Research Article |
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Corresponding author: Gunnar Brehm ( gunnar.brehm@uni-jena.de ) Academic editor: Maria Heikkilä
© 2025 Gunnar Brehm.
This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Citation:
Brehm G (2025) Light in standardised insect photography and description of lighting devices, including the UV range. Nota Lepidopterologica 48: 145-164. https://doi.org/10.3897/nl.48.145444
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Digital photography has become indispensable in many areas of biology and plays an essential role in digitization in museums and for the analysis of traits of organisms. The aspect of lighting quality has received surprisingly little attention so far, although good colour rendering is an important criterion. This paper provides an overview of the light quality of various lighting systems in three natural history museums. Light quality is usually expressed by the Colour Rendering index. Here, the relatively strict CIE (International Commission on Illumination) Re index, based on the matching of 16 colours, is used. It presents three self-developed lighting devices with different designs: a light cylinder (also suitable for UV photography of individual objects), a light box for photographing entire insect boxes and a light hemisphere for photographing specimens with reflective surfaces. No light source measured in the natural history museums in London and Berlin achieves an Re index value of >90. Compared to daylight (Re index 98–99), modern daylight LEDs perform best in the new equipment presented (Re 97–98). Existing LED lighting systems sometimes have very pronounced blue peaks and inequalities in the spectrum. The Re values are in the range between 45 and 82. Xenon light sources (such as flash units) show a balanced spectrum (Re index 87). Devices with fluorescent tubes (mercury vapour) achieve Re values of 65 to 84. The new devices ensure shadow-free and homogeneous illumination (deviation of the measured irradiance in the area < 5%) and the respective objectives (suitability for photography in the UV range, photography of entire boxes, low-reflection photography) are achieved. The paper shows some astonishing deficits in light quality in the photography and digitization of insects, which very probably also apply to other areas in biology and beyond. Digitization programs should always check the quality of their lighting and incorporate better light sources if necessary. LEDs with daylight quality are readily available and represent a high-quality alternative. Capturing colour patterns in the UV range enables the documentation of a biologically essential component of the electromagnetic spectrum. It should therefore play a far greater role in digitization programs as well as in ecological analyses.
Digital photography plays an outstanding role in many areas of biology in general and entomology in particular. For example, it is indispensable in the description of species and in morphology, but it also plays an essential role in the digitization of collections in museums (
In this article, some common and widely used light sources that play an important role in the photography and digitization of insects are spectrally measured. These, in particular, include mercury vapour fluorescent tubes, which are used in reproduction systems and light boxes. In recent years, LED lamps have become increasingly established in many areas, although they vary considerably in terms of energy efficiency and light quality. Commercially available flash units also continue to play an important role in macro photography, as they are considered to be true to colour (
Subsequently, three lighting devices that I have developed and tested in recent years are presented. These are (1) a light cylinder for photographing individual moths and other insects (upper- and underside) in the VIS (visible) and UV range, (2) a light box in which complete insect drawers can be photographed and (3) a light hemisphere, which was developed primarily for low-reflection photography of liquid surfaces (with insect samples). In addition, some recommendations on camera equipment and settings are given (Box
| The most suitable camera settings vary depending on the objects and also the purpose for which the images are to be used. However, there are many aspects that are similar for the vast majority of applications. When choosing a camera, it is important to consider whether the results are primarily to be produced quickly and inexpensively, or whether the focus is on high-quality results and long-term applications or archiving. For high-quality images, good quality SLR cameras, but above all mirrorless cameras and potentially also industrial cameras, should be considered. When it comes to lenses, the choice falls on macro lenses with very good optical properties, i.e. with low vignetting, low distortion and low chromatic aberration. The choice of focal length depends on the desired working distance. When photographing whole drawers, the distance to the sensor is already 124 cm with a focal length of f = 60, so that a longer focal length is not practical. In the field of studio photography and non-moving objects, shutter speed does not play a decisive role, and old manual lenses can also be used, as these can usually be connected well to mirrorless cameras using adapters and often have excellent optical properties. |
