MATERIAL

The UV filters: Four Schottt monochromaticUV interference filters transmitting at 313, 333,365, and 385 nm, respectively; and for comparison,two broad band UV filters (Schott UG 1 and BG12) were used (Fig. 2). The filters were placedin front of the quartz lens in a light-tight mount.

The quartz lens: The quartz lens (5,4/135Astro, West-Berlin, Germany), transmitting downto 200 nm, was fitted to the bellows of a Hassel-blad 1000F camera (Fig. 3a and b).

The UV image converter: The invisible UV-image was visualized with the RCA 7404 UV-image converter tube (Fig. 3a).

The double-beam focusing device: The UVimage converter is not indispensable for focusing,

Presented at the Twenty-seventh Annual Meet-ing of The Society for Investigative Dermatology,Inc., Chicago, Ill., June 28, 1966.

* From the Department of Dermatology, ** Uni-versity Central Hospital, Snellmaninkatu 14, Hel-sinki 17, Finland and from the Department ofPhotography, 5*5 University of Helsinki, Helsinki,Finland.

t Jenaer Glaswerk Schott & Cen., Mainz, Ger-many.

351

Vol. 47, No. 4Printed in U.S. A.

when there is no need for successive operation atseveral UV wavelengths (cf Fig. 1). When thefocal plane for a certain monochromatic filterhas first been empirically determined, focusingcan he exactly repeated with a simple double-beam focusing device (Fig. 3b) operating on theprinciple of triangulation, i.e. the focal plane isreached when the two focusing beams converge(Fig. 3c). The double-beam focusing is also suitedfor close-ui) photography in visible light.

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Fme. 1. Schematic representation of the refrac-tion through a lens. The shorter the wavelength,the greater is the angle of refraction, and thebroader the band of radiation, the greater is thedispersion and the blurring of the image in theplane of the film. Note that UV lies outside therange of correction of ordinary camera lenses.

Fie. 2. TransInission characteristics of a mono-chromatic UV filter (a) and of a conventionalbroad-band UV filter (b). The significnnt band-width of the monochromatic filter is less thanlSnm.

THE JoeaNAI. OF INVESTIGATIVE DEEMATOLOGY

Copyright 1966 by The Williams & Wilkins Co.

MONOCHROMATIC ULTRAVIOLET-PHOTOGRAPHYIN DERMATOLOGYTM

KIMMO K. MUSTAKALLIO, M.D.55 AND PAAVO KORHONEN55TM

Photography with reflected ultrnviolet rayshas been proposed for accentuated differentia-tion of pigmentation and increased definition (1).The usefulness of this technic has been limitedby the difficulty in focusing the invisible DV-image and by the disadvantages inherent in or-dinary glass lenses and conventional broad-bandDV filters. Glass lenses cannot be used below350 nm (mp) and they absorb more than two-thirds of the radiation already at 365 nm. Fur-ther loss in intensity is caused by reflection, andthe more glass surfaces in a lens, the less is thetransmission for DV (2). Furthermore, the lensesof ordinary cameras are not corrected for DV.The DV rays of widely differing wavelengthtransmitted by conventional broad-band DV fil-ters are refracted along different paths to vary-ing depth (Fig. 1). This dispersion causes blur-ring of the image.

These obstacles in UV-photography of theskin have been eliminated by using an equip-ment consisting of monochromatic DV filters, aquartz-lens camera, an DV image converter,and an electronic Xenon flash unit.

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352 THE JOURNAL OF INVESTIGATIVE DERMATOLOGY

Fm. 3. The equipment used fer monochromatic DV photography: (a) General viewshowing the DV image converter mounted on the camera and the electronic Xenon-flashused for close-up photography; (b) A detail showing the quartz lens, a monochromatic DVfilter (not in proper place to show the lens),, and the two lamps for the double-beamfocusing; (c) When the two light spots on the skin converge, the right focal plane is reached.

