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INDUCED SENSIBILIZED FLUORESCENCE OF NORMAL AND ATYPICAL CELLS AND TISSUES
N. Tankovich, E. Baranov, L. Osokina
To cite this version:
N. Tankovich, E. Baranov, L. Osokina. INDUCED SENSIBILIZED FLUORESCENCE OF NORMAL
AND ATYPICAL CELLS AND TISSUES. Journal de Physique Colloques, 1987, 48 (C7), pp.C7-283-
C7-285. �10.1051/jphyscol:1987763�. �jpa-00227066�
JOURNAL DE PHYSIQUE
Colloque C7, suppl6ment au n012, Tome 48, dkcembre 1987
INDUCED SENSIBILIZED FLUORESCENCE OF NORMAL AND ATYPICAL CELLS AND TISSUES
N.I. TANKOVICH, E.P. BARANOV and L.I. OSOKINA
I.V. Kurchatov Institute of Atomic Energy, Moscow, USSR
The problem of selective accumulation of fluorescent dyes by neoplastic fornations is actual and has not been solved so far. Classic luininescent histo- and cytochemistry employs vari- ous fluochromes,including well-known sodium fluorescein. This dye presents great interest due to its high fluorescence quantum output and increased accumulation by atypical cells in vitro et in vivo(I),
This article presents the studying of accumulation Of sodium fluorescein(F1Na) in cells, organs and tumours of mice employing fluorescent analysis. The experiments were car-ied out on mice series C Black and FI with replanted Erlichts tumour(so1id form) and the VUT-5. 57
A
number of series of experiments were carried out, with I0 mice used in each group. 0,2 I X ~ of PlNa were injected into retroorbital sinus('?,
5
mg/kg and 75 mg/kg of animal weight).The fluorescence intensity was evaluated in kidneys, liver, spleen, lungs and tumour 30 min., 1,2,3,4,6,12,24 hours after FlNa was injected.
Tbe characteristic form of spectrum and fluorescence inten- sity of the same objects without FlNa injection served as refe- rence sample.
The fluorescence spectra were registered by the fluorescence spectrophotometre at
1
ex=441 m, fluorescence intensity was measured at 1=520 nm.The pieces of tissues to be researched were placed into screen window, which pemitted to maintain the same standard dimensions of samples, Fluorescence intensity was expressed in relative units. The obtained data was processed according to the variati- onal statistics methods.
Article published online by EDP Sciences and available at http://dx.doi.org/10.1051/jphyscol:1987763
C7-284 JOURNAL DE PHYSIQUE
The r e s u l t s of r e s e e r c h e s shovred t h a t w i t h FlXa b e i n g i n j e c s - e d a t a d o s e of 7 , 5 m g / k ~ o r l e s s t h e lunlinescence of some tiss- u e s ( i n c l u d i n g t u x o u r s ) hapnened t o be comparable w i t h t h e tiss- u e s ' own c h a r a c t e r i s t i c f l u o r e s c e n c e i n t h e f l u o r e s c e i n s p e c t r w i . a r e c
.
The f i g . 1 s h o n s r e s u l t s of d e t e r m i n i n g of deperd-erce o f r e - l a t i v e f l u o r e s c e n c e i n t e n s i t y of v a r i o u s o r g a n s and E r l i c h ' s tumour upon tile t i ~ e of a n i m a l s
'
i n j e c t i o n w i t h 7 5 m&/kf; of FlNz, a s w e l l a s t h e a r e a of f l u o r e s c e n c e of o r g a n s t h e m s e l v e s and o f v a r i o u s t u n l o u r s ( I 0-
I 2 0 r e l . u n . 1 .The i n t e n s i t y of t h e t w - o u r f l u o r e s c e n c e exceeded t h a t o f t h e o r g a n s e x c e p t i n g kidney
-
o r g a n , t h a t t a k e s FlNa o u t of o r - ganism. Itidneg i s f o l l o w e d by t h e l i v e r , l u n g s , s p l e e n by t k e i r s t a g e o f dye a c c u m u l a t i o n . T n a t l s why i n t h e l a t e r e x 2 e r i n e n t s the s t a g e o f P l K a a c c u m u l a t i o n by t h e tumour was e v a l u a t e d i n comparison w i t h t h e l i v e r .The i n t e n s i t y of tumour f l u o r e s c e n c e i n I, 2 , air-d 3 h o u r s a f t e r t h e dye was i n j e c t e d exceeded t h a t of t h e l i v e r I 3 , I I and 9 t i n e s a c c o r d i ~ g l y .