| Modern cameras offer a wide range of setting options, of which only the most important can be mentioned here. I always work in manual mode, where the aperture and shutter speed are set manually. The best resolution is achieved with an open aperture, the greatest depth of field with a closed aperture. In stacking photography, an almost open aperture is normally used. If a prepared moth or butterfly (which tends to be a “two-dimensional” object) is photographed instead of a truly three-dimensional object (such as a beetle), a single shot is sufficient for many purposes to achieve good results. In this case, a medium aperture should be selected so that a sufficient depth of field is guaranteed without compromising the absolute resolution too much. For the devices presented, an aperture of f/16 was selected in the photo cylinder, an aperture of f/5.6–8 when photographing insect boxes at a great distance and an aperture of f/11 when photographing ethanol samples. The correct exposure time results from the selected sensitivity (ISO setting) and aperture setting. ISO values with the lowest noise are recommended, i.e. low values of 100 or below. In order to fully standardize the exposure, it is necessary to use a standardized grey card or reflectance standards ( |
| A scale should be shown in each image. Overall, it is recommended to use a scale that is as uniform as possible – 10 mm is generally recommended for lepidopterans. The position of the scale in the image should also always be the same. Both of these factors facilitate subsequent automated evaluation by algorithms. The chessboard-like scale shown in Fig. |
| In many digitization programs, e.g. at the Natural History Museum in London (NHMUK), labels are recorded together with the insect. This has the advantage that there is no confusion and the insect and label are always clearly assigned. The disadvantage, however, is that the process tends to be time-consuming, as all the individual parts have to be arranged so that they lie in one focal plane. I therefore work with separate images of object and label in the case of non-standardized labelled animals. For insects that are recorded in modern and standardized campaigns, recording the label is obsolete because all location data is already recorded in a database and time and storage costs can be saved without loss. |
CIE International Commission on Illumination
CRI Colour Rendering Index
LED Light emitting diode
NHMUK Natural History Museum, London, UK
RGB Red, Green, Blue colour space
UV Ultraviolet
VIS visible (for humans)
For this paper, spectral measurements were carried out and documented using a Blue Spec Cube spectroradiometer from JETI (Jena, Germany). This was done in a similar way to that described by
Quantitative measurements were also carried out in all three devices to check the uniformity of the lighting. Uniformity of illumination is of obvious importance; in cinema projectors, for example, the ST 431-1:2006 standard stipulates that the peripheral areas must have at least 75% of the brightness of the centre. If images are to be evaluated quantitatively, all image areas should be evenly illuminated in order to prevent measurement artifacts. For the evaluation, one measurement was taken in the centre and four measurements in the respective corners. A total of nine measurements were carried out in the light hemisphere due to the increase in brightness observed at the edges. Suppl. material
Various full-frame mirrorless cameras from Sony are used for photography, but products of similar quality can also be used for the applications described. For photographing individual insects, I use a Sony Alpha 7 camera (24.3 megapixels), which was modified according to the specifications of
Moths and other insects are photographed from above (also with the other devices), with a camera that can be raised and lowered on a repro stand (Kaiser RTX tripod). The camera used is a modified Sony Alpha 7 camera (24.3 megapixels) (see above). The basic body of the light cylinder was commissioned from a locksmith. It consists of a cylindrical body made of sheet zinc with a door with a diameter of 45 cm and a height of 35 cm (Fig.
| When choosing the colour for the background of objects, there seems to be a lot of choice. For example, the contrast to the object can be maximized or the background can have a colour that does not occur in the object ( |
Light cylinder for VIS and UV photography: a. Reduced overview; b. Overview with open door (Scale: 10 cm); c. Structure of the lifting platform and three UV flash units in the cylinder. d. Interior view of the lighting on the ceiling; e. Moth with scale; f. Moth underside during photography of the proboscis length (or photography of labels) on polyethylene plate.
The light box for insect drawers (Fig.
