Khoa hoc Nong nghiep I Chan nudi DOI: 10.31276/VJST.64( 12).54-59 ? ? Anh luffing cua phirovg phap thuy tinh hoa va giai doan phat trien cua phoi den hieu qua bao quan lanh phoi bo san xuat in vitro tai Viet Nam f f Nguyen Khanh Van1*, Vu Thi Thu Huong1, Pham Thj Kim Yen1, Hoang Thi A u \ Pham Doan Lan2, Lira Quang Minh3 1P h o n g T hi nghiem diem cong nghe te bao d o n g vat, Vien C han nudi 2Vien Chan nudi 3B o K hoa hoc va C ong nghe N g a y n h an b 22/8/2022; n g a y c h u y e n p h a n b ie n 25/8/2022; n g a y n h an p h a n b ie n 26/9/2022; chap nhan d a n g 30/9/2022 Tom tat: lYluc dich cua nghien cun nham so sanh m ot so p h uang phap thuy tinh hoa va giai doan phat trien cua phoi bo san xuat in vitro den hieu qua bao quan lanh phoi bo in vitro tai V iet N am Phoi dirge dong Ianh theo p h u an g phap: (1) Thuy tinh hoa bang cryotop va (2) Thuy tinh hoa bang m icrodrop Ty le phoi bo thu dirge sau giai dong cua phirong phap (1) cao han so vd i (2) (100 so vdi 81,01% , p0,05) Trong nghien ciru nay, ty le hop tir song, phan chia, tao phoi nang va phoi nang thoat m ang sau giai dong tirong irng dat 86,83, 63,8 ,2 ,3 va 3,91% N ghien cun khong nhan thay s y khac biet ve ty le phoi song, phoi thoat m ang sau giai dong gitra nhom phoi dau/dau chat va phoi nang (tuwng ung 82,67 so v o i 88,43% ; 16,54 so v o i 20,34% , p>0,05) K it qua cho thay, phoi bo in vitro diro'c dong lanh cong & giai doan hop tir, phoi dau/dau chat hoac phoi nang bang phm m g phap thuy tinh hoa Tir khoa: cryotop, m icrodrop, phoi bo san xuat in vitro, thuy tinh hoa Chi so phan loai: 4.2 D$t vdn c a c h o a t d o n g c u a e m y m n o i b a o , h o h a p t e b a o , tr a o d o i in vitro-, b a o q u a n l a n h p h o i b o in v itro d a v a d a n g d u o c s u Q u y t r in h s a n x u a t p h o i b o bo in v itro \ cay chuyen phoi d u n g r o n g r a i t r o n g n h i e u p h o n g t h i n g h i e m tr e n t h i g i a i C u n g v a i s u p h a t t r i e n c u a k y t h u a t s i e u a m th u t i b a o t r u n g , k y t h u a t b a o q u a n la n h p h o i b o n g a y c a n g p h a t t r i e n v a d u o c u n g d u n g m a n g t i n h t h u a n g m a i B a o q u a n la n h p h o i b o in v itro c o v a i tr o q u a n t r o n g t r o n g v i e c b a o v e n g u o n v a t c h a t, s u p h a n c h ia c u a p h o i d u g c ta m d u n g D o d o , p h o i c o th e d u g c b a o q u a n m a k h o n g g a y b a t k y to n th u a n g d i t r u y e n n a o [ ] P h o i b o t h u d n g d u g c d o n g la n h b a n g p h u a n g p h a p d o n g la n h c h a m h o a c t h u y t i n h h o a [ ] , D o n g la n h c h a m g a y n h i e u t o n t h u a n g c h o p h o i c u a d o n g v a t c o v u d o s u h in h t h a n h t i n h t h e d a n o i b a o v a d o c t o c u a c h a t b a o v e la n h T h u y t i n h h o a la m o t q u y tr in h v a t l y m a t h e o d o d u n g d ic h c h u y e n s a n g d a n g o n d jn h n h u th u y tin h b a n g lie u d i tr u y e n tr o n g h e th o n g c h a n n u o i g ia su e tr u y e n th o n g e a c h l a m la n h n h a n h c h o n g n h u n g k h o n g c o s u h in h th a n h in v itro t i n h t h e d a [ ] Q u a