Clues to the Past: Further Development of the Comparative Plant Fiber

May 5, 1996 - The Comparative Plant Fiber Collection (CPFC), established to provide comparative plant fiber materials for the identification and ...
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Chapter 16

Clues to the Past: Further Development of the Comparative Plant Fiber Collection

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Kathryn A. Jakes Department of Textiles and Clothing, Ohio State University, 1787 Neil Avenue, Columbus, OH 43210

The Comparative Plant Fiber Collection (CPFC), established to provide comparative plant fiber materials for the identification and characterization of the fibers employed in prehistoric native American textiles, continues to be expanded. This report includes new information concerning morphological distinctions observed in plant fibers observed under scanning electron microscopy, the effect of treatment of the fibers in fiber processing, and the inorganic crystalline inclusions which the fiber products contain. These data may provide further distinctions among fibers within the four categories of plant fibers proposed as the results of previous work. The identification and characterization offibersemployed in textiles helps us reconstruct past methods of textile production and learn how ancient textiles were used. These data can contribute to studies of craft specialization and social differentiation.

Employing plant materials available locally or, perhaps, importedfroma distance, prehistoric native Americans produced a wide variety of textile products. Visual examination alone is sufficient to show that these textile products required sophisticated fabrication techniques. While coarsely made bags and mats have been recovered, fine yarns twined in intricate patterns are also seen in textiles from Hopewell (200 BC-500 AD) and Mississippian (1000-1500 AD) sites (1-10). Textiles are material objects; through their

0097-6156/96/0625-0202$12.25/0 © 1996 American Chemical Society

In Archaeological Chemistry; Orna, M.; ACS Symposium Series; American Chemical Society: Washington, DC, 1996.

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study, the materials and methods employed in their manufacture and the patterns of their use may be discerned. Textiles also serve as cultural objects, providing examples of dress and adornment, which can be used to reveal such factors as roles, status, and social differentiation. While the fabrication structure of some textiles recovered from prehistoric native American sites has been studied (5), extensive analysis of the fibrous components of these textiles has been hampered by the lack of a comparative collection of fibers representative of those employed in textile production. Identification and characterization of fibers in textiles has been limited, in some cases, to the description of the fibers as "bast" or "vegetal". The need for a comparative collection was addressed by the establishment of the Comparative Plant Fiber Collection (CPFC) at The Ohio State University. Supported by the National Science Foundation, the collection continues to expand and provides new insights into fiber use in prehistoric textiles. The Comparative Plant Fiber Collection The Comparative Plant Fiber Collection was established to provide comparative fibrous material processed from plant stems typical of those employed by prehistoric native Americans of eastern North America. The scope of the collection was limited, at first, to the examination of the fiber products that are very fine, long, and strong and can be used to produce finely twined textiles. Coarse yarns and cords that would result in coarse bags or other textiles and wood splits or entire plant pieces employed in basketry or mats were not originally included in the collection. Dyeplants were not included in the original scope of the CPFC but these are being added. Plant stems were collectedfromtwo geographic areas: southern and central Ohio in one area and northern Georgia. One set of the stems were processed within 5 days of collection in the field; the other set was allowed to dry in a desiccator for 6 weeks prior to processing. The fibers were processed from the plant stems in four ways: (1) hammering the stems, then hand peeling the individual fibers or fiber bundles; (2) soaking the stems in water for 2 weeks to simulate the effects of retting, then hand peeling the fibers and fiber bundles from the stems; (3) boiling the stems in demineralized water for 6 hours, allowing them to cool, then hand peeling the fibers from the stems; and (4) boiling the stems in demineralized water with potassium carbonate for 6 hours, and hand peeling the fibers from the stems. The behavior of the fiber bundles in processing was recorded. The fiber products have been studied by multiple techniques of optical microscopy, scanning electron microscopy, X-ray microanalysis, and infrared microspectroscopy. Details of the establishment of the collection were reported previously (77). The results obtained from these first analyses led to