As a cost-effective, lightweight and easy-to-make alternative to the light cylinder, the possibility of working with polystyrene hemispheres was tested. These are available in sizes with a diameter of up to 50 cm as decorative material (Rayher no. 3306400). Polystyrene has good reflective properties, including in the UV range. However, the material is translucent, so that on the one hand light escapes from the hemisphere and on the other hand light can enter from outside. Therefore, the hemisphere was covered with aluminum foil on the outside (Fig.
Light hemisphere. a. Overview with door open (lifted with pull rope); b. Overview with door closed (Scale 10 cm); c. Interior view with metal ring and LEDs; d. Interior view with metal ring hidden to avoid strong reflections on liquid; e. View of the photographed ceramic bowl with insects from ethanol sample; f. Hemispheres in different formats: diameter 30 cm and 50 cm.
Table
Quality of the light sources, based on spectral measurements in three museums (Berlin, Jena, London). The CIE Ra index (Colour Rendering Index) is based on the correspondence with 8 exemplary colours. The more meaningful and stricter CIE Re index is based on the correspondence with 16 exemplary colours.
| Source | Colour temperature ca. | CIE Ra | CIE Re | |
|---|---|---|---|---|
| Daylight | Sun/overcast/daytime | 5500–6500 K | >99 | >98 |
| Museum | ||||
| Ceiling lights in working areas | Mercury vapour | 3000–3600 K | 76–87 | 64–84 |
| Exhibition lights (Berlin, London) | LED | 2800–3100 K | 79–83 | 72–78 |
| New insect digitisation devices | ||||
| Light cylinder | Lumtronix Linear Z daylight LEDs | 6100 K | 99 | 98 |
| Light box | Lumtronix Linear Z daylight LEDs | 6400 K | 99 | 98 |
| Light hemisphere | Lumtronix Pro Sunlike | 4600 K | 98 | 97 |
| Insect digitisation devices | ||||
| Arc lamp (Xenon) | same as flash | 6200–6500 K | 91 | 87 |
| Mass digitisation (Berlin) | LED | 3900 K | 87 | 82 |
| Leica Z16 microscope (Berlin) | LED | 6900 K | 76 | 66 |
| Suprascan Quartz (London) | LED | 4100 K | 58 | 45 |
| Kaiser Repro Light (Berlin) | Mercury vapour | 4900 K | 88 | 81 |
| SatScan (London) | Mercury vapour | 3800 K | 76 | 65 |
| SatScan (Berlin) | Mercury vapour | 3600 K | 85 | 76 |
| Light Box with circle tube | Mercury vapour | 4700–5300 K | 79–87 | 70–84 |
Information on distances between camera and object as well as on the uniformity of the distribution of light in the three lighting devices. Max. dev = maximum observed deviation of measurements.
| Distance | Irradiation (W m-2) | Pixel in tif image (R=G=B) | ||||||
|---|---|---|---|---|---|---|---|---|
| filter wheel – insect | sensor – filter wheel | Centre | Range | Max. dev. | Centre | Range | Max. dev. | |
| Light cylinder (with diffusor) | ||||||||
| Scale 1:1 | 9.4 cm | + 18.5 cm | 16.58 | 16.39–16.55 | 1.1% | 210 | 210–211 | 1.0% |
| Scale 1:1.5 | 14.4 cm | + 18.5 cm | 16.22 | 16.22–16.34 | 0.8% | 219 | 219–222 | 1.4% |
| Scale 1:2 | 18.0 cm | + 18.5 cm | 16.61 | 16.61–17.23 | 3.7% | 225 | 225–231 | 2.7% |
| Scale 1:2.74 | 26.0 cm | + 18.5 cm | 18.19 | 18.19–18.68 | 2.7% | 227 | 227–232 | 2.2% |
| Scale 1:4.74 | 41.0 cm | + 15.4 cm | 21.05 | 20.76–21.25 | 2.3% | 239 | 235–239 | 2.7% |
| Light box (no diffusor) | ||||||||
| – | 124 cm | 10.97 | 11.12–11.48 | 4.7% | – | |||
| Light hemisphere | ||||||||
| ∅ 16 cm | – | 36 cm | 11.28 | 11.52–11.83 | 4.9% | – | ||
| ∅ 32 cm | – | see above | see above | 12.79–13.31 | 20.0% | |||
The light cylinder ensures shadow-free illumination in the VIS range (Fig.
