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Brilliant iridescent colouring in male butter£ies enables long-range conspeci¢c communication and it has
long been accepted that microstructures, rather than pigments, are responsible for this coloration. Few
studies, however, explicitly relate the intra-scale microstructures to overall butter£y visibility, both in
terms of re£ected and transmitted intensities and viewing angles.
Using a focused-laser technique, we investigated the absolute re£ectivity and transmissivity associated
with the single-scale microstructures of two species of Morpho butter£y and the mechanisms behind their
remarkable wide-angle visibility. Measurements indicate that certain Morpho microstructures re£ect up to
75% of the incident blue light over an angle range of greater than 1008 in one plane and 158 in the other.
We show that incorporation of a second layer of more transparent scales, above a layer of highly irides-
cent scales, leads to very strong di¡raction, and we suggest this e¡ect acts to increase further the angle
range over which incident light is re£ected.
Measurements using index-matching techniques yield the complex refractive index of the cuticle mate-
rial comprising the single-scale microstructure to be n (1.56 0.01) + (0.06 0.01)i. This ¢gure is
required for theoretical modelling of such microstructure systems.
Keywords: iridescence; multilayer interference; di¡raction; butter£y; structural colour
Proc. R. Soc. Lond. B (1999) 266, 1403^1411 1403 & 1999 The Royal Society
Received 24 March 1999 Accepted 20 April 1999
1404 P.Vukusic and others Optical characterization of single butter£y scales
Figure 2. Optical microscope images of single scales from M. rhetenor and M. didius. Top surface of the M. rhetenor iridescent ground
scale in (a) re£ection and (b) transmission. (c) Top surface of the M. didius iridescent ground scale in re£ection and (d) in transmission.
Top surface of the M. didius glass scale in (e) re£ection and ( f ) transmission. (Note that the camera has compensated for low light
levels and the blue colour of the scale shown in (e) is arti¢cially enhanced.) Scale bar in (a) for (a,b,c,d,e, f ), 100 mm.
The characteristic blue iridescence exists only in re£ection responsible for the iridescent colour. This structure is one
from the top surface of each scale. The blue of the of the two main variants of multilayer structure found in
M. didius glass scale (¢gure 2e, f ), which in comparison Lepidoptera that display iridescence. The other, found for
with the blue of the ground scales is of relatively low example in iridescent scales of Urania moths (Lippert &
intensity, appears equally intense due to the contrast Gentil 1959), is formed from multilayers that exist within
compensation of the camera. the substrate of the scale itself.
It is interesting to compare the transmitted colours of The spatial patterns of re£ection and transmission
each scale. The ground scales of both species are dark from single iridescent ground scales of both butter£ies
brown. This colouring derives both from the optical and from a glass scale from M. didius are shown in ¢gure
absorption associated with pigment and the strong blue 4a,b. For the ground scales in air, with the beam incident
re£ection from the structure on the top surface. The normal to the scale's top surface and the scale oriented so
M. rhetenor ground scale appears darker than that of that the ridges run vertically, a broad re£ection of speckle
M. didius, due either to a higher concentration of melanin is recorded, spanning over 1008 in the horizontal plane
or to the presence of a more absorbing form of melanin and ca. 158 in the vertical plane.
(Nijhout 1991). The M. didius glass scale appears highly Re£ection from the M. rhetenor ground scale appears
transparent, indicating the absence of melanin. divided into two distinct lobes around the normal to the
In ¢gure 3, SEM images of the Morpho scales, and more scale surface (shown in ¢gure 4a, for a 488 nm incident
particularly TEM images of their cross-sections, reveal wavelength). TEM images of this scale indicate a distribu-
the periodic upper-surface ridges containing multilayers tion of tilts of the multilayer upper and lower surfaces,
(a)
120 60
1.6
refln
in IPA
scale
180 0
1.2
transn
intensity (a.u.)
240 300
0.8 120 60
refln
scale
in air
0.4 180 0
transn
240 300
0.0
scale
in air
scale
in IPA
reflection transmission
0 60 120 180 240 300 360
polar angle around single Morpho rhetenor scale
0.8 (b)
120 60
refln
ground
scale
0.6 180 0
transn
intensity (a.u.)
240 300
0.4
120 60
refln scale
glass
180 0
0.2
transn
240 300
0.0
ground
scale
glass
scale
reflection transmission
Figure 4. (a) Linear and polar plots of experimental data (using 488 nm TM ö polarized incident light), showing 3608 angle
scans around a single M. rhetenor iridescent ground scale in air and in IPA. (b) Linear and polar plots of experimental data (using
488 nm TM ö polarized incident light), showing 3608 angle scans in air around a single M. didius iridescent ground scale and
iridescent glass scale. (NB (i) A 908 polar angle corresponds to normal incidence on the scale. (ii) Polarizations are de¢ned as:
TM, electric ¢eld parallel to ridges; TE, electric ¢eld perpendicular to ridges.
75
reflection (%)
75 50
absolute
25
absolute reflection (%)
0
300 400 500 600
50
20
transmission (%)
15
absolute
10
25
5
0
450 500 550 600
wavelength (nm)
0
450 500 550 600 650
wavelength (nm)
Figure 5. Normal incidence wavelength-dependent re£ectivity for M. rhetenor and M. didius single iridescent ground scales (dotted
lines in these two graphs represent guides to the eye). (Open circles and crosses represent TM polarization data for M. rhetenor and
M. didius, respectively. Filled circles and diagonal crosses represent TE polarization data for M. rhetenor and M. didius, respectively.)
