5 INTRODUCTION Hair samples are good resource biomaterials in forensic science and physical anthropology because various chemicalsand biological substances that accumulate in hair can bedetected and measured.Environmental factors including sunlight, air pollution , wind , salt water and fungiMany hair samples taken from sites of archaeological orforensic investigation have undergone weathering for longperiods of time.An ultramicroscopical study on the morphologicalchanges that occur in weathering hair samples would beuseful.
6 Materials and Method Ten hair samples 20–25 cm long Hair samples weathered for 5–25 yearsTombs in Kangneung,Korea.Males female,aged 50–60 years old.Five to 10 hair samplesSamples 3 to 5 tombs of individuals buried for 5, 10, 15, 20, and 25 years.Scanning electron microscope (SEM)Transmission electron microscope (TEM)
7 TEMHair samples were immersed in 2.5% paraformaldehydeGlutaraldehyde in neutral 0.1 M phosphate buffer for 1 h.Tissues were postfixed for 1 h in 1% (w/v) osmic acidDehydrated in graded ethanol,
8 SEM Hair samples were prefixed by immersion in 4% paraformaldehyde 0.1% glutaraldehyde in neutral 0.1 MPost-fixed for 2 h in 1% (w/v) osmic acidSamples were treated in a graded ethanolseries and isoamyl aceteateDried in a critical point dryer
9 The middle portion of the hair shaft from a living individual. ResultsThe middle portion of the hair shaft from a living individual.(A) A cross section showing the cuticle(c) , cortex (Co) and medulla (M).Melanin pigment is observed in the cortex layer.(B) Scanning electron microscopy (SEM) showing well preserved cuticular scales.
10 clearly visible. Arrows indicate melanin pigment granules. (C) SEM of the cross section, with the cuticle (c), cortex (Co) and medulla (M)clearly visible. Arrows indicate melanin pigment granules.
11 (D) Transmission electron micrograph (TEM) of the cross section of a cuticle(E) Highly magnified TEM image of the cortex.
12 The terminal portion of the hair shaft from a living individual. (A) A cross section showing an irregular shape and a thinner cuticular layerthan in Fig. 1A.(B) SEM data showing loss of the cuticle layer and exposure of the underlying cortex.
13 (C) Magnified image of the SEM showing loss of the cuticle and exposure of the underlying cortex.
14 (D) TEM image of the cuticle and cortex layers (D) TEM image of the cuticle and cortex layers. Arrows indicate the loss of cuticle scales.
15 (A) and (B) Controls, showing that the general Histological changes of hair shafts during 25 years of weathering. (Left) Representative external morphology under the stereomicroscope. (Right) A representative toluidine blue stained cross section under the light microscope.(A) and (B) Controls, showing that the generalstructure of the middle portion of hair from a living individual is well preserved.
16 (C) and (D) Five years after death. Histological appearances similar to (A) and (B).(E) and (F) Fifteen years after death, showing rougher surface and partial detachment of the cuticle layers.
17 (I) and (J) Twenty-five years after death, showing a (G) and (H) Showing damage to most of the cuticle layer and exposure of the underlying cortex to the environment (white arrow in H).Weathering damage proceeded from the medullar to the cortical layer (black arrows in H).(I) and (J) Twenty-five years after death, showing ahole in the hair shaft (white arrow in J)
18 TEM and SEM images of samples 5, 10 and 15 years. (A) and (B) SEM of a sample 5 years after death, showing that the hairshaft and root (HR) appear intact. S, scales in the cuticle.
19 10 yrs.(C) and (D) SEM of a sample 10 years after death, showing that the scales (s) on thecuticle layer have started to detach from the underlying structures (circle).
20 15 yrs. (E) and (F) SEM of a sample 15 years after death, showing retention of the general structure of the hair shaft
21 20 yrs. (A) SEM image, showing an increase in the number and sizes of pores in the hair shaft 20 years after death.(B) SEM image, showing detachment of the entire cuticle layer and macrofibrils(C) SEM image, showing extension of the pores into the cortex and medulla of the hair
22 (E) Highly magnified TEM image, showing spaces 20 yrs.(E) Highly magnified TEM image, showing spacesaround the macrofibrils (MFs) in the cortex and the appearance of melanin pigment granules(D) TEM image of the hair shaft cross section.
23 showing that only the outer 25 yrs.(F) and (G)SEM 25 years after death,showing that only the outercortex remained
24 25 yrs. (H) TEM comparisons with younger samples, showing that the spacebetween the macrofibrils increasedand the cuticle layer haddisappeared.