Riddle, Jeanne M.

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Henry Ford Hospital Medical Journal Volume 33 Number 1 Article 2 3-1985 Electron Microscopic Examination of Various Types of Ancient Specimens Jeanne M. Riddle Follow this and additional works at: https://scholarlycommons.henryford.com/hfhmedjournal  Part of the Life Sciences Commons, Medical Specialties Commons, and the Public Health Commons Recommended Citation Riddle, Jeanne M. (1985) "Electron Microscopic Examination of Various Types of Ancient Specimens," Henry Ford Hospital Medical Journal : Vol. 33 : No. 1 , 3-8. Available at: https://scholarlycommons.henryford.com/hfhmedjournal/vol33/iss1/2 This Article is brought to you for free and open access by Henry Ford Health System Scholarly Commons. It has been accepted for inclusion in Henry Ford Hospital Medical Journal by an authorized editor of Henry Ford Health System Scholarly Commons. Henry Ford Hosp Med J Vol 33, No 1, 1985 Feature Articles 41 43 Electron Microscopic Examination of Various Types of Ancient Specimens Jeanne M. Riddle, PhD* 48 57 fei 62 64 Paleopathology (the study of disease in ancient re-mains) is an important and interesting area of inves-tigation. Through these studies, we can: 1) learn the time frame in which specific diseases emerge within identified cultures, 2) determine if the tissue response to the pathologic alteration remains constant over long periods of t ime, and 3) assess whether or not modern populations share the same diseases with their ancient counterparts. On the morphologic level, tissue samples obtained from ancient humans were first examined histologically with the light microscope. More recent literature con-tains a l imi ted number of ultrastructural and top-ographic studies in which samples removed from or associated with mummif ied ancient bodies were inves-tigated with either the transmission electron micro-scope or the scanning electron microscope (1-4). Because it is necessary to fix tissues and cells immedi-ately in order to preserve the full range of their ultra-structural features, f ine structural details of ancient specimens are f requent ly compromised at the in-creased resolution and magnification levels possible with the transmission electron microscope. However, sufficient morphologic characteristics remain in mum-mified tissue specimens to identify positively classes of blood cells, specific types of fibers such as collagen, and characteristic developmental stages of parasites. The addition of the scanning electron microscope to the armamentarium of the paleopathologist has increased the types of ancient samples that can be studied. Using this instrument, we can gain for the first t ime a view of the topography of these materials in apparent three-dimension. Over the past 12 years, I have had the opportuni ty to study a variety of samples col lected f rom Egyptian jnummies autopsied under the auspices of the Pa-leopathology Association, an international organization comprised of basic scient ists, physicians, anthro-pologists and archeologists, founded by the late T. ^idan Cockburn, M D , Detroit, Michigan. Additional •"sect remains were obtained from Peruvian mummy bundles during their dissection by James M. Vreeland, IJ"' archeologist, LJniversity of Texas, Aust in, Texas, elected examples f rom these samples wil l be pre-sented in this publ icat ion. Background Information on the Various Mummies Egyptian mummies, obtained f rom the Pennsylvania University Museum, are identif ied by an acronym PUM and a Roman numeral which indicates the order in which the mummies were received and autopsied (I I , III and IV). PUM II was carbon dated at 170 BC ± 70 years. This mummy was an adult man, 35 to 40 years old with a height of 5 feet 4 inches. He was thought to be an important person, possibly a priest. The method used for his embalming was classic, that is, his organs were preserved, the body was wrapped in l inen, and the mummy was placed into an elaborately decorated sar-cophagus (5). PUM III was carbon dated at 835 BC ± 70 years. This mummy, an adult woman approximately 35 years o ld , was 5 feet 1 inch tall wi th a calculated weight of 95 pounds. In contrast to PUM I I , this mummy was evis-cerated per ano, wrapped in pieces of medium quality l inen, and was not contained with in a sarcophagus. Tissue preservation of this body was generally of poor quality (6). PUM IV was dated at the first or second century AD (Roman period). This mummy was a male chi ld, 8 to 10 years in age and 3 feet 5 inches tall. This mummy had also been eviscerated per ano. Its wrappings consisted of linen strips, and the body was dressed in a numbero f full or shorter length shroudlike garments. The body tissues of this mummy were also severely degenerated (6). Insect specimens were also collected from three Pe-ruvian mummy bundles, the major form of burial for central and south coastal Peruvian cultures. Each bun-dle contained a mummy in a flexed body posit ion. The mummies were wrapped in successive layers of plain-weave and/or patterned textiles. Layers of the textiles were often alternated with other materials such as seed Submitted for publication: April 19, 1985 Accepted for publication: April 23, 1985 'Research Electron Microscopy Laboratory, Henry Ford Hospital