A New Design of Flicker Photometer for Laboratory Colored Light Photometry

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Light as a true visual quantity : principles of measurement

This Technical Report summarizes visual photometric measurement methods which can provide visually meaningful assessments of light. They can be more complicated than the simple use of of a V(?)-corrected physical photometer, and in addition require some understanding of the visual system and how it works. Their advantage is that the assessment of light bears a logical relationship to the human perception of light. For photopic vision and luminances larger than several cd/m2, ordinary physical photometers corrected to V(?) give visually accurate measures for small, centrally fixed, broad-band lights. For other applications, a different luminous efficiency function should be employed. In order to utilize the appropriate function, one must either measure the spectral distribution of radiant power directly or correct the V(?) response of the photometer to the appropriate luminous efficiency. An alternative solution is to calculate mathematical formulas specifically developed for this purpose. This method is potentially the most useful since different formulas can be developed for different applications (for example, two degree or ten degree fields). It is based on established CIE data, and no additional measures need to be developed.For scotopic vision, an assessment of radiant power is made with respect to the scotopic luminous efficiency function V'(?) by means of an appropriately corrected physical photometer, by radiance measurement, or by visual photometry. In mesopic photometry, the photopic and scotopic contributions of the light must be assessed. An estimate can be obtained by combining the photopic and scotopic luminances non-linearly. A more precise measure can be obtained by using three or, still better, four quantities based on X10, Y10, Z10, and V'(?)

Delayed cone-opponent signals in the luminance pathway

Cone signals in the luminance or achromatic pathway were investigated by measuring how the perceptual timing of M- or L-cone-detected flicker depended on temporal frequency and chromatic adaptation. Relative timings were measured, as a function of temporal frequency, by superimposing M- or L-cone-isolating flicker on "equichromatic" flicker (flicker of the same wavelength as the background) and asking observers to vary contrast and phase to cancel the perception of flicker. Measurements were made in four observers on up to 35 different backgrounds varying in wavelength and radiance. Observers showed substantial perceptual delays or advances of L- and M-cone flicker that varied systematically with cone class, background wavelength, and radiance. Delays were largest for M-cone-isolating flicker. Although complex, the results can be characterised by a surprisingly simple model in which the representations of L- and M-cone flicker are comprised not only of a fast copy of the flicker signal, but also of a slow copy that is delayed by roughly 30 ms and varies in strength and sign with both background wavelength and radiance. The delays, which are too large to be due to selective cone adaptation by the chromatic backgrounds, must arise postreceptorally. Clear evidence for the slow signals can also be found in physiological measurements of horizontal and magnocellular ganglion cells, thus placing the origin of the slow signals in the retina-most likely in an extended horizontal cell network. Luminance-equated stimuli chosen to isolate chromatic channels may inadvertently generate slow signals in the luminance channel

GENDER AND OVARIAN HORMONE EFFECTS ON THE RELATIVE CONTRIBUTION OF CHROMATICITY TO BRIGHTNESS

PURPOSE: The chromatic contribution to brightness perception was compared in males and females. METHODS: Direct brightness matching (DBM) and heterochromatic flicker photometry (HFP) were used to measure relative luminous efficiency, and DBM/HFP ratios were predicted to be higher for females than males on repeated measures and for each primary color. No gender differences were predicted in DBM or HFP measures. Within-females effects of estradiol (E2) and progesterone (PG) levels, contraceptive use, and menstrual cycle phase were also investigated. It was expected that E2 would directly predict DBM/HFP ratios and that PG would antagonize that relationship. Based on that prediction, DBM/HFP ratios would be at a maximum during the ovulatory phase, intermediate during the menstrual phase and minimum during the luteal phase. No effects were predicted for DBM or HFP measures. RESULTS: DBM/HFP ratios were significantly higher for female subjects than male subjects. Contraceptive use had no effect on overall DBM/HFP ratios. There were limited effects of hormone levels, menstrual cycle phase and contraceptive use on DBM/HFP ratios and on DBM and HFP measures analyzed separately. However, hormone effects on DBM/HFP ratios and DBM measures at 650 nm agree with previous findings involving long-wavelength sensitive (L-) cone mechanisms. CONCLUSIONS: The present findings supporting a female advantage in chromatic contribution to brightness are robust. The significant results are discussed in the context of previous findings suggesting organizational and short-term effects of ovarian steroid hormones