tr in h t h u y t i n h h o a d u g c th ir c h i e n t r o n g d u q c sir d u n g c h o c a y c h u y e n p h o i t r e n t h e g i a i c o n g u o n d u n g d i c h t h u y t in h h o a c o c h ir a n o n g d o c a o c h a t b a o v e [ ] , K h o a n g ,5 % t r o n g t o n g s o 0 p h o i b o in v iv o la n h v d i m o t t o e d o d o n g la n h n h a n h n h a m n g a n c h a n sir d o n g la n h d u g c sir d u n g c h o c a y c h u y e n p h o i [ ] , T u y h i n h t h a n h t i n h t h e d a P h u a n g p h a p d o n g la n h t h u y t i n h h o a g o c tir p h o i d o n g la n h v a k h o a n g 0 p h o i b o n h i e n , t y l e c o c h ir a s a u c a y c h u y e n p h o i b o in v itro dong la e a c h t i e p c a n t o i iru v i c o t h e tr a n h d u g c t o n t h u a n g la n h la n h - g i a i d o n g v a n t h a p h a n s o v a i p h o i b o k h o n g d o n g v a s u h i n h t h a n h t in h t h e d a T h a i g i a n t u l u c k h u n u a c n o i la n h h o a c p h o i b o in v iv o d o n g la n h - g ia i d o n g T h a n h c o n g cua q u a tr in h b a o q u a n la n h p h o i b o in v itro p h u t h u o c v a o mot s o v e u t o , t r o n g d o c h a t l u o n g p h o i t n r a c k h i d o n g la n h va p h u a n g p h a p d o n g la n h c o v a i tr o q u a n t r o n g , a n h h u a n g (te n k h a n a n g s o n g v a p h a t t r i e n t i e p t h e o c u a p h o i s a u d o n g lanh - giai dong Vluc dich a trang thai T h u y tin h h o a c o m o t s o u u d ie m v u g t tr o i s o v d i p h u a n g p h a p d o n g la n h c h a m t r u y e n t h o n g n h u : d a n g i a n , d e t h u c h i e n , c h i p h i t h a p , k h o n g c a n c a c t h i e t b j d o n g la n h d a t t i e n , n a n g c a o h i e u q u a b a o q u a n la n h v i n g a n n g ir a d u g c s u h in h t h a n h t i n h th d d a n o i b a o P h u a n g p h a p d o n g la n h c h a m c u a b a o q u a n la n h p h o i l a d u y tr i, l u u g i u p h o i tr u y d n t h d n g t h u d n g s u d u n g d a n l e c h a t b a o v e la n h n h u h ie n c o c u a c h u n g v a c h o p h e p c h u n g p h a t tr ie n g ly c e r o l, e th y le n e g ly c o l h a y d im e th y l s u lfo x id e T r o n g k h i sau dugc T jl' £**-' b a o d e n l u c k e t t h u c q u a tr in h d o n g la n h x a y r a r a t n h a n h g ia i d o n g K h i d u g c b a o q u a n d n h ie t d o th a p , d o , p h u a n g p h a p t h u y t i n h h o a t h u d n g sir d u n g k e t h o p c a c he Email- cotihin agmail.com 64(12) 12.2022 54 Khoa hoc Nong nghiep I Chan nudi q u a n la n h p h o i Influence of the method of vitrification and developmental stages of embryos on cryopreservation efficiency of in vitro bovine embryos produced in Vietnam Khanh Van Nguyen1*, Thi Thu Huong Vu1, Thi Kim Yen Pham1 Thi Au Hoang1, Doan Lan Pham2, Quang Minh Lnn5 'K ey L aboratory o f A n im a l C ell B iotechnology, N a tio n a l In stitu te o f A n im a l S cience 2N a tio n a l In stitu te o f A n im a l Science 3M inistry o f Science a n d Technology Received 22 August 2022; accepted 30 September 2022 Abstract: The study aim ed to evaluate the influence o f different vitrification