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the categorization of the plant fibers based on polythetic groups of attributes (72). Behavior of the fiber bundles in processing provides information concerning the likelihood of use of certain plant fibers in finely twined textiles; only certain plant stems yield fine, strong, and long bundles which can readily be manipulated into fine yarns. Observation of morphology through the optical microscope also provides more attribute clues which aid classification. Each of the four groups proposed contains a number of plants whose fibers display generally similar characteristics. Group I plants are those which yield fine, strong, and long fiber bundles which are pliable enough to be twisted or spun into fine yarns. These bundles are easy to process from the plant stems and can be withdrawn in long strands without breaking. The fibers in these bundles are relatively clean-surfaced with periodic dislocations commonly attributed to bast fibers. Fioers of the genus Apocynum constitute a subgroup of the Group I fibers because they display characteristic surface folds. Fibers in the other three categories include hard-to-process fibers which do not yield fiber bundles of long lengths or fine diameter. Group II and III fibers display no distinctive surface characteristics. Fibers in Group IV are readily distinguished because of the presence of extensive quantities of crystal inclusions. More detailed descriptions of the group classifications are reported by this author (72) elsewhere. This manuscript reports the results of scanning electron microscopic examination and X-ray microanalysis of some of the CPFCfibers;these analyses revealed features unseen in optical microscopy. Also reported are the results of the study of high temperature "carbonized" and cold plasma ashed fiber products. Thefindingsreported herein will ultimately contribute to the refinement of the Group classifications. Experimental Methods A Zeiss Axioplan research microscope was employed in the optical microscopic examination of the fiber products. Scanning electron microscopic (SEM) examination was carried out employing a Jeol JSM-820 scanning electron microscope and X-ray microanalysis was accomplished with a Link Analytical eXL energy dispersive x-ray analyzer. Fibers were mounted on carbon planchettes and carbon coated for SEM-energy dispersive spectrometric (EDS) analysis. Fibers held in open ceramic dishes to allow contact with air and fibers wrapped in foil to eliminate air were "carbonized" in a muffle furnace at 600 °C for 5, 10, and 15 minutes. Fibers also were ashed in a SPI Supplies Plasma-Prep II plasma etching unit with oxygen gas plasma (75).

In Archaeological Chemistry; Orna, M.; ACS Symposium Series; American Chemical Society: Washington, DC, 1996.

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Results and Discussion Fiber Morphology. Various aspects of fiber morphology were investigated in this continuation of the study of the CPFC. Tables I and II summarize the findings of the SEM and EDS examination of a selection of fibers from Groups I, III, and IV. The possibility of alteration in morphology resulting from the four processing treatments was explored. Butterfly weed (Asclepias tuberosa), spreading dogbane (Apocynum androsaemifolium), and Indian hemp (Apocynum cannabinum) fibers which had been processed from the plant stems by the four treatments were studied with SEM and EDS. The Indian hemp fiber bundles displayed little difference as a consequence of the water soaking or water boiling treatments. In fact, the fiber bundles still possessed attached plant cellular materials. The fibers themselves displayed the surface folds which are characteristic of their genus. The Indian hemp fibers which had been boiled with potassium carbonate appear cleaner than fiber bundles resulting from the other three treatments. The nodes are prominent and the fibrils are apparent within the fiber. There is less attached intercellular material, although particles of material are observed on the fiber surfaces. Some of the particles contain sodium, potassium, and chlorine while others are carbonaceous. While one would attribute the presence of potassium to the potassium carbonate solution, it should be noted that potassium, chlorine, and sodium were found in the same fibers without the boiling treatment. Butterfly weed fibers also display localized potassium on the surfaces; after boiling with potassium carbonate the fiber surfaces are covered with bumps which have a high potassium and chlorine content. No attempt was made in this exploratory work to quantify the elemental composition of the fibers. After the water soaking treatment, the spreading dogbane fibers seem somewhat cleaner than those which had been processed by hammering and peeling. In addition, the fibrils are apparent in the fiber surfaces. Water boiled fibers appear clean only in some areas while in others cellular material like parenchyma cells remain. Even after treatment 4, boiling in potassium carbonate, the dogbane fibers still possess areas occluded by agglomerated cells. The evidence obtained indicates that even the boiling treatments are insufficient to remove distinctive surface characteristics such as dislocations or surface folds. Distinction between Group I and IV categories is based primarily on the presence of the profuse amounts of crystal inclusions found associated with the Group IV fibers. The question then arises whether the fiber morphologies alone would allow their identification if it were possible to eliminate the associated crystals and plant cells by some sort of extensive

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Table I. Morphology and Elemental Analyses of Selected Group I Plant Fibers