Multiplexed Photometry And Fluorimetry Using Multiple Frequency Channels

ABSTRACT MULTIPLEXED PHOTOMETRY AND FLUORIMETRY USING MULTIPLE FREQUENCY CHANNELS by KHALED M. DADESH August 2013 Advisor: Dr. Amar Basu Major: Electrical Engineering Degree: Doctor of Philosophy Multispectral photometry and fluorimetry are useful for quantifying and distinguishing samples during flow injection analysis, flow cytometry, and ratiometric absorbance measurement. However, multispectral detectors, including spectrometers, typically require arrayed or multiple light detectors, optical components, and path alignments, all of which increases the size and cost of the detection system. Several previous efforts have attempted the use of time division multiplexing or frequency division multiplexing (FDM) techniques to minimize both size and cost of multispectral photometry equipment by using only a single light detector. Although many of these designs achieved low cost, they generally operated at \u3c50 KHz, which limited the detection speed of the overall system. An alternative frequency multiplexing design operated at 3MHz; however, it required electro optical modulators [50], which are too expensive and bulky for portable applications. In contrast to both approaches, the objective of this research is to use frequency division multiplexing to perform multispectral photometry and fluorimetry while achieving both low cost and high frequency operation (up to 100 MHz). The multiplexing is performed electronically using low cost optoelectronic sources, a single light detector, and a single high-throughput interrogation window. It enables us to perform multi-parameter biological analysis at lower costs and less complexity. Multiple monochromatic light sources, each with a unique wavelength, are electronically modulated at distinct frequencies, and their combined light emission is directed to the sample detection cell. The light transmitted by the sample (absorbance mode) or emitted by the sample (fluorescence mode) is directed to a single light detector. The received light is then converted to a voltage signal and demodulated into the frequency channels using phase-sensitive electronics. Each recovered channel therefore provides either absorbance or fluorescence at its respective optical wavelength. The system is designed to operate at high speed in order to be used in high throughput detectors such as flow cytometers. As a proof of concept, we apply the FDM technique in two detection systems: 1) a three-color absorbance photometry detector and 2) a two-color laser induced fluorescence (LIF) detector. In the first system, three LEDs are operated with 150 KHz, 200 KHz, and 250 KHz modulation frequencies, and the system achieves a 1 ms measurement time constant at an overall component cost \u3c$10. We perform absorbance photometry of four different organic dyes in flow injected solutions and in discrete droplet microreactors with throughputs in the 10\u27s of samples per second. In both cases, the system is able to simultaneously discriminate between them [13]. In the LIF system, first two laser diodes operated at 1 KHz and 1.5 KHz, respectively, are used to excite fluorophores at the respective frequencies. This system is able to distinguish low speed (1 drop/sec) water-in-oil droplets containing fluorescein or rhodamine-6G generated in a microfluidic junction. Second two laser diodes operated at 25MHz and 40 MHz, respectively, are controlled using a developed high frequency FDM system to excite Fluorescein and Alexa 680 dyes at the respective frequencies. Because of the high frequency operation, this system is able to distinguish alternating high speed (300 drops/sec) droplets containing the two fluorescent dyes. In both case, the developed In previous experiments we use an inverted fluorescence microscope with a specific optical cube to excite dyes and collect fluorescence signals. These two FDM-LIF systems identify the different fluorophores based on their excitation frequency rather than their emission band, giving it a unique ability to distinguish fluorophores with overlapping emission spectra. However, overlapping excitation spectra is a problem in the FDM-LIF system, and any assay has to be prepared using fluorophores with minimal excitation overlap. Therefore, fluorophores with sharp excitation lines such as lanthanide ions are the best candidate material in use with FDM-LIF system. The system uses high frequency (100 MHz) modulation which enables multiplexed time constants on the order of 1 µs. Achieving this high bandwidth allow us to apply the system towards high throughput analysis such as cell cytometry, where it could substantially reduce cost and size of the system. Therefore, the FDM-LIF system is installed in an old BD bioscience cytometer, which is available in the cell cytometry laboratory in Karmanos Cancer Research Center (KCRC) located at Detroit Medical Center (DMC). A biological assay containing Alexa Fluor® 680 Goat Anti-Mouse IgG (H+L) and Alexa Fluor® 430 Goat Anti-Mouse IgG (H+L) with BDTM CompBead Anti-mouse Ig, κ beads is tested using the FDM-LIF system. The system is capable to count the two different antigens simultaneously, which gives the possibility of incorporating this system in cytometers. This technology promises to reduce cost and complexity of future cytometers

Evaluating the Talbot-Plateau Law

The Talbot-Plateau law asserts that when the flux (light energy) of a flicker-fused stimulus equals the flux of a steady stimulus, they will appear equal in brightness. To be perceived as flicker-fused, the frequency of the flash sequence must be high enough that no flicker is perceived, i.e., it appears to be a steady stimulus. Generally, this law has been accepted as being true across all brightness levels, and across all combinations of flash duration and frequency that generate the matching flux level. Two experiments that were conducted to test the law found significant departures from its predictions, but these were small relative to the large range of flash intensities that were tested.Comment: 34 pages, five figure