m ethods and developm ental stages o f bovine em bryos produced in vitro during cryopreservation in V ietnam In vitro bovine em bryos w ere frozen by two m ethods: (1) Vitrification by cryotop and (2) Vitrification by m icrodrop The percentage o f em bryos obtained after thaw ing o f m ethod (1) w as higher than that o f (2) (100 vs 81.01% , p0.05; respectively) In this study, the survival, cleavage, blastocyst and hatching blastocyst rates o f in vitro bovine zygotes after thaw ing were 86.83, 63.83, 21.39, and 3.91% , respectively T he authors did not observe any difference in survival and hatching b lastocyst rates after thaw ing w hile vitrification at m orula/com pact m orula and blastocyst stages (82.67 vs 88.43% , 16.54 vs 20.34% , p>0.05; respectively) T he results show ed that in vitro bovine em bryos w ere successfully frozen at the zygotes, th e m orula/com pact m orula and blastocyst stage b y vitrification Keywords: bovine em bryos produced in vitro, cryotop, m icrodrop, vitrification a g ia i d o a n p h o i n a n g c h o p h e p c a y c h u y e n p h o i n g a y sa u k h i g ia i d o n g m a k h o n g c a n p h a i n u o i p h o i in v itro s a u d o n g la n h - g i a i d o n g [ ] M o t s o n h a n g h i e n c u u n h a n t h a y , b a o q u a n la n h p h o i n a n g a g i a i d o a n serm s e t o t h e m g i a i d o a n p h o i n a n g g i a n n o [ 10 ]; v a p h o i b o d o n g la n h o g i a i d o a n - t e b a o c h i t i e p t u c p h a t tr ie n d e n g i a i d o a n p h o i d a u s a u d o n g la n h - g i a i d o n g [ ] , T u y n h i e n , n e u phoi dugc d o n g la n h - g i a i d o n g o g i a i d o a n s b m v a t i e p tuc phat trien den g i a i d o a n p h o i n a n g s a u n u o i in v itro th i viec sir dung cac phoi n a n g n a y c h o c a y c h u y e n s e g i o n g n h u su dung phoi khong dong la n h [ 1 ] Tai Viet Nam co rat it bao cao ve dong lanh phoi bo san xuat b» vitro bing phuong phap thuy tinh hoa Xuat phat tir nhu can d u e le nghien cuu dugc thuc hien nham danh gta anh huong cua phuong phap dong lanh va giai doan phat tries cua phoi bo san xuat in vitro din hieu qua bao quan lanh phoi bo m vitro Vat Bfm Tat ca cac haa chit cuu dupe mm St Louis MO, M y Sv dung cxyotop N hat Ban: pipette tao microdnop moa c Due cho qua trinh thuy tinh hoa tnm g ngfiira Nghien cuu dupe thuc hien tren phoi tinh in vitro Phirtmg phap nghien cuu Thuy tinh hoa phoi bo in vitro bang cryotop: - C h u a n b j m o i tr u d n g : M 199-Hepes co bo sung 10% s i n h (100 iu Penicillin/ml + 0,1 + M o i tr u o n g lu u p h o i b o : h u y i t th a n h th a i b e v a k h a n g m g S tr e p t o m y c in /m l) + M o i tru em g c a n b a n g : M l 9 - H e p e s c o b o ( v / v ) h u y e t th a n h th a i b e , sung 10% 10 % ( v / v ) e t h y l e n e g l y c o l 10 % (v /v ) d y m e th y ls u lfo x id e Classification number: 4.2 + M o i tr u e m g t h u y t i n h h o a : M l 9 - H e p e s co b6 sung 10 % h u y e t t h a n h t h a i b e , % e t h y l e n e glycol (v/v), 20% c h a t b a o v e la n h n h u g ly c e r o l + e t h y le n e g ly c o l; g ly c e r o l d y m e t h y ls u lfo x id e ( v /v ) , ,5 M s u c r o s e + d i m e t h y l s u l f o x i d e [ ] V i e c sir d u n g k e t h o p c a c c h a t b a o v e la n h s e n a n g c a o k h a n a n g s o n g c u a p h o i s a u d o n g la n h - g i a i d o n g T h u y t i n h h o a p h o i b a n g c r y o t o p d a d ir g e u m g d u n g t h a n h c o n g a b o veri t y l e p h o i s o n g s a u d o n g la n h - g ia i d o n g d a t > % [2 , ], V ie c lu a c h o n g ia i d o a n p h a t tr ie n c u a p h o i p h u h o p v d i q u a t r in h t h u y t i n h h o a v a n c o n n h i e u tr a n h c a i [ ], B a o 64(12) 12.2022 - T h u y tin h h o a p h o i b o b a n g c r y o to p : moi truong luu phdi can bang Phoi bo dupe d e t r o n g m o i tr u e m g c a n b a n g t r o n g vong phut sau dodm jdm s a n g m o i tr u e m g t h u y t in h h o a va dugc nap vao c r y o to p * 2 - ° C S a u d o , c r y o t o p dugc nhung ngap true d ip r ite t r o n g d u n g d j e h niter l o n g t r o n g vdng 25 giay + P h o i b o d u g c r u a la n t r o n g t r u o c k h i c h u y e n v a o m o i tr u e m g 55 j Khoa hoc Nong nghiep I Chan nudi + P h o i d ir g e b a o q u a n t r o n g d u n g d i c h n i t o l o n g c h o t d i k h i g ia i d o n g + M o i tru cm g T l : M l 9 - H e p e s c o b o s u n g % h u y e t t h a n h th a i b e v a k h a n g s i n h ( 0 iu P e n i c i l l i n / m l + ,1 m g T h u y tin h h o a p h d i b o b a n g m ic ro d ro p : S tr e p to m y c in /m l) + M o i t r u c m g T : M 19 - H e p e s c o b o s u n g ,5 M s u c r o s e , - C h u a n b i m o i tr u c m g : % h u y e t th a n h th a i b e v a k h a n g s i n h ( 0 iu P e n i c i l l i n / m l + M o i t r u b n g lir u p h o i b o : M 9 - H e p e s c o b b s u n g % + ,1 m g S t r e p t o m y c i n / m l ) h u y e t t h a n h t h a i b e v a k h a n g s i n h ( 0 iu P e n i c i l l i n / m l + ,1 - G i a i d o n g p h o i b o s a u t h u y t in h h o a b a n g m ic r o d r o p : m g S tr e p to m y c in /m l) + C r y o tu b e c o c h u a c a c g io t th u y tin h h o a c o p h o i b o + M o i tr u b n g ca n b an g: M 9 -H e p e s c o b o s u n g 10% h u y b t th a n h th a i b e ( v / v ) , 10 % e t h y l e n e g l y c o l ( v / v ) , 10 % d o n g la n h s a u k h i l a y r a k h o i d u n g d i c h n i t o l o n g n g a y la p t u c d u g c d o la i tr e n b e m a t g i a y b a c d a d u g c la m la n h - d y m e th y ls u lfo x id e (v /v ) p h u t t r e n d u n g d i c h n i t o l o n g t r u b c d o T i e p t h e o , c a c g ip t + M o i t n r b n g t h u y t in h h o a : M 9 - H e p e s c o b o s u n g 10 % h u y e t th a n h t h a i b e , % e t h y l e n e g l y c o l ( v / v ) , % t h u y t i n h h o a c o c h u a p h o i d o n g la n h d u g c c h u y e n s a n g m o i tr u c m g T b ,5 ° C t r o n g v o n g p h u t d y m e t h y ls u lfo x id e ( v /v ) v a ,5 M s u c r o s e + S a u p h u t, p h o i d u g c c h u y e n s a n g m o i tru cm g T l - T h u y t i n h h o a p h o i b o b a n g m ic r o d r o p : t r o n g v o n g p h u t d e l o a i b o h e t c a c c h a t b a o v e la n h + P h o i b o d ir g e rira la n t r o n g m o i t r u b n g lir u p h o i N u d i