1. Introduction
Color vision disorders occur among the symptoms of numerous diseases of the human eye, including glaucoma [
1,
2], cataract [
3,
4], optic neuritis [
5], and certain systemic diseases, such as diabetes mellitus [
6,
7]. Changes in color perception may affect the areas of red, green, and blue related to three types of cones (L, M, and S) in the retina, which are responsible for perception of three primary colors. Although color vision disorders are observed in the above diseases, they are not used to assess their severity. Such approach may be explained by several reasons. The first and the most important one is the scarcity of scientific research describing color vision deficits in particular diseases in a quantitative manner, the second is the scarcity of standardized diagnostic methods that would quantify color vision deficits, in particular in the green/blue range. Another issue is the inter-subject variability of color perception, which makes defining norms for healthy individuals ambiguous in certain cases.
Such variability might be related to:
- (a)
variability of primary structure of receptor proteins and minor peak shifts or other changes in their absorption spectra [
8],
- (b)
variable number of cones of each type in the retina and the macula [
9],
- (c)
differences in the structure or functioning of the retina’s neuronal network, which codes information concerning the activity of each cone type and transfers the color information to the central nervous system [
10],
- (d)
differences in color perception at the brain cortex level.
Particularly, aspects (c) and (d) above have not been well-researched so far in the literature.
Despite these limitations, developing a methodology for quantifying color perception and analyzing changes in color perception in individual diseases is an important direction of research. In the studies carried out to date in our laboratory, the researchers have assessed the anomaloscopic test, which enables quantitative assessment of color perception in the red/green (Rayleigh test) and green/blue (Moreland test) [
11] axes. The Rayleigh test is well standardized and the results are comparable. This test is primarily used to quantify deficiencies of red–green perception. In contrast, the Moreland test has been poorly studied and is practically not used for diagnostic purposes in clinical practice. The Rayleigh test is based on a comparison of a subjective impression of the color yellow (produced by mixing green and red) with a standardized yellow color. As a result of the measurement procedure, border values of light intensity ratio are defined for the red and the green diodes (R
1 and R
2). The R value can range between 0 (100% green diode) and 73 (100% red diode) anomaloscope units [
12]. The diodes allow the subject to perceive the test color as being identical to the reference color. An additional parameter is the brightness of the yellow reference color at the color adjustment borders (Y
1, Y
2). Similarly, the Moreland test is based on comparing the subjective impression of the cyan in the test field (resulting from mixing the light emitted by green and blue diodes) with the standard cyan diode. The resulting measurement values are the border values of intensity for the blue and the green diode, which give make the subject perceive the test color as being identical to the standard cyan color (M
1 and M
2). Another test outcome is the brightness of the reference cyan color at the time of shade equalization (C
1, C
2). The M value can range between 0 (100% green diode) and 100 (100% blue diode) anomaloscope units.
Recently, Zabel et al. [
11] presented the results of anomaloscopic measurements of Polish subjects aged below 45 years without color vision disorders, as determined by the HRR method. The subjects were free from eye diseases that might affect changes in color perception. The Rayleigh test results obtained by the research team, as compared to the standards recommended by the anomaloscope manufacturer, were slightly shifted towards protanomaly. The recommended standard according to the anomaloscope user’s manual is 34–46 units [
12], while the results obtained in the study (±2 std) were 35.4–51.3 for females and 36.8–54.8 for males. Additionally, it was observed that the distribution of the R
2 parameter was characterized by two maxima with a minimum between the peaks located around the value of 50. Values above 50 were observed mainly in male subjects. Based on the above observations, it was suggested that approximately 20% of males should be classified as a subtle protanomaly group. The result may be related to the fact that opsin (protein) is encoded on the X chromosome and men have only one copy of this gene. However, such an assumption poses a problem in determining a norm for the male population. The norms suggested above, however, include 2 std around the mean (95% of the population) and therefore the majority of the subtle protanomaly peak is included in the norm.
The results obtained for the Moreland test differed significantly from the norm proposed by the anomaloscope manufacturer. It was observed that the fitting range was significantly wider (MC = M2 − M1) and considerable inter-subject variability for the border parameters M1 and M2 was also noticeable. In particular, such variability was observed in case of the M1 parameter. For this reason, the Moreland test in its current version seems to have little value for quantification of color vision disorders in the green/blue axis.