The UT-sources: For searching and focusing aPhilips MLTJ 300 W DV lamp was used. Forphotographic exposure an electronic Xenon flashequipment having a power output of 400 to 1000IV per second was fully sufficient for the filtersused.

The film: Ilford Gelochrome (160 ASA) withordinary developers and routine laboratory proc-essing gave good results.

ILLUSTRATION OF APPLICATIONS

Melanin pigmentation and it s disorders. Themonochromatic DV technic with a 365 nm filter

is ideal for accentuated differentiation of mela-nin pigmentation, whether it is increased (Fig.4) or diminished (Fig. 5).

Resolution of fine details of the shin surface.When monochromatic DV rays are used inclose-up photography, the definition of the sur-face texture of the skin approaches the levelreached by the best imprint or replica technics(Fig. 6). For comparison the same area of skinwras photographed with the quartz-lens camerausing a green filter, a broad-band TJV filter, andthree different monochromatic DV filters (Fig

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MONOCHROMATIC UV-PHOTOGRAPHY 353

FIG. 5. Vitiligo in a very blonde woman is hardly discernible in the conventional photo-graphs of the face, neck, and hands, but becomes strongly accentuated at 365 nm. Note alsothe better definition of the details of the skin surface in the monochromatic UV pictures.

a bFIO. 4. Dark-season appearance of the face of

a blonde woman (a) in polychromatic hght (theright half) and (b) at 365 nm. The freckles, al-most invisible in (a), stand out in (b) as stronglyabsorbing black spots against the more reflectingnon-pigmented areas.

7). With the monochromatic filters the defini-tion of the fine surface details is superior, in-creasing and giving a three-dimensional appear-ance by decreasing wavelength.

DIscUSsION

the monochromatic rays penetrate to the samedepth and arc more regularly refracted and re-flected (Fig. 1). All these facts explain why themonochromatic IJY-rays produce an image ofbetter definition than broad-band lilY radiationor visible light and why it is better the shorterthe wavelength. On the other band, lilY radia-tion which does not penetrate deep enougb,being largely absorbed in the epidermis, doesnot visualize dermal events, such as sugilla-tions (Fig. 8) or erythema (Fig. 9).

Absorption: Without regard to the melanincontent, human epidermis absorbs below 250nm about 80 per cent of the radiation (4). Theskin looked almost black and differences in mel-anin pigmentation were hardly discernible, whena monochromatic Schott lilY reflectance filtertransmitting at 250 nm was used. lit will be in-teresting to relate the image at 270 nm to theurocanic acid content of the horny layer (5).

Three properties of radiation, penetration,absorption, and reflection, are to be consideredin relation to the monochromatic technic of BYphotography.

Penctrotion: A full-tluckness sheet of epi-dermis, separated by suction (3) from the volarforearm of white blonde persons, appears to fil-ter out below 290 nm all BY radiation emittedby the Beckman DB spectrophotometer. An in-crease of wavelength is associated by greaterpenetration. But the deeper the radiation pene-trates the greater is its dispersion and the dif-fusion of the reflected rays. This blurs down thedefinition of the photographic image. Further-more, in contrast to "polychromatic" radiation,

Fio. 6. Close-up view of a hand-flate at 365 nm.Original magnification 2:1. Note the three-di-mensional appearance and the sharply definedsweat pores.

354 THE JOURNAL OF INVESTIGATIVE DERMATOLOGY

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green BC 12 385 urn 365 urn 333 urnho. 7. Comparison of five different filters in close-up photography. The same area of

the dorsum of hand (note the hair in the middle) photographed through a green filter, abroad-band UV filter (BC 12), and three monochromatic UY filters at 385 um, 365 nm, and333 nm. The definition and the three-dimensional appearance increases from left to right.The pietures become darker because at shorter wavelengths more of the radiation is ab—sorbed and less reflected.

he. 8. Sugillatioos, clearly seen in the upperconventional photograph, cannot be detected at365 nm in spite of the very thin atrophic epidermis(lower picture).