Other Features

Dislocation*

Surface Transverse Surface Mark Folds

Butterflyweed (OH)*

yes

no

no

Swamp Milkweed (OH)

yes

no

no

Common Milkweed (OH)

yes

no

no

Red Mulberry (OH)

yes

no

no

no

yes

some smooth, some with surface folds

yes

Parenchyma

Spreading Dogbane (OH)

Spreading Dogbane (GA)

Elemental Composition

Indian Hemp (OH)

yes

no

yes

covering occludes folds, some fibers with folds, some without

c,o

Intermediate Dogbane (OH)

yes

no

no

cambium occludes surface folds

C,0,small C1,K

Blue Dogbane (OH)

yes

no

no

smooth fibers

C,0,C1,K small Ca

Stinging Nettle (OH)

yes

yes

no

Parenchyma cell residue

C,0, Small Κ

Wood Nettle (GA)

no

yes

no

Parenchyma cell residue, logitudinal striations

False Nettle (OH)

no

yes

yes

longitudinal striations

*Each fiber common name is followed by the collection location.

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Table II. Morphology and Elemental Analyses of Selected Group ΙΠ and IV Plant Fibers

Dislocations

Transverse Surface Folds Surface Marks

Other Features

Elemental. Composition

Red Cedar (GA) *

smooth fibers

C,0, small Ca

Paw Paw (OH)

crystals embedded and covered

Black Walnut (OH)

smooth fibers

Black Willow (GA)

some surface disruption

Slippery Elm (OH)

smooth fibers

Each fiber common name is followed by the collection location.

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retting technique. Further study of the Group IV fibers was conducted to answer this question. While treatment 4 is not sufficient to completely clean the fibers in Group IV, fibers which had been boiled in potassium carbonate did reveal some areas which were free from crystal inclusions and could be examined. Of these, only two of the Group IV fibers, black willow (Salix nigra) and slippery elm (Ulmus fulva), appear to possess dislocations in any way similar to those seen in Group I fibers when examined with optical microscopy. Other fibers from Group IV which were examined did not appear comparable to Group I fibers. Basswood (Tilia americana), black walnut (Juglans nigra), and paw paw (Asminia triloba) fibers are smoothsurfaced fibers underneath their crystal and plant cell coverings. Further examination of black willow and slippery elm fibers with scanning electron microscopy revealed surfaces which are very distinctly different from those of the Group I fibers. Figure 1 displays the features of a typical Group I fiber, common milkweed; Figure 2 displays the structures observed in black willow fibers. There is some evidence for "ridges" (14) in these fibers which could have been misconstrued in optical microscopy as "dislocations". It has already been noted that certain fibers display surface folds which result in a unique appearance when examined with the optical microscope. The surface folds causefrequentsurface disruption. This unique characteristic has been pronounced as a key indicator of a particular genus of plant fibers, Apocynum (11,12). Further examination of fibers within this group supported this finding. Blue dogbane, though belonging to the Family Apocynaceae, is from the genus Amsonia, and does not display the surface folds. Thus, the distinction made between plant fibers is valid at the general level. Fiber samples from the same genus and species of plant but collected in different locations (Ohio and Georgia) or at different times (1991 and 1993) were compared for morphological differences. Fiber morphology and elemental composition are summarized in Tables I and II; shapes and elemental composition of inorganic inclusions and other features are summarized in Tables III and IV. Fibers from the same genus and species displayed consistent features despite the differences in circumstances of collection. Both examples of common milkweed (Asclepias syriaca), and red mulberry (Morus rubra) display the surface characteristics of Group I fibers. Calcareous druses, spherical clusters formed by accumulation of crystals, were found in both samples of common milkweed. Both examples of spreading dogbane (A. androsaemifolium), and Indian hemp (A. cannabinum) exhibit surface folds. Both examples of stinging nettle (Urtica dioica) and false nettle (Boehmeria cylindrica) display transverse markings. The crystal inclusions observed in basswood (T. americana), paw paw (Asiminia triloba), and black willow (Salix nigra) are consistent for materials collected at

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Figure 1. Common milkweed: Typical Group I morphology.

Figure 2. Black willow surface morphology.

In Archaeological Chemistry; Orna, M.; ACS Symposium Series; American Chemical Society: Washington, DC, 1996.