In the majority of subjects, the impression of blue was dominant in a wide range of the proportion of green and blue diode intensities. Thus, one of the possible explanations of the large variability of the Moreland test results obtained in this study was too high intensity of the blue diode, which dominated over the green. In order to investigate this hypothesis in the present work, the Rayleigh and Moreland test were administered using additional lenses which partially filtered blue light. Among the available filters, the researchers selected three: Blue Control (F-BC) by Hoya, Med. 1 filter (F-Med1) manufactured by JZO and F-450 by Eschenbach. The results obtained using each of the three filters were compared against test results without any filter (F-0). It was expected that reducing the intensity of the blue diode should result in a reduction of the extremely high value of the M2 parameter and a reduction of the variability of M1 values. Consequently, a reduced width of the matching range (MC) could contribute to a greater diagnostic value of the test with the use of a specific filter.
Anomaloscopic examination enables the assessment of three aspects of color vision:
The first aspect is the assessment of the relative subjective brightness of the tested primary colors red, green, and blue, which correspond to the maxima of absorption spectra of subsequent cones L, M, and S.
The second one is the ability to perceive changes in the different shades and tones, i.e., neighboring RGB colors may be perceived either as the same or slightly different in quality.
The third aspect is quite similar to the first one but, nevertheless, it seems reasonable to distinguish between them. It is the general ability to perceive the given color being understood as the sensitivity of the given retinal cones to the given wavelength.
The assessment of relative subjective brightness may be assessed based on the Y = fn(R) relation in the Rayleigh test and the C = fn(M) relation in the Moreland test, where Y is the luminance of the yellow reference diode as a function of R (the proportion of red to green), and C is the luminance of the cyan reference diode as a function of M (the proportion of blue to green). In the case of the Moreland test, the sensitivity to blue light is generally lower than sensitivity to green light and therefore the slope (aM = (C2 − C1)/(M2 − M1) is positive and approximately equal to 1. For example, if the evaluated individual perceives 436 nm blue light as relatively darker than it is perceived by others, then the aM slope for the C = fn(M) relation is higher. As the M setting is increased (i.e., more green is added and blue is reduced), the perceived brightness increases more rapidly. In the Rayleigh test, red is perceived as a relatively darker color than green, therefore, aR is negative. If a given subject perceives red as being darker (in comparison to other individuals’ perception), then, as the proportion of red is increased, the perceived brightness decreases at a higher rate and the aR ratio becomes more negative.
The assessment of the ability to perceive subjective changes in the color tones between blue and green (sea color/cyan/aquamarine/turquoise/olive) may be defined as the width of the matching range (MW = M2 − M1, RW = R2 − R1). The more narrow the color matching range, the more efficiently the adjacent shades and tones are perceived.
As for the third aspect, i.e., the overall ability to perceive colors, a reduced ability to perceive a particular primary color does not necessarily mean that the color will be perceived as darker. For example, a reduced ability to perceive red will be manifested as a shift of the parameter to the right. It means that a more red diode must be added for the resulting color (i.e., red mixed with green) to be perceived as identical to the reference yellow.
However, this does not necessarily mean that exemplary red will be perceived by the subject as darker (a higher slope in the Y = fn(R) relation). The overall ability to perceive a particular color will be manifested mainly when it is mixed with one of other primary colors. The differentiation between the perceived color brightness and the ability to perceive a given color has not been well researched to date and has not been discussed in the literature. In general, it is more complex to analyze than the two previously mentioned aspects.
It should be emphasized that if anomaloscope is to be used to detect subtle changes in color perception (due to disease processes) rather than to detect major deficits (related to genetic defects), the three aspects must be understood and analyzed separately. In addition, a perception disorder related to a particular disease may affect only one of the above aspects. So far, anomaloscopic examinations have been mainly based on the Rayleigh test administered in order to assess vision of the red color, i.e., the need to increase the amount of red light during the test in order to achieve an impression of yellow in the test field that matches the color in the reference field. It is manifested by a shift of R2 and RC to the right and widening of the matching range (RW). The Y parameter of the Y = fn(R) function has not been used to date to assess color vision disorders resulting from particular disease processes.
In this article, apart from the assessment of blue light filters usability, an analytical proposal for the application of the above parameters is described. The analyzed parameters may be obtained from anomaloscopic examinations using Rayleigh and Moreland tests.
