BIOMEDICAL AND ENVIRONMENTAL APPLICATIONS OF SECONDARY ION EMISSION MICROANALYSIS

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BIOMEDICAL AND ENVIRONMENTAL

APPLICATIONS OF SECONDARY ION EMISSION MICROANALYSIS

R. Linton

To cite this version:

R. Linton. BIOMEDICAL AND ENVIRONMENTAL APPLICATIONS OF SECONDARY ION EMISSION MICROANALYSIS. Journal de Physique Colloques, 1984, 45 (C2), pp.C2-553-C2-555.

�10.1051/jphyscol:19842126�. �jpa-00223795�

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JOURNAL DE PHYSIQUE

Colloque C2, supplgment a u n02, Tome 45, f g v r i e r 1984 page C2-553

BIOMEDICAL AND ENVIRONMENTAL APPLICATIONS OF SECONDARY ION EMISSION MICROANALYSIS

R.W. L i n t o n

Department of Chemistry, University of North CaroZina, Chapel Hill, NC 27514, U.S.A.

Resume

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Le SIMS e s t a p p l i q u e 1 l a c a r a c t e r i s a t i o n de c o n t a m i n a t i o n s dues au m i l i e u ambiant e t 1 l ' e t u d e c y t o p a t h o l o g i q u e de p a r t i c u l e s r e s p i r a b l e s dans l e s macrophages a l v e o l a i r e s .

La q u a n t i f i c a t i o n des donnees SIMS p r o v e n a n t d ' C c h a n t i l l o n s b i o l o g i q u e s e s t en p a r t i e l i m i t e e p a r des i r r @ g u l a r i t @ s de s u r f a c e dues au rFitr@cissement d i f f e r e n t i e l de l ' e c h a n t i l l o n p e n d a n t s a d e s h y d r a t a t i o n e t p a r une v a p o r i s a - t i o n heterogene o u d i f f e r e n t i e l l e des d i v e r s c o n s t i t u a n t s t i s s u l a i r e s . Dans l e s etudes p r e l i m i n a i r e s on a compare l e s e f f e t s morphologiques du decapage p a r plasma ou p a r bombardement i o n i q u e s u r d i f f e r e n t e s p r 6 p a r a t i o n s .

A b s t r a c t

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The a p p l i c a t i o n o f S I M S t o t h e c h a r a c t e r i z a t i o n of environmental contaminants i s described, i n c l u d i n g t h e s t u d y o f t h e c y t o p a t h o l o g y o f r e s p i r a b l e p a r t i c l e s i n a l v e o l a r macrophages.

Q u a n t i f i c a t i o n o f S I M S d a t a o f b i o l o g i c a l specimens i s l i m i t e d i n p a r t by s u r f a c e topography i n t r o d u c e d by d i f f e r e n t i a l s h r i n k a g e d u r i n g specimen d e h y d r a t i o n and by non-uniform o r d i f f e r e n t i a1 s p u t t e r i n g o f v a r i o u s t i s s u e components. P r e l i m i n a r y s t u d i e s a r e discussed comparing t h e morphologic e f f e c t s of r a d i o f r e q u e n c y plasma e t c h i n g vs. i o n beam s p u t t e r i n g on v a r i o u s t i s s u e p r e p a r a t i o n s .

The development o f a n a l y t i c a l t e c h n i q u e s f o r t h e morphochemical c h a r a c t e r i z a t i o n o f t r a c e and/or t o x i c elements i n v a r i o u s e n v i r o n m e n t a l o r b i o l o g i c a l media i s a c u r r e n t s u b j e c t of i n t e r d i s c i p l i n a r y research. The elemental coverage, d e t e c t i o n s e n s i t i v i t y , and s p a t i a l r e s o l u t i o n o f secondary i o n emission m i c r o a n a l y s i s (SIM) o f f e r t h e promise o f unique a n a l y t i c a l c a p a b i l i t i e s i n t h i s a p p l i c a t i o n s area (1,2). Our c u r r e n t c o l l a b o r a t i v e SIM s t u d i e s i n v o l v i n g t r a c e o r t o x i c element c h a r a c t e r i z a t i o n i n b i o l o g i c a l specimens i n c l u d e : l u n g t i s s u e p a t h o l o g y , a t h e r o s c l e r o s i s , neuronal t i s s u e s impacted by A l z h e i m e r ' s disease, and i n v i t r o s t u d i e s o f c e l l mono1 ayers.