The top inset graph shows these single-scale data and also spectrometer data for large-area wing sections of both butter£ies extending
into the UV. The solid line represents TM polarization data for M. rhetenor and the dashed line represents TM polarization data for
M. didius. TE polarized data are not presented here. For comparison of the absolute single-scale data and the large-area spectrometer
data, the latter are scaled by a factor of 1.15 to normalize the e¡ect of spectrometer optics and non-single-scale experimentation on
measured intensities. The bottom insert graph shows transmissivity for both single iridescent ground scales. (Polarizations are de¢ned
as: TM, electric ¢eld parallel to ridges; TE, electric ¢eld perpendicular to ridges.)
the light collected in re£ection from, and transmission Polarization di¡erences do not signi¢cantly a¡ect
through, each scale) are shown in ¢gure 5. Spectrometer re£ected light intensities for the M. didius ground scale,
data, taken for large-area wing sections of each butter£y, but there is a clear di¡erence in the M. rhetenor scale. This
are presented in the top inset graph in ¢gure 5 (overlaid possibly indicates a degree of polarization sensitivity in
on the single-scale data). the photoreceptors of the M. rhetenor species, already
The re£ectivity data con¢rm what is discernible to found to exist in honey-bees (von Frisch 1965). Detailed
human vision; namely, the upper-wing surface of work on the visual system of this species is necessary to
M. rhetenor is a deeper and more intense blue than that of con¢rm this.
M. didius. The peak re£ectivity of ca. 40% (measured The spectrometer-generated re£ectivity results (see top
using the single-scale technique) at ca. 475 nm for inset graph in ¢gure 5), indicate the form of the wing
M. didius contrasts with the peak re£ectivity of ca. 70% at re£ectivity at visible and UV wavelengths. These data are
450 nm for M. rhetenor. This di¡erence in absolute re£ec- subject to the inconsistencies of large-area measurement
tivity is caused by the greater number of layers within the described earlier, but they nonetheless indicate the extent
M. rhetenor ridging (¢gure 3b,d), and is predicted by multi- of re£ection in the UV. For both species of Morpho, this
layer theory (Huxley 1968). It is worth stating that this UV component of wing re£ection is clearly signi¢cant
theory predicts near-100% peak-wavelength re£ection in and must contribute substantially to the perception of
a system with upwards of ¢ve quarter-wave multilayers. wing hue by UV-sensitive eyes.
However, the Morpho multilayers are not structured on an Absolute re£ectivity measurements for the M. didius
exact quarter-wave basis and therefore the re£ectivity single glass scale in air, which are not presented here,
falls below 100%. Further reduction in peak re£ected show a peak of 5^10% at blue wavelengths, decreasing to
intensity is produced by the discrete nature of the multi- ca. 3% at other wavelengths.
layering and through the presence of optical absorption in The bottom insert graph in ¢gure 5 shows that absolute
the scales' structure. transmission values through both types of ground scale in
Experimental evidence supports this hypothesis, indi- e¡ects, both of which enhance the angular spread of light
cating that not only are males attracted to visual stimuli re£ected from the wing. First, the presence of multilayer
resembling females, but that they can be repelled from surfaces that show a distribution of tilts with respect to
long distances by visual stimuli resembling other males the scale substrate creates a very large spread in angle
(Obara 1970; Stride 1958; Rutowski 1978, 1981; Silberglied over which the characteristic blue light is re£ected.
& Taylor 1978). Brilliant wing coloration appears to have Similar multilayer tilting, increasing the angular spread
the capacity to serve as an agonistic device that is used of re£ected light, has been found in the re£ectors of the
for threat (Hingston 1933) and for intimidation of rivals crab Ovalipes (Parker 1998).
for prime locations (Shields 1968). It has developed in In addition, the existence of a second layer of periodi-
male butter£ies to promote their visibility while in £ight cally ridged but highly transparent glass scales above the
or while resting with open wings. layer of highly iridescent ground scales causes strong
Although some butter£y pigmentary colours appear di¡raction. This di¡raction does not appear to serve the
bright and highly visible to human eyes, their absolute purpose of optical dispersion, but acts to spread further
visibility is substantially inferior to those wing colours the angle over which incident light is re£ected. A purely
that are generated through optical interference. Measure- spectral re£ection from a mirror-like surface would
ments in this study indicate that over 70% of incident severely restrict the solid angle from which the wing can
blue light is re£ected from the intra-scale structure of be observed. Long-range communication would in this
M. rhetenor, far above what can be achieved through respect be limited. In the case of the Morpho butter£ies
pigmentary coloration alone. Selection pressures on these studied here, incorporation into the optical system of
species have caused the evolution of large numbers of either a multilayer surface-tilt distribution, or di¡raction-
multilayers that are appropriately dimensioned for produ- assisted angle broadening using glass scales, or both,
cing this very signi¢cant re£ectivity. Such structural enhances their overall angular visibility.
coloration also enables change of hue with wing orienta-
tion, narrow spectral purity, the possibility of a strong We thank Gavin Wakely at the E.M. Unit in the School of
component of UV re£ection, and distinct polarization Biological Sciences of Exeter University for expert technical
e¡ects that are also not attainable with pigments. assistance, and David Bolton at the Royal Albert Museum,
Sheer brightness is very likely to serve its main purpose Exeter, UK, for access to specimens. This work receives funding
in long-range communication. In terms of the spectral from the BBSRC, the DERA (Farnborough) and Exeter
response of human eyes, Morpho can easily be seen from University.
low-£ying aircraft (Silberglied 1984), and Bates reported
visibility from `a quarter of a mile o¡ ' (Bates 1864).
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