In Archaeological Chemistry; Orna, M.; ACS Symposium Series; American Chemical Society: Washington, DC, 1996. random few random few-infjlling

lump, not flat faced in lines between fibers

plate-like druse-like rectangular

Spreading Dogbane (GA)

Indian Hemp (OH)

parenchyma

lump, not flat faced in lines between fibers

druse-like particle druse-like crystals

Red Mulberry (GA)

Ca,P,0, small C , K Ca,P,0, small C , K Ca,0

S,K,C,0, small Si C,0,Mg,Si,S,K, small Ca

fluffy deposits sand globules

lumps fluffy materials fluffy materials

random

druse

Common Milkweed (OH) Ca,0,C

particles

Ca,Mg,0,C

random

random particle lumps on surface

plates, some beginning to form druse

Ca,0,C, small S,K, K.S, small Ca,Cl,C,0

Description

Common Milkweed (OH)

in strings between fibers

Elemental Composition

particles

druse particle/druse

Location

Swamp Milkweed (OH)

Butterflyweed (OH)*

Shape

Large Inclusion

Elemental Composition

Ca,P,K.O, small ΑΙ,Ο,Κ Κ,Ο, small Cl, Ρ

Si.O.K, small C,Mg,Cl,S Ca.O, small K,S K.C1.0, small P,Mg,S

Fe, Small K,0,S

Si,0,C

Al,Si,S,K,C,0, Si,0

Other Inclusion

Table ED. Morphology and Elemental Analyses of Inorganic Inclusions in Group I Plant Fibers

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In Archaeological Chemistry; Orna, M.; ACS Symposium Series; American Chemical Society: Washington, DC, 1996. in string

m string

* Eachfibercommon name is followed by the collection location.

False Nettle (GA)

Wood Nettle (GA)

plate-like particles

druse

Stinging Nettle (OH)

Stinging Nettle (GA)

lumps alongfibersurface

Blue Dogbane (OH)

Si,0, small S,K,Ca,C

K,Ca,S K,0,P,S,Ca small Mg some Si,0,K, S, small A1,P,C

random particles particles within parenchyma

Si.O, small A1,C

Si.O Ca

particles

particles in parenchyma

inorganic coatingfibersin some areas small lumps prevalent

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Table IV. Morphology and Elemental Analyses of Inorganic Inclusion in Group I Plant Fibers

other Inclusion

Large Inclusion

Elemental Composition

Shape

Location

Description Elemental Composition Particles

Red Cedar (GA)*

irregular, some particles, some blocks

distributed over surface

Ca,0

Paw Paw (OH)

longflatblocks with rounded ends druses

fill in parenchyma cells random

Ca, small 0

Paw Paw (GA)

long rectangular blocks, otherflatfaced shapes

in lines

Ca,0

small articles

Si

Ca,0

small particles

Si,Al

truncated bipyramids druse

Ca.O

small particles

Si

Black Walnut (OH)

Ca,0

Black Willow (OH)

truncated bipyramids

within each parenchyma cell

Ca.O

fluffy deposits Ca,0, Small Si

Basswood (GA)

long double pointed twinned

in lines along fiber

Ca,0

flattened lump Si,0, small K, Mg,Al

flat crystals druse

between fiber bundles

Ca,0

Basswood (OH)

double pointed twinned crystals

Ca,0,C

particles

Ca,S,K,C,0

* Eachfibercommon name is followed by the collection location.