Comparison of photographs taken in with different light sources. The images were standardized in terms of white balance and exposure (white reflectance standard set to RGB 241/241/241). They were saved in tif format but otherwise not modified. a. Shot in daylight (overcast sky); b. Shot in light box; c. Shot with SatScan (device in Berlin); d. Shot with Xenon flash. Note that monitors cannot reproduce all stored colour information of the Adobe RGB colour space and general limitations when reproducing images on displays and in print. See Suppl. material
The light box enables evenly bright and shadow-free illumination in the insect drawer (Fig.
The light hemisphere also enables shadow-free illumination (Fig.
Measuring the quality of light sources has, on average, revealed a significantly worse light quality than I expected. It could be argued that further problems can occur during subsequent colour management, e.g. due to the possible incorrect and distorted display of colours in monitors. Suitable lighting can certainly not prevent such errors, but is not a source of error itself. Apparently, too little attention has been paid to the quality of light sources in photography and digitization. For example,
The measured LEDs vary considerably in terms of light quality. Satisfactory values are achieved in the exhibition area of museums, where LEDs are used in which the blue component is significantly reduced for conservation reasons and which therefore appear ‘warm white’. Here, the focus is not on perfect colour reproduction – as is the case with street lighting, for example – but on protecting sensitive objects. Surprisingly poor Re values are achieved by the Supra Quartz Scanner at the NHMUK in London and the Leica Z16 microscope at the Museum of Natural History in Berlin. In both cases, these are LEDs with a strong blue peak and a pronounced deviation from the daylight spectrum. It is difficult to understand why LEDs of this quality, which are unsuitable for colour reproduction, are used in otherwise expensive devices. Flash units perform very well in terms of colour quality with CIE Re values of almost 90 and can be used without any problems. Extremely good CRI values are only achieved by the daylight LEDs presented here. Re values of >90 or >95 are now available on the market for a number of LEDs according to manufacturer information. In more recent works such as by
All three lighting devices serve their respective purpose, although each has a different basic shape (cylinder, cube, hemisphere). The basic principle is to illuminate from many sides, which is made possible by scattering the light from the inside. The aim of the lighting is to obtain as high a proportion of scattered light as possible, with perfect conditions occurring outdoors on a cloudy day. This ensures that there are no unwanted shadows that can lead to problems in image evaluation. The devices presented do not, however, enable reflection-free photography, but in the light cylinder and in the light hemisphere these “specular” reflections are greatly reduced by the use of diffusers (Fig.
In all devices, light is not allowed to leak out in large quantities (energy efficiency and uniformity) or from the outside to the inside, for example when the devices are operated in a room with windows. Another approach is taken by
Photographs taken in the light cylinder: a. Male and b. female of Gonepteryx rhamni (VIS and UV); c. Male and d) female of Lycaena alciphron gordius (each in the VIS and UV range). In the UV range, patterns become visible in the males of both species through UV photography that cannot be perceived by humans.
Photographing entire drawers is useful in order to begin recording collections where they do not yet exist. This appears to be an important option, especially in smaller museums, because the resources for individual digitization are not available. Here, it is often the case that hardly any changes occur in historical collections over many years, so that the images represent the current status for a long time. Another advantage is that negative changes are documented, for example due to pest infestation or theft. For parts of the collection, however, where a lot of work and sorting takes place, photographing entire boxes is less useful. Individual recording is particularly suitable for such parts of the collection. The light hemispheres are a simple, inexpensive alternative that anyone can build. Their appearance, however, has a more “DIY” character, but this does not have to be a problem. The hemispheres, like the light cylinder, are relatively cramped. For some objects, the height of the hemisphere itself is not sufficient, so the sphere can be placed on a base, which can be made of polystyrene or wood wrapped in aluminum foil, for example. Shielding the direct LED radiation using aluminum adhesive tape enables low-reflection photography of wet samples. Further work will have to show whether, and to what extent, it is possible to identify material based on the photographs. The high resolution achieved in any case allows the taxa to be identified by order, but usually at a much higher taxonomic level.
