of Arthrobacter crystallopoietes to 1.382 for walls of the teichoic acid deficient, 52A5 strain of Staphylococcus aureus. These indices were used to calculate approximate values for solids content per milliliter, and the calculated values agreed closely with those estimated from a knowledge of dextran-impermeable volumes per gram, dry weight, of the walls. When large molecules such as dextrans or serum albumin were used for immersion refractometry, the refractive indices obtained were for entire walls, including both wall polymers and wall water. When smaller molecules that can penetrate wall pores to various extents were used with Micrococcus lysodeikticus walls, the average, apparent refractive index of the structures increased as the molecular size of probing molecules was decreased. It was possible to obtain an estimate of 1.45 to 1.46 for the refractive index of wall polymers, predominantly peptidoglycans in this case, by extrapolating the curve for refractive index versus molecular radius to a value of 0.2 nm, the approximate radius of a water molecule. This relatively low value for polymer refractive index was interpreted as evidence in favor of the amorphous, elastic model of peptidoglycan structure and against the crystalline, rigid model.

Arthrobacter↗
PubMed2004

Refractometry of microscopic objects with digital holography.

Digital holography has some desirable properties for refractometry of microscopic objects since it gives phase and amplitude information of an object in all depths of focus from one set of exposures. We show that the amplitude part of the image can be used to observe how the Becke lines move between different depths of focus and hence determine whether an object has a higher or a lower index of refraction than its surrounding medium, i.e., the sign of the relief. It is also shown that one single-phase image provides an independent technique to determine the sign of relief between an object and the surrounding medium.

Journal Article↗
PubMed2006

Real-time differential refractometry without interferometry at a sensitivity level of 10(-6).

We present a refractometer based on the principle of total internal reflection that can sensitively record, in real time, the refractive index of fluids over a wide range of refractive indices. The device uses a divergent laser beam and a linear diode array, and has no mechanical or optical moving parts, enabling us to achieve the measurement of a refractive index at a sensitivity level of 10(-6). Our refractometer does not rely on interferometry, thus enabling the device to be compact, portable, and inexpensive. To the best of our knowledge, this is the first time a noninterferometric device that performs real-time differential refractometry with a sensitivity of better than 10(-5) has been demonstrated in the literature. We show that our experimental results agree very well with Fresnel theory. We establish a theoretical limit on the sensitivity of this class of refractometers.

Journal Article↗
PubMed1978

Measurement of neonatal equine immunoglobulins for assessment of colostral immunoglobulin transfer: comparison of single radial immunodiffusion with the zinc sulfate turbidity test, serum electrophoresis, refractometry for total serum protein, and the sodium sulfite precipitation test.

Four procedures for assessment of adequacy of colostral immunoglobulin (Ig) transfer in foals were evaluated. Results of zinc sulfate turbidity test, serum electrophoresis, total serum protein refractometry, and sodium sulfite precipitation test were compared with immunoglobulin G content determined by single radial immunodiffusion. The zinc sulfate turbidity test gave acceptable results for IgG, except that hemolyzed serum samples gave higher than expected values. A correction factor for hemolyzed serum was found to be useful. Serum electrophoresis was a satisfactory method of estimating IgG content. Total serum protein values may not be a valid basis for estimating IgG content, inasmuch as postsuckling total protein values were found to decrease in some foals in which passive transfer of IgG had been adequate. Sodium sulfite precipitation reactions were too unpredictable to be of value for determination of neonatal IgG concentration.

Animals↗
PubMed1998

[Automatic refractometry, reliability of the determination of type and degree of refraction anomalies].

The authors report on their experience with and results of using the AR-1000 autorefractometer made by Nidek, particularly with the pre-school children, school children, and students up to 24 years of age. They stress the significance, as well as speed and accuracy of automatic refractometry findings in large groups of testees (regular checkups) as a valuable indicator in one of numerous necessary tests with the aim to determine and correct visual acuity. There is a significant error reduction in the estimate of dioptries when autorefractometry is done in cycloplegia as compared to autorefractometry without cycloplegia.

Adolescent↗