4. Discussion
The data presented in
Table 5 and in
Figure 5 indicates that the results of the Moreland test are significantly correlated mainly with the values of R
W and, to a lesser extent, with R
1 and R
2. R
W is related to the ability to distinguish color tones in the lemon-yellow-orange axis. It is noticeable that a good ability to differentiate color tones in the Rayleigh test also allowed the subjects to better differentiate between color tones in the green–blue axis. In the case of F-0 tests, such effect was noticeable primarily as an increase in M
1 values i.e., the parameter, which shows significant inter-subject variability. M
1 was also correlated to R
1, which leads to the conclusion that the correlation mainly depends on the quality of green perception. Efficient perception of green (i.e., high R
1 value) causes statistically higher values of M
1 and more narrow matching ranges (M
C) in the Moreland test.
When blue filters were applied, the relation was somewhat different. The correlation between RW and other Moreland test parameters was still observable while the relationship with R1/R2 was altered. The correlation to R1 diminished in case of the F-BC filter, a strong correlation to R2 occurred for the F-Med1 filter, while the correlations both to R1 and R2 were no longer noticeable in case of the F-450 filter.
It should be noted that, in case of the F-Med1 filter, the correlations reached values between 0.6 and 0.8, which makes the results of both tests significantly aligned.
Since R2 is mainly related to the quality of red perception, it is reasonable to conclude that a reduction of blue light and, to a lesser extent, the amount of green light, when the F-Med1 filter is applied, increases and emphasizes the role of L cones and the L-M information in the perception of blue and green light as well as color tones in the green-cyan-blue axis. Proficiency in differentiating color tones in the lemon-yellow–orange axis increases the competence in differentiating between the color tones in the blue-cyan-green axis.
In case of F-Med1, there is also a clear negative correlation of aM to R2 and to RC. This means that low values of R2 and RC (i.e., better red perception) is associated with higher aM values (i.e., blue is relatively darker or green is brighter). It can be carefully concluded that higher individual number of red cones in the retina is correlated with lower number of blue cones rather than with the green ones and blue light reduction magnifies this effect.
It is arguable that a decrease in luminance of the blue diode in the Moreland test enhances the correlations between the tests in terms of green perception which is common for both test types. Such result is another indication that the intensity of light emitted by the blue diode in the Moreland test is too high and reducing it by 15–40% percent may be beneficial in the context of detecting certain abnormalities. This may be applicable especially in case of decreased green perception ability.
An important observation was also that the correlations between RC and MC were low while the correlations between R1, R2, M1, and M2 were high, which indicates that the limit parameters (R1, R2, M1, and M2) should be considered independently from one another and that calculation of mean values makes the related information less convincing. RC and MC are significant in cases of more pronounced color vision deficits but they are less meaningful when more subtle differences are considered.
The results discussed above indicate that there is a significant and unused potential of anomaloscopic examination in terms of quantitative analysis of subtle inter-subject differences in terms of color perception. The same is also true for this examination in the context of quantitative assessment of deterioration in the perception of certain colors in the course of various disease processes affecting the visual and nervous systems. The application of blue light filters presented herein suggests that there is a significant potential for further analyses and new evaluation parameters in cases of color vison disorders due to an underlying disease process.
One of the main difficulties, which hinders the acquisition of valuable results related to disease severity in the context of color vision, is inter-subject variability. The application of filters, including those with different parameters such as those reducing red or green light, may provide additional parameters which may be less related to the inter-subject variability and may serve as better indicators of the characteristics of certain disease processes. Further analyses may also shed some light on the theory of opposing colors, the characteristics of color coding in the retina as well as color perception by the central nervous system.
It also seems justified to modify the Moreland test so that the difference between the wavelengths of the green diode (490 nm) and the cyan reference diode (480 nm) becomes more significant. Thus, the color matching area in the Moreland test could be narrowed down, which may facilitate more precise analysis. However, this shall require redesigning of the anomaloscope and a revision of the reference ranges for a new set of diodes used in the green–blue axis test. It might also be interesting to design a test able to compare blue directly to red by means of comparison of the red–blue color mix in the test field with a reference violet diode. Valuable results may also be expected from the application of a filter characterized by significantly different transmittance for 480 nm and 490 nm wavelengths, as this would allow test subjects to better differentiate between the green and cyan diodes (with the design of anomaloscope remaining as is). The filter suggested here could possibly have transmittance levels of 15–40% for 436 nm, 50–80% for 480 nm, and 95–100% for 490 nm. Application of such filter might significantly reduce the excessive width of the matching range (MC) in the Moreland test and could facilitate more efficient diagnosis of blue color vision deficits.