Address reprint requests to Dr Riddle, Research Electron Microscopy Labo-ratory, Henry Ford Hospital, 2799 W Grand Blvd, Detroit, Ml 48202. Riddle cotton, leaves, grass or hides. Each of these mummy bundles was associated wi th three distinct cultural complexes: Paracas (86 AD), Epigonal (1231 AD), and Huancho (1240 AD). The bodies contained with in the bundles were those of a 40- to 60-year-old woman, an 18- to 20-year-old woman, and a 12-year-old chi ld, re-spectively (4). Ancient Specimens Surveyed Outer wrappings Wrappings collected from the Egyptian mummy PUM II consisted of a fabric constructed f rom relatively fine threads (Fig 1). When individual threads of this cloth were surveyed with the scanning electron microscope, we found that they were composed of slender, straight fibers that showed joint l ike markings (Fig 2). These fibers showed a topographic morphology that was identical wi th reference flax fibers. By studying this ancient textile with the scanning electron microscope, we were able both to view its pattern of weaving and to confirm that it was l inen, the traditional cloth in which Egyptian mummies were wrapped. Fig 1 Scanning electron micrograph of the linen wrappings removed from PUM 11 X 38, Fig 2 Individual flax fibers which make up the threads of the linen wrappings of PUM II X 800. During the unwrapping process on PUM I I , we found a small ball that was composed of whit ish fibers incrusted with possibly dried unguents. On gross observation, this appeared to be a ball of cotton wrapped between two small pieces of l inen. Examination of the individual whitish fibers by scanning electron microscope showed that they were straight fibers that had a repeating/ inherent twist (Fig 3), a morphology that corresponded exactly to the three dimensional appearance of refer-ence cotton fibers. Electron microscope confirmation that this ball of material was indeed cotton was an important piece of informat ion, since cotton had not previously been found in the western wor ld before the t ime of Christ. The earliest identif ication of cotton until its discovery among the linen wrapping of PUM II was from a Roman grave of about 200 AD. Human blood cells During the autopsy of PUM I I I , a small plaquelike mass of brown, granular material that had adhered to the inner aspect of the occipital bone was discovered anO removed for further study. This material was originally Electron Microscopy of Ancient Specimens Fig 3 n wrappings of individual fibers of cotton showing their inherent twist removed from the cotton ball found on PUM II x 650. A'e found a s incrusted 3servation, d between ! individual pe showed repeating, responded ;e of refer-)nfi rmation on was an j n had not before the :otton until ' U M II was je l ike mass ;red to the overed and, IS originally thought to be brain tissue because of its anatomic loca-tion, but after it was rehydrated, sectioned and exam-ined by light microscopy was found to contain numer-ous peripheral blood elements. While it is quite common to observe well-preserved erythrocytes in the tissues of mummif ied bodies, leu-kocytes are rarely found . This mass of material, which was either an antemortem hematoma or a postmortem blood clot, gave us the opportuni ty to observe both the topographic features and the quality of ultrastructural preservation of human blood cells over 2,500 years o ld . Some of the ancient erythrocytes retained their bicon-cave shape (Fig4) and still contained an electron-dense material, presumably hemoglobin (Fig 5). The whole erythrocytes were first viewed with the scanning elec-tron microscope; then the same cells were studied in ultra-thin sections w i t h the transmission e lect ron f'licroscope. It was possible to identify various classes of leukocytes, ie, lymphocytes, neutrophils, and eosino-phils. The l imit ing plasma membrane of these cells was generally intact. Their nuclei were distinct and showed characteristic distributions of heterochromatin and eu-chromatin. The cytoplasm of leukocytes recognized as lymphocytes showed swollen mitochrondr ia and r i -bosomes (Fig 6). The cytoplasm of neutrophils showed numerous typical membrane-bound neutrophi l gran-ules (Fig 7). Hair Although the soft tissues of PUM IV were degenerated, the hair on the head of this mummy remained intact. When we examined some of these hairs by scanning e l ec t r on m ic roscopy , we f o u n d that the m i c r o -architecture was preserved. The prominent scalelike plates (epicuticles) were seen to have retained their transverse orientation and regular spacing (Fig 8). Insects Adult insects and their larval forms are sometimes f o u n d be tween the w rapp ings of m u m m i e s and mummy bundles or even associated with the fleshy parts of the bodies. Fig 4 Scanning electron micrograph of an ancient erythrocyte—note central concavity x 14,500. Riddle Sectioned ancient erythrocytes from PUM III show internal electron-dense hemoglobin. Black material on surface is layer of deposited gold-palladium added to perform SEM studies x 15,000. Ancient lymphocytes showed a distinct nucleus, swollen mitochondria and ribosomes within their cytoplasm x 23,000. I \^T|Pg| Ancient neutrophils contained a distinct nucleus and well-pre-served neutrophilic granules within their cytoplasm x 15,000. Using