One s p e c i f i c s u b j e c t under i n v e s t i g a t i o n i n o u r l a b o r a t o r y has been t h e SIM s t u d y of t h e surface c h e m i s t r y of v a r i o u s a i r b o r n e p o l l u t a n t p a r t i c l e s . A t i s s u e i s t h e r o l e of t h e p a r t i c l e s u r f a c e i n m e d i a t i n g b o t h heterogeneous atmospheric r e a c t i o n s and p a r t i c l e t o x i c i t y subsequent t o d e p o s i t i o n v i a i n h a l a t i o n o r i n g e s t i o n . P a r t i c l e s produced i n h i g h t e m p e r a t u r e i n d u s t r i a l processes f r e q u e n t l y e x h i b i t s u r f a c e enrichments o f sorbed i n o r g a n i c and o r g a n i c compounds ( 2 ) . A few examples o f SIM r e s u l t s f o r a u t h e n t i c p o l l u t a n t m a t e r i a l s a r e l i s t e d i n Table 1.

To i n v e s t i g a t e t h e r o l e of surface e n r i c h e d components on p a r t i c l e t o x i c i t y , experiments have been i n i t i a t e d i n v o l v i n g i n v i t r o s t u d i e s of p a r t i c l e i n t e r a c t i o n s w i t h a l v e o l a r macrophages. ~ e n e r n c h n i q u e s i n v o l v i n g sample p r e p a r a t i o n and c o r r e l a t i v e m i c r o s c o p i c e x a m i n a t i o n have been d e s c r i b e d p r e v i o u s l y (3-5). The d i s t r i b u t i o n of phagocytosed Pb from v a r i o u s l e a d o x i d e c o a t e d p a r t i c l e s ( i n c l u d i n g c o a l f l y ash) has been

Article published online by EDP Sciences and available at http://dx.doi.org/10.1051/jphyscol:19842126

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C2-554 JOURNAL DE PHYSIQUE

T a b l e

1.

SIM o f t h e S u r f a c e Layers o f A i r b o r n e P a r t i c u l a t e P o l l u t a n t s .

-

C o n c e n t r a t i o n i a t i o s ( S u r f ace/Bul k ) Element

C r

Coal F l y Automobi 1 e S t e e l B l a s t

Ash Exhaust Furnace Dust

3.3

- ^T-

a. secondary i o n i n t e n s i t y change observed w h i l e s p u t t e r i n g ⁺ a p p r o x i m a t e l y t h e o u t e r lOOOA of t h e p a r t i c l e surface w i t h O2

.

b. F e (as i r o n o x i d e ) i s a p r i n c i p a l b u l k o r m a t r i x component o f a l l t h r e e p a r t i c l e types.

e s t a b l i s h e d u s i n g TEM i n c o n j u n c t i o n w i t h e l e c t r o n and i o n m i c r o p r o b e a n a l y s i s (4, 5 ) . D i s s o l u t i o n and m i g r a t i o n o f Pb f r o m t h e s u r f a c e s o f phagocytosed p a r t i c l e s r e s u l t e d i n t h e l i b e r a t i o n o f i n t r a c e l l u l a r Pb, r e p r e c i p i t a t i o n o f Pb as P and Ca

-

c o n t a i n i n g complexes t h r o u g h o u t t h e c e l l , and c y t o p a t h i c changes i n c l u d i n g s w e l l i n g o f t h e m i t o c h o n d r i a and n u c l e a r membranes.

One l i m i t i n g f a c t o r i n t h e q u a n t i t a t i v e i n t e r p r e t a t i o n o f SIM d a t a i n t h e above s t u d y was t h e apparent non-uniform o r d i f f e r e n t i a l s p u t t e r i n g o f v a r i o u s c e l l components ( 6 ) . T h i s i s a phenomenon of general concern i n t h e SIM c h a r a c t e r i z a t i o n of b i o l o g i c a l specimens. For a l v e o l a r macrophages, secondary e l e c t r o n images ( F i g . IA, 1B) suggest t h a t t h e n u c l e o l u s , n u c l e a r envelope and plasma qembrane a r e more r e s i s t a n t t o s p u t t e r removal b y t h e p r i m a r y i o n beam (02 1. However, these f e a t u r e s a l s o a r e observed t o

p r o t r u d e f r o m t h e surfaces o f u n s p u t t e r e d s e c t i o n s o f p e l l e t i z e d c e l l s ( 7 ) as

F i g u r e 1. Secondary e l e c t r o n images o f C coated r a b b i t a l v e o l a r macrophages ( w i dth=15pm).