355

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356 THE JOURNAL OF INVESTTGATIVE DERMATOLOGY

also the reflection, of monochromatic DV raysfrom the skin is more homogeneous than that ofbroad-band DV radiation or visible light. More-over, at shorter wavelengths a considerable pro-portion of the reflected radiatiort is polarized(7). The polarization also contributes to the su-

perior contrast and resolution of line details ofskin surface, which attains a three-dimensionalappearance in the monochromatic DV photo-graphs.

The monochromatic technic of DV photogra-phy lends itself also for the study of the re-flectance, and of absorption properties of light-screening agents at the DV range of solarspectrum. The photography of fluorescence com-plements it in such studies. For instance,acrylonitriles absorb at 365 nm more stronglythan red veterinary pctrolatum, but the latteremits a rather intense whitish fluorescence, i.e.transforms a part of the radiation to a light oflonger wavelength.

SUMMARY

Fio. 9. The rcticular erythema of systemic lupuserythematosus can he visualized by the conven-tional technic withoot any filters (upper) but notwith monochromatic DV radiation at 365 nm(lower). On the other hand, the hyperpigmentationaround the inflamed area can be detected only inthe lower pictore, showing also better the textureof the skin surface.

At longer DV wavelengths more radiation isreflected towards the camera to cause an imagewhich shows better contrast with areas wherethe radiation has been absorbed by melanin.The best results in the study of melanin pig-mentation have been obtained with the 365 nmfilter. A 410 or 430 nm filter should apply toearly detection and follow-up of jaundice, be-cause the degradation products of hemoglobinabsorb at this spectral area. Monochromatic DVphotography may also be suited for the studyof the epidermal distribution of drugs absorb-ing long DV radiation, such as chloroquiae,phenothiazines, and tctracyclincs (6).

Reflection: Not only the penetration, but

Monochromatic ultraviolet photography per-mits a new visualization of the epidermis. Itaccentuates differences in melanin pigmenta-tion, and it is ideal for close-up photography ofthe minute details of skin surface. The equip-ment required consists of monochromatic DVinterference- or reflectance-filters, a quartz-lenscamera, a broad-spectrum DV-lamp, an DV-image converter (for visualization and focusing),and an electronic Xenon flash unit.

REFERENCES1. Lunnon, R. J.: Direct ultra-violet photography

of the skin. Med. Biol. Illus., 9: 150, 1959.2. Dc Bruin, J. P.: Principles of ultraviolet light

and some of its apphcations ui photography.J. Biol. Photogr. Ass., 29: 53, 1961.

3. Kimstala, If. and Mustakallio, K. K.: In-vivoseparation of epidermis by production ofsuction blisters. Laneet, I: 1444, 1964.

4. Edwards, E. A., Finklestein, N. A. and QuimbyDuntley, S.: Spectrophotometry of livinghuman skin in the ultraviolet range. J. Invest.Derm., 16: 311, 1951.

5. HoráSek, J.: Die Liehtbarrierc der Epidermis.Derm. Wschr., 151: 887, 1965.

6. Mustakallio, K. K.: Tetraeyelines and deposi-sition of calcium. Laneet, II: 721, 1962.

7. Nicolam, L.: Deber die Wellenlkagenabhiingig-keit der Lichtpolarisation der menschliehen[Taut. PfiOeger Arch. Ges. Physiol., 270: 199,1960.

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24. Wynn, C. H. and Iqbal, M.: Isolation of ratskin lysosomes and a comparison with liverand spleen lysosomes. Biochem. J., 98: lOP,1966.

25. Olson, R. L. and Nordquist, R. E.: Ultramicro-scopic localization of acid phosphatase inhuman epidermis. J. Invest. Derm., 46: 431,1966.

26. Rowden, C.: Ultrastructural studies of kera-tinized epithelia of the mouse. I. Combinedelectron microscope and cytochemical studyof lysosomes in mouse epidermis and eso-phageal epithelium. J. Invest. Derm., 49: 181,1967.