the eas on the mouthpar served in in most in tive ident insects. Tl IV were id known as (Linnaeus A variety c Peruvian i distinct s beetles (Fi tron micrc featu re of was displa proach (Fi The applic samples c associated Examinatic Fig 8 Topographic details of a single hair removed from the head of PUM IV X 630. '^ so.ca|led"dr Electron Microscopy of Ancient Specimens Using the scanning electron microscope, different ar-eas on these immature and mature insects such as their mouthparts, appendages, and integument can be ob-served in detail (Fig 9). Compilation of this information in most instances provides the necessary clues for posi-tive identification and classification of these ancient insects. Those associated with the fleshy parts of PU.M IV were identified as members of the Anobiidae family, known as the drugstore beetle, Stegobium paniceum (Linnaeus). A variety of insect forms were found associated wth the Peruvian mummy bundles that we investigated. Two distinct species of flies and five dist inct genera of beetles (Fig 10) were identif ied by using scanning elec-tron micrographs. In addi t ion, a previously unknown feature of the integument of the Call iphorid puparium was displayed using this three-dimensional survey ap-proach (Fig 11). Discussion The application of electron microscopy to the study of samples collected f rom various mummies and their associated wrappings has produced new information. Examination of blood cells found in an Egyptian mummy e head of , Fig 9 A so-called "drugstore beetle" removed from the soft tissues of PUM IV x 125. Fig 10 Adult beetle, Anobiidae Stichtoptychuson near, found within the Paracas mummy bundle x 40 (Photo credit, Paleopathology Newsletter 1982;39:5-9). and studied by a con t inuum of microscopy (l ight, t ransmiss ion and scanning e lec t ron microscopy) showed that erythrocytes and leukocytes over 2000 years old had the same ultrastructural features as their modern counterparts. Even though the time span of 28 centuries represents a brief period on the total evolu-tionary scale, our study showing the overall constancy of the construction of peripheral blood elements rep-resents an important and previously unknown f inding. The ability to analyze fabrics using the scanning elec-tron microscope also provides a more detailed way of confirming the method of their construction (pattern of weaving) as well as the basic fiber used than does the more conventional examination wi th the light, dis-secting and polarizing microscope. Finally, identifying insects found either in the flesh or contained between the various wrappings gives us in-sight into the effectiveness of different preservation processes (embalming techniques) and burial practices in general. We should continue to use modern technology and newly developed instruments to study ancient speci-mens in order to further our understanding of earlier Riddle Henry Fon Vol 33, No cultures. However, we should be aware that these samples, which offer a unique opportuni ty to explore our past, must be used judiciously as a nonrenewable resource. Fig 11 A previously undiscovered feature of the integument of a puparium of the fly Calliphoridae Cochliomyia macellaria (Fabricus) recovered from the Paracas mummy bundle x 132 (Photo credit, Paleopathology Newsletter 1982;39:5-9). References Riddle JM. A survey of ancient specimens by electron micros-copy. In : Cockburn A, Cockburn E, eds. Mummies, disease and ancient cul tures. Cambr idge, England: Cambridge University Press, 1980:274-86. Shipman P. Appl icat ions of scanning electron microscopy tO| taphonomic problems. In : Cantwell A M , Gri f f in JB, Rothchild NA, eds. The research potent ial of anthropological museum! col lect ions. Ann NY Acad Sci 1981;376:357-85. Horne PD. Optical and electron microscopy. In : Hart C D , edJ Disease in anc ien t m a n . T o r o n t o , Canada: Clarke I rw in 1983:191-8. Riddle JM, Vreeland JM Jr. Identi f icat ion of insects associated w i t h Peruvian m u m m y bundles by using scanning electron microscopy. Paleopathology Newsletter 1982;39:5-9. Cockbu rn A, Peck W H , Barraco RA, Reyman TA. A classic! mummy: PUM I I . In : Cockburn A, Cockburn E, eds. Mummies, disease and ancient cultures. Cambridge, England: Cambridge University Press, 1980:52-70. Reyman TA, Peck W H . Egyptian mummif icat ion wi th evisceration I per ano. In : Cockburn A, Cockburn E, eds. Mummies, disease] and ancient cultures. Cambridge, England: Cambridge Univer-sity Press, 1980:85-100. Dr Fra Richmoni The flowc took place emerging I concern,w medical pr century wh and hospii endowed i clinical sk and house served dur demic mec Halsted, ar ings left ir fessionals. spirit and Sir William influenced Chairman ( Hospital, V, For insight nection wi principal si personal ( memory ar My apprec several bo death. Th€ Cushing's and Other Sladen left tations, ch are of greal ferret out ophies wh possibly in ""rankj. Sia River. His f 3n Aide-de O o. HOM S'gnments Frank Slad( some time preference probably n University, graduated played > 

Electron Microscopic Examination of Various Types of Ancient Specimens

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Electron Microscopic Examination of Various Types of Ancient Specimens

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