The a n a l y s i s of s p e c t r a shows t.lat I h o u r a f t e r t h e F1Na i n - j e c t i o n i s a n o p t i m a l t i m e f o r o b s e r v a t i o n of dye s e l e c t i v e a c c u m u l a t i o n by n i c e
'
t m o u r s .S i m i l a r c h a r a c t e r of FlNa a c c u m u l a t i o n was n o t i c e d w i t h t h e XUT-5,a t m o u r o f d i f f e r e n t g e n e s i s compared w i t h t h e E r l i c h ' s , I n I h o u r a f t e r FlXa i n j e c t i o n t h e f l u o r e s c e c c e i n t e n s i t y o f t e VY'UT-5 and t h a t o f a n i m a l ' s l i v e r were: 4726 and 543 r e l . u n . r e s p e c t i v e l y t l - a t o f E r l i c h ' s tm.our and l i v e r
-
3376 and 240 r e l . un. r e s p e c t i v e l y .X v i d e n t l y there 's such a moment when t h e f l u o r e s c e n c e i n t e n - s i t y o f t m o u r f s meximun. L o s t p r o b a b l y t k a t t h e s e p a r a m e t e r s a r e r e l a t e d t o t h e t y p e o f tumour, i t s l o c a l i z a t i o n , c h a r a c t e r of t h e o r g a n i s m ' s metabolism, e t c . T h e r e ' s some i n f o r m a t i o n a b o u t t 3 e i m p o r t a n c e of membrane c o n t a c t s f s t a t e between c e l l s when FllJa i s b e i n g accumulated o r e x t r a c t e d by c e l l s : i n c a s e
s l o t c o n t a c t s a r e d e f e c t e d , t h e dye between c e l l s i s n o t t r a n s - g o r t e d ( 2 ) .
C e l l s u s p e n s i o n s i n c u l t u r a l medium were u s e d f o r t h e a n a l y s - i s of dye accumulation by a t y p i c a l c e l l s of t h e above-mentioned tuiaours. The c e l l s 1 c o n c e n t r a t i o n and t h e i r v i a b i l i t y were de- t e r n i r e d b e f o r e t h e e x p e r k n e n t s t a r t e d v:ith t h r i p a n e - b l u e dye.
PlNa s o l u t i o n was i n j e c t e d i n t o t h e mediuq w i t h c e l l s i n p o r t i o n o f 0 , I rnlcl of 0 , 0 5 $ - s o l u t i o n p e r 5C :r-5 of n e 2 i u r .
The cells vrere kept in themostate for 30 min. with further sedimentation at IOOC rev/min and were washed off the dye then with Chenxfs solution. The fluorescence intensity of the dye in cells in the area of about 520 nm was measured with photometric attaclvnent on luinescent microscope, with the He-Cd lazer irra- diation of =445,6 rm being an excitin~ light.
Ehen estirrating the cell fluorescence of various lines,it was discovered that each type of atypical cells is characterized by its own parameters of FlJJa accumulation an;d co~siderably exceeds sensibilized fluorescence of normal cells.
However, mechanism of selective accumulation of dye by aty- pical cells and tm~our tissue still doesn't present a comple- tely clear picture.
So we make a conclusion that at a dose of 75 mg/kg of FlNa iii I hour after mice were injected the tmours' fluorescence intensity becomes an order of magnitude as much as that of other organs(liver, lungs, spleen), which permits to differentiate the turcsour from tis~ues of other organs.
References
I.
N.G.Vinogradova, B.I.Tenkovich-
Dokl.Akad.MauL SSSR,v.285,2, p.459yI985.2. TJadeiL., Schindler TG.,
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Science, v. 232,4749, pp.525-526,1986,Dependence of reletive fluorescence intensity of various organs and Erlichfs tunour upon the time after FlNa injection to the animals: kidney(x1, turnour(@), liver ( 0 ), lung( 0 ),