27. Prose, P. H., Sedlis, E. and Bigelow, M.: Thedemonstration of lysosomes in the diseasedskin of infants with infantile eczema. J. In-vest. Derm., 45: 448, 1965.

28. Hall, J. H., Smith, J. G., Jr. and Burnett, S.C.: The lysosome in contact dermatitis: Ahistochemical study. J. Invest. Derm., 49:590, 1967.

29. Pearse, A. C. E.: p. 882, Histochemistry Theo-retical and Applied, 2nd ed., Churchill, Lon-don, 1960.

30. Pearse, A. C. E.: p. 910, Histacheini.stry Thea-retscal and Applied, 2nd ed., Churchill, Lon-don, 1960.

31. Daniels, F., Jr., Brophy, D. and Lobitz, W. C.:Histochemical responses of human skin fol-lowing ultraviolet irradiation. J. Invest.Derm.,37: 351, 1961.

32. Bitensky, L.: The demonstration of lysosomesby the controlled temperature freezing sec-tion method. Quart. J. Micr. Sci., 103: 205,1952.

33. Diengdoh, J. V.: The demonstration of lyso-somes in mouse skin. Quart. J. Micr. Sci.,105: 73, 1964.

34. Jarret, A., Spearman, R. I. C. and Hardy, J. A.:Histochemistry of keratinization. Brit. J.Derm., 71: 277, 1959.

35. De Duve, C. and Wattiaux, R.: Functions oflysosomes. Ann. Rev. Physiol., 28: 435, 1966.

36. Waravdekar, V. S., Saclaw, L. D., Jones, W. A.and Kuhns, J. C.: Skin changes induced by

UV irradiated linolenic acid extract. Arch.Path., 80: 91, 1965.

37. Nicolaides, N.: Lipids, membranes, and thehuman epidermis, p. 511, The EpidermisEds., Montagna, W. and Lobitz, W. C. Aca-demic Press, New York.

38. Wills, E. D. and Wilkinson, A. E.: Release ofenzymes from lysosomes by irradiation andthe relation of lipid peroxide formation toenzyme release. Biochem. J., 99: 657, 1966.

39. Lane, N. I. and Novikoff, A. B.: Effects ofarginine deprivation, ultraviolet radiationand X-radiation on cultured KB cells. J.Cell Biol., 27: 603, 1965.

40. Fukuyama, K., Epstein, W. L. and Epstein,J. H.: Effect of ultraviolet light on RNAand protein synthesis in differentiated epi-dermal cells. Nature, 216: 1031, 1967.

41. Daniels, F., Jr. and Johnson, B. E.: In prepa-ration.

42. Ito, M.: Histochemical investigations of Unna'soxygen and reduction areas by means ofultraviolet irradiation, Studies on Melanin,Tohoku, J. Exp. Med., 65: SupplementV, 10, 1957.

43. Bitcnsky, L.: Lysosomes in normal and patho-logical cells, pp. 362—375, Lysasames Eds.,de Reuck, A. V. S. and Cameron, M. Church-ill, London, 1953.

44. Janoff, A. and Zweifach, B. W.: Production ofinflammatory changes in the microcircula-tion by cationic proteins extracted from lyso-somes. J. Exp. Med., 120: 747, 1964.

45. Herion, J. C., Spitznagel, J. K., Walker, R. I.and Zeya, H. I.: Pyrogenicity of granulo-cyte lysosomes. Amer. J. Physiol., 211: 693,1966.

46. Baden, H. P. and Pearlman, C.: The effect ofultraviolet light on protein and nucleic acidsynthesis in the epidermis. J. Invest. Derm.,43: 71, 1964.

47. Bullough, W. S. and Laurence, E. B.: Mitoticcontrol by internal secretion: the role ofthe chalone-adrenalin complex. Exp. Cell.Res., 33: 176, 1964.