A ) Snap f r o z e n , f r e e ~ e ~ d r i e d c e l l B) C e l l a f t e r lOkeV O2 bombardment

C ) Snap-frozen, f r e e z e - d r i e d 0.2pm t h i c k s e c t i o n o f p e l l e t i z e d c e l l s . S i n g l e Arrow = n u c l e o l u s ; Double Arrow = n u c l e a r envelope

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t h e apparent consequence o f d i f f e r e n t i a l s h r i n k a g e processes d u r i n g d r y i n g ( F i g u r e 1C). Such s t r u c t u r e s , t h e r e f o r e , may have i n h e r e n t l y g r e a t e r mass, d e n s i t y and t h i c k n e s s i n t h e dehydrated t i s s u e . Consequently, t h e

i n t e r p r e t a t i o n o f apparent s p u t t e r i n g a r t i f a c t s i s c o m p l i c a t e d b y t h e e f f e c t s of sample p r e p a r a t i o n . F u t u r e c o l d s t a g e SIM on f r o z e n , h y d r a t e d t i s s u e s may be a c o n s i d e r a b l e advantage i n l i g h t o f these p o t e n t i a l s p u t t e r i n g o r

p r e p a r a t i v e a r t i f a c t s .

A p r e l i m i n a r y c o m p l i l a t i o n o f t h e v a r i o u s e f f e c t s o f i o n s p u t t e r i n g o r r a d i o f r e q u e n c y plasma e t c h i n g on t i s s u e morphology i s shown i n Table 11.

Table 11. E t c h i n g E f f e c t s on B i o l o g i c a l Tissues.

Source

,

^A

T i s s u e Component 0 rf plasma

---

2keV A r ' beam

-

lOkeV

e2'

beam

+

m r heterochromat i n

-' ^- ^- ^-

Nuclear envelope

+

N u c l e o l us

+

L i p i d d r o p l e t

+

1 ysosomes

+

Z l i n e s ( s k e l e t a l muscle)

+

M l i n e s ( s k e l e t a l muscle)

+ +

= p r e s e r v e d d u r i n g e t c h i n g

-

⁼p r e f e r e n t i a l l y etched ND = n o t determined

The r e s u l t s o f ~ r + and 0 ⁺i o n beam bombardment a r e comparable s u g g e s t i n g t h a t s p u t t e r i n g processeg r e f l e c t c o l l i s i o n a l momentum t r a n s f e r l a r g e l y independent o f p r i m a r y beam composition. The chemical " m i c r o i n c i n e r a t i o n "

induced b y t h e O2 rf plasma, however, appears t o have a + s u b s t a n t i a l l y d i f f e r e n t s e l e c t i v e e t c h i n g b e h a v i o r r e l a t i v e t o t h e O2 i o n beam.

References

1. GALLE, P., LEFEVRE, R., M i c r o s c o p i c a Acta, Suppl. 2: M i c r o p r o b e A n a l y s i s i n B i o l o g y and Medicine, 1978, 341-354.

2.

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LINTON<R. W ~ H A R V E Y , D. T., CABANISS, G. E., "Environmental A p p l i c a t i o n s o f S u r f a c e A n a l y s i s Techniques", i n A n a l y t i c a l Aspects o f Environmental Chemistry, D. Natusch and P. K. Hopke, eds., ~ o h n y a n d Sons, New York, 1983, pp. 137-189.

3. MASTERS, S. K., e t al., SEM/1979/III, pp. 97-122.

4. LINTON, R. W., e t al., SEM/1980/II, pp. 583-596.

5. DEVRIES, C. R., e t al., Lab. I n v e s t .

48

(1983) 35-44.

6. LINTON, R. W., e t al., SEM/1982/III, pp. 1191-1204.