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different locations and different times. The morphological consistencies further justify the categorization system proposed. In fact, the size, shape and chemistry of the formations resulting from calcification processes are "under close cellular control" (75, 16) and thus may prove to be class specific indicators. In a similar manner, the process of silicification is specific and the presence of opal phytoliths have been proposed as a route to determining use of plants in prehistoric societies (77). Red cedar (Juniperus virginiana) fibers have been placed in Group III; Jakes, Chen, and Sibley (72) indicate that no inorganic inclusions were observed in optical microscopic study of red cedar fibers. Subsequent scanning electron microscopic examination of thesefibers,however, reveal small calcareous inclusions of irregular shape distributed over the fiber surfaces: The inclusions are not located in a specific region as are crystals noted in other fibers. The inclusions, shown in Figure 3, are 1-3 μπι in size, much smaller than the crystals observed in the Group IV fibers which can reach 20 μιη in size. Stinging nettle (U. dioica), false nettle (Boehmeria cylindrica), and wood nettle (U. divaricatum) were examined under the SEM. These nettles display longitudinal striations and periodic transverse surface marks (Figure 4). It is possible that these marks are linked to the presence of residual parenchyma cells. The feature appears to be characteristic of the Urticaceae family but no further distinction according to genera can be determined. Carbonized fibers. In the original design of the CPFC research, a high temperature carbonizing treatment offiberswas proposed to experimentally replicate the "charred" or "carbonized" fibers found in many prehistoric native American textiles. The terms are used interchangeably in the literature, although technically it is unlikely that the charred fibers have, in fact, been reduced completely to carbon alone. Fiber products produced through the four treatments were subsequently heated at 600 °C in air and without air (nominally) wrapped in foil. Observation of these black materials by reflected light microscopy was attempted. Extensive work in the examination of carbonized Indian hemp fibers has been reported by Srinivasan (18). The microscopist can make some observations as the fibers are scanned under the microscope, but the field of view is very irregular in all cases, precluding the collection of useful pictures. Scanning electron microscopic study of these carbonized products is necessary to accurately describe the alterations in structure which occur as a consequence of high temperature carbonization. Srinivasan (18) also improved the design of the high temperature experiments. Since the foil wrapping did not eliminate oxygen access entirely but did inhibitfibermotion during heating, she carbonized Indian

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Figure 3. Eastern red cedar inclusions.

Figure 4. Stinging nettle: Transverse cross markings.

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hemp fibers held in open crucibles in an inert argon atmosphere. The fibers coiled and shrank and some residual marks remained which are possibly related to the location of the dislocations. It is apparentfromthese experiments that much more extensive work is required to adequately describe the change in structure of carbonized fibers which results from high temperature carbonization. Consequent application of suchfindingsto the identification of carbonizedfibersin prehistoric textiles also will require extensive research. Cold Plasma Ashed Fibers. In examination of the carbonized fibers prepared in this research, questions arose concerning the inorganic inclusions revealed through high temperature incineration of the organic matrices in which they are supported. In fact, the carbonization experiments were comparable to some of the methods employed in phytolith research in dietary plants (19-21). After further exploration of the methods employed in phytolith preparation and the pitfalls of each, a cold plasma ashing method for phytolith preparation was developed (75) which avoided the consequences of high temperature incineration and of strong acid digestion. Cold plasma ashing has been shown to be a promising method for the preparation of phytolith samples from plant material. Fibers can be ashed in place on a carbon planchette, thus allowing the observation of the exposed inorganic inclusions embedded in the skeleton of remaining organic material. The relationship of the location of these crystals and inclusions with other plant cells can be observed. While the larger crystalline inclusions present in Group IV fibers were observed microscopically even prior to ashing, the ashing procedure produces clean-surfaced crystals unobscured by tissue and reveals other less obvious inclusions. An additional advantage of the procedure is that since the ashed fibers are prepared for scanning electron microscopic examination, they can also be analyzed for elemental composition through energy dispersive analysis of X-rays. The morphological and elemental data on inorganic inclusions obtained through the study of cold plasma ashed fibers from the CPFC provides new clues to enhance the Group classification scheme. The Group IVfibersyield many types of crystal inclusions (Table IV). Basswood (Γ. americana) (both from Ohio and Georgia) possesses both siliceous particles and large (10-20 μπι) double-pointed twinned calcareous crystals (Figure 5). The crystals are aligned along the phloemfibersbut do not appear to form within the parenchyma cells as do the twinned calcareous crystals observed in paw paw (Asiminia triloba)fibers.These flat-faced crystal blocks (Figure 6) are rounded on the ends and appear to have grown to fit within the parenchyma cell. Black willow and black walnut display calcareous truncated bipyramids (Figure 7) as well as some small siliceous particles. The bipyramids can be seen within the walls of the parenchyma cells. The brick-like rectangular

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Figure 5. Basswood: Twinned double pointed crystal inclusions.

Figure 6. Paw paw: Twinned rounded plate crystal inclusions.