UV photography enables the documentation of colour patterns in insects and other objects that go well beyond the usual RGB colour space. This is sometimes impressive, as in the case of male brimstone butterflies (Pieridae: Coliadinae) and many copper butterfly (Lycaenidae: Lycaeninae) species, where patterns previously not visible to humans appear (Fig.
a. UV photography provides an additional fourth colour channel (UV), in addition to the usual channels B = Blue, G = Green, R = Red; b. Normal RGB image; c. False colour image with colour information GBU, shown in the usual RGB colour space; d. Measured relative sensitivity of a camera sensor in four colour channels (modified Sony Alpha 7) compared to the relative absorption of the cones in a bird’s eye (Eurasian Blue tit:
One reason that the recording of brightness information in the UV range has been neglected is certainly that the same systems that are used for photographing ‘normal’ cultural assets simply cannot be used. The recording of UV data is currently (still) a ‘tinkering task’, and efficient concepts and tools for the evaluation of data seem to be largely lacking. The necessary cameras that are comparable in comfort and price to normal RGB cameras cannot yet be bought off the shelf. However, as the example above also shows, it is not at all impossible to produce high-quality UV images with reasonable financial means (
The present study shows some astonishing deficits in the quality of light commonly used for the photography and digitization of insects, which very likely applies to other areas in biology and beyond. Future studies should show whether and what effects poor light quality has on the results of e.g. lightness measurements of insects in quantitative studies. Based on the insight of this study, digitization programs should fundamentally check the quality of their lighting and then install better light sources if necessary. LEDs with daylight quality are now readily available and they represent a long-lasting, ecologically acceptable and high-quality alternative to existing systems with poor light quality. The additional costs compared to ‘normal’ LEDs are insignificant compared to the higher image quality achieved. Spectral measurements can provide final certainty, because the quality of light cannot be assessed by eye. The recording of colour patterns in the UV range enables the documentation of a biologically essential component of the electromagnetic spectrum. The UV range should therefore play a much greater role in digitization programs, as well as in ecological analyses, than it has done until now.
I would like to express my great thanks to Daniel Veit and the Max Planck Institute for Chemical Ecology Jena for continuous and great help in developing and building the systems, as well as for advice on questions regarding the choice of suitable LEDs and programming the control for the filter wheel. Colleagues in London and Berlin enabled me to take measurements – Berlin: Bernd Schurian; London: Geoff Martin, Alberto Zilli and David Lees. JETI (Steffen Göhrlich and Arnd Hinze) in Jena allowed me to carry out measurements of the camera on their monochromator. I am also very grateful to Eric J. Warrant (Lund), who checked and improved the content and language of the entire manuscript and reviewed it – many thanks also to the second reviewer for his valuable comments. Robert Brunner (Jena) kindly checked optical-technical aspects in the manuscript. I would like to thank Egbert Friedrich (Jena) for important references and thematic discussions. Yenny Correa performed some measurements with the “LEPY” algorithm. She, Dennis Böttger (Jena), Klaus-Rudolf Lunau (Düsseldorf) and two reviewers provided valuable comments on an earlier draft of the manuscript.
Light cylinder: Measurements at different scales and light homogeneity; Light box & Light hemisphere: Light homogeneity
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Spectral measurements and Colour Rendering Index (CRI)
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Spectral measurements and Colour Rendering Index (CRI)
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Moth box and colour chart images in different lighting settings
Data type: pdf