A Notion or a Measure: The Quantification of Light to 1939

This study, presenting a history of the measurement of light intensity from its first hesitant emergence to its gradual definition as a scientific subject, explores two major themes. The first concerns the adoption by the evolving physics and engineering communities of quantitative measures of light intensity around the turn of the twentieth century. The mathematisation of light measurement was a contentious process that hinged on finding an acceptable relationship between the mutable response of the human eye and the more easily stabilised, but less encompassing, techniques of physical measurement. A second theme is the exploration of light measurement as an example of ‘peripheral science’. Among the characteristics of such a science, I identify the lack of a coherent research tradition and the persistent partitioning of the subject between disparate groups of practitioners. Light measurement straddled the conventional categories of ‘science’ and ‘technology’, and was influenced by such distinct factors as utilitarian requirements, technological innovation, human perception and bureaucratisation. Peripheral fields such as this, which may be typical of much of modern science and technology, have hitherto received little attention from historians. These themes are pursued with reference to the social and technological factors which were combined inextricably in the development of the subject. The intensity of light gained only sporadic attention until the late nineteenth century. Measured for the utilitarian needs of the gas lighting industry from the second half of the century, light intensity was appropriated by members of the nascent electric lighting industry, too, in their search for a standard of illumination. By the turn of the century the ‘illuminating engineering movement’ was becoming an organised, if eclectic, community which promoted research into and standards for the measurement of light intensity. The twentieth-century development of the subject was moulded by organisation and institutionalisation. Between 1900 and 1920, the new national and industrial laboratories in Britain, America and Germany were crucial in stabilising the subject. In the inter-war period, committees and international commissions sought to standardise light measurement and to promote research. Such government- and industry-supported delegations, rather than academic institutions, were primarily responsible for the ‘construction’ of the subject. Practitioners increasingly came to interpret the three topics of photometry (visible light measurement), colorimetry (the measurement of colour) and radiometry (the measurement of invisible radiations) as aspects of a broader study, and enthusiastically applied them to industrial and scientific problems. From the 1920s, the long-established visual methods of observation were increasingly replaced by physical means of light measurement, a process initially contingent on scientific fashion more than demonstrated superiority. New photoelectric techniques for measuring light intensity engendered new commercial instruments, a trend which accelerated in the following decade when photometric measurement was applied with limited success to a range of industrial problems. Seeds sowed in the 1920s – namely commercialisation and industrial application, the transition from visual to ‘physical’ methods, and the search for fundamental limitations in light measurement – gave the subject substantially the form it was to retain over the next half-century

Visual performance in the mesopic range

The aims of this work were to assess the effects of different stimulus parameters on chromatic sensitivity, with particular emphasis on the effect of retinal illumination, and to investigate aspects of suprathreshold visual performance under mesopic conditions. All investigations were performed using visual psychophysical techniques. Chromatic thresholds were obtained using dynamic luminance contrast noise to isolate responses to colour signals. The effects of stimulus size and spatial distribution were examined in normal trichromats, dichromats and subjects with acquired colour vision deficiency. The chromatic sensitivity of normal trichromats was investigated with reduction in light level. Measurements were also performed to assess the possible involvement of rods in chromatic processing at threshold. Suprathreshold performance in the mesopic range was assessed in terms of the relative contributions of colour and luminance contrast to a measure of stimulus conspicuity and to visual search time. The conspicuity of a stimulus defined by colour and luminance contrast was defined as the value of achromatic contrast of a similar stimulus with an equal perceived conspicuity. An empirical model was developed from an extensive data set of conspicuity matches, to enable prediction of conspicuity for a wide range of coloured stimuli. Visual search performance for both achromatic and coloured stimuli was investigated under mesopic conditions, and results for the coloured stimuli were compared to the measure of stimulus conspiculty combined with an achromatic search time calibration. The results revealed that chromatic sensitivity is dependent on stimulus size, spatial distribution, eccentricity of presentation and level of illumination. These factors are suggested to reflect changes in cone performance and the relative cone contributions to the postreceptoral chromatic channels. Chromatic sensitivity was found to be independent of rod activity in the mesopic range, suggesting separate processing of rod signals and threshold colour signals under mesopic conditions. Measurements of stimulus conspicuity under mesopic conditions revealed individual variations in response to both luminance contrast and chromatic signals indicative of individual differences in gain control of postreceptoral mechanisms. Conspicuity was successfully modelled as a function of photopic contrast, scotopic contrast, chromatic difference to the background and the level of illumination. The nonlinear relationship between search time and luminance contrast was found to change with reduction in light level, reflecting increased contrast thresholds and diminishing effectiveness of unit physical contrast. Mesopic visual search was also found to depend on the photopic contrast, scotopic contrast and chromatic content of the stimulus, but with an apparent greater emphasis on scotopic contrast and reduced emphasis on colour compared to the measure of stimulus conspicuity. Conspicuity was successfully used to predict visual search times, and was found to be an improved indicator of search performance than either photopic or scotopic contrast