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blocks observed in slippery elm (Ulmus fulva) also align with the phloem fiber cells, but are not formed inside parenchyma cells. The calcareous crystals of the woody Group IV fibers are so predominant that they obscure the much smaller siliceous particles that are present. While further work is necessary, it appears that the crystal shape and composition of the woody Group IV phytoliths may well prove to be a useful attribute in distinction among members of that Group. Although observed in SEM in fibers which were not ashed, the small inorganic inclusions covering the surface of red cedar fibers are more easily studied in plasma ashed material. Figure 3 displays the particles revealed after ashing the fiber bundles. While EDS analyses of these inorganic inclusions revealed elemental compositions of calcium and oxygen or silicon and oxygen, the form of these materials was not evaluated. It is likely that the compounds represented are calcium oxalate and silica (75, 16, 19, 20, 21). Optical microscopic examination of Group I fibers reveals that some possess associated inorganic structures. By reducing the organic matrix, plasma ashing of these materials exposes the crystals and other inclusions so that they can be more readily observed. In effect, the inclusions which may be random and infrequent are "concentrated" by the ashing process, as the organic composition of the plant material is reduced and their elemental composition can provide information as well. The Group I fibers display many structures of both calcareous and siliceous composition (Table III). Druse crystals (Figure 8) are observed in milkweed fibers (A. tuberosa, A. incarnata, A. syriaca) and in red mulberry fibers ( M rubra), in agreement with the optical microscopic observations. These druses are large and predominantly calcium in composition. Other calcareous plates and particles are observed as well as rounded and irregularly shaped siliceous particles. Common milkweed fibers also display fluffy globular surface structures which, in some areas, have a high potassium content. Swamp milkweed displayed large siliceous faceted inclusions. Druses were not observed in the dogbanes or Indian hemp fibers. Indian hemp, in one instance, did display an agglomeration of plate-like crystals which could possibly be called a druse. More common in the Indian hemp fibers are plate-like crystals which form within cell walls (Figure 9). Indian hemp also displays fluffy looking globular structures with a high potassium, phosphorous, and calcium content. Calcareous druses were observed in stinging nettle fibers, while plates were seen in wood nettle fibers. The stinging nettle also displayed a siliceous coating of the fibers in some areas (Figure 10) and small siliceous lumps

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Figure 7. Black willow: Truncated bipyramid crystal inclusions.

Figure 8. Common milkweed: Druse crystal inclusion.

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Figure 9. Indian hemp: Block crystal inclusions.

Figure 10. Stinging nettle: Siliceous coating over fibers.

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Figure 11. Stinging nettle: Siliceous particle within a cell.

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were observed within the cell walls (Figure 11). The number of any of these inclusions is small in comparison to the number seen in Group IVfibersand their presence alone is not considered diagnostic for the plant fiber genera but their presence may provide an additional clue useful in plant fiber indication and group classification of unknown plant fibers. Cold plasma ashing reveals structure previously occluded by plant cellular structures and so is useful in developing an atlas of the plant fiber inorganic inclusions.

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Conclusion Continued work conducted in the study of fibers from the CPFC has provided a variety of insights. The intrinsic morphology offibersappears to be maintained despite the processing treatment employed. Thus enough of the characteristic features of the plant fibers are maintained in processing that the fibers can be identified by the group classification scheme. The surface features of the Group IVfibersalone are not enough to provide categorization but they are distinct from the characteristic dislocations observed in Group I fibers. Carbonization of fibers still needs much work to provide useful information leading to the identification of charred archaeological fibers. Cold plasma ashing of the plant fibers is a useful method for the preparation and observation of inorganic inclusions in fibers. The Group IV phytoliths and other inclusions are distinct and are multitudinous, and so may aid in subclassification within the group. Group I phytoliths and other inclusions are less prevalent than those observed in Group IV and may only provide a useful clue to identification of fibers. Acknowledgments The award of the National Science Foundation Grant BNS-9021275 provided the initial support for this work. Additional salaries and support were provided in part by state and local funds appropriated to the Ohio Agricultural Research and Development Center, The Ohio State University. Dedication This paper is dedicated to the memory of Lucy R. Sibley. Just as the lives of the peoples of the past live on through the textiles they left behind, the life of Lucy R. Sibley will continue through her contributions to this field of research. Literature Cited 1. Willoughby, C. C. Ohio Arch andHist.Quarterly 1938, 47, 273-287. 2. Whitford, A. C. Anthropological Papers of the Museum of Natural History 1941, 38, 5-21.

In Archaeological Chemistry; Orna, M.; ACS Symposium Series; American Chemical Society: Washington, DC, 1996.

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