in v itr o p h d i b d s a u d o n g la n h - g ia i d o n g : t r u b c k h i c h u y e n v a o m o i tn r c m g c a n b a n g P h o i b o d ir g e d e - t r o n g m o i tn r c m g c a n b a n g t r o n g v o n g p h u t s a u d o c h u y e n s a n g m o i tr u c m g t h u y t i n h h o a v a d u g c h u t v a o m o t p i p e t t e , C h u a n b i m o i tn r c m g n u o i p h o i b o s a u d o n g l a n h - g i a i d o n g : m o i t r u c m g S O F c o b o s u n g ,5 % h u y e t t h a n h t h a i b e r o i t h a t h a n g t r u e t i e p l e n b e m a t g i a y b a c d a d u g c l a m la n h - N uoi in v itro p h o i b o s a u d o n g la n h - g i a i d o n g : p h o i b o - p h u t t r u b c d o tr e n d u n g d i c h n i t o l o n g d e n g a y la p t u c s a u g i a i d o n g d u g c rira la n t r o n g m o i t r u c m g n u o i S O F c o ta o th a n h c a c g io t th u y tin h h o a c o c h u a p h o i v b i lu g n g th e b o s u n g ,5 % h u y e t t h a n h t h a i b e T i e p t h e o , c h u y e n p h o i b o t i c h - p i T h b i g i a n t o lu c c h u y e n p h o i s a n g m o i tn r c m g sa n g n u o i - g ib tr o n g m o i tr u b n g n u o i S O F c o b o su n g t h u y t in h h o a c h o d e n k h i t a o g i o t t h u y t i n h h o a tr e n b e m a t h u y e t th a n h t h a i b e b d i e u k i e n ,5 ° C , % C 2, % g i a y b a c l a g ia y d o a m k h o n g k h i b a o h o a d e d a n h g ia k h a n a n g s o n g v a p h a t + C a c g io t th u y tin h h o a c o c h u a p h o i s e d u g c c h u y b n t r i e n t r b la i c u a p h o i b o s a u d o n g la n h - g i a i d o n g s a n g c a c o n g C r y o t u b e v a b a o q u a n t r o n g d u n g d i c h niter lo n g X u ly s o lie u : So lie u d u p e x u ly b a n g p h a n m em M i c r o s o f t E x c e l l v a M i n i t a b , s u s a i k h a c c o y nghT a G ia i d o n g p h d i b d s a u th u y tin h h o a b a n g c r y o to p : - va d u g c k i e m tr a b a n g yj s u d u n g h a m C H IT E S T , s u sa i k h ac c o y n gh T a v b i p < , C h u a n b i m o i tn rcm g g ia i d o n g : + M o i tn r c m g T l : M 9 - H e p e s c o b o s u n g % h u y b t t h a n h th a i b e v a k h a n g s i n h ( 0 i u P e n i c i l l i n / m l + ,1 m g S tr e p to m y c in /m l) Ket qua va ban luan Anh hifdmg cua phuongphap dong lanh den hieu qua bao quan lanh phdi bd in vitro + M o i tr u c m g T : M 9 - H e p e s c o b o s u n g ,5 M s u c r o s e , T r o n g th i n g h ie m n a y c h u n g to i s u d u n g p h u e m g p h a p % h u y e t t h a n h t h a i b e v a k h a n g s i n h ( 0 iu P e n i c i l l i n / m l d o n g la n h : ( ) T h u y t i n h h o a b a n g c r y o t o p v a ( ) T h u y tin h + ,1 m g S t r e p t o m y c i n / m l ) h o a b a n g m ic r o d r o p T a t c a c a c p h o i b o - G ia i d o n g p h o i b o s a u th u y tin h h o a b a n g c r y o to p : sa u th u tin h + C r y o to p c h u a p h o i b o d u g c la y k h o i d u n g d ic h n it o l o n g v a c h u y e n n g a y la p t u c v a o m o i tr u c m g T b ,5 ° C tr o n g v o n g p h u t + S a u p h u t p h o i d u g c c h u y e n s a n g m o i tru cm g T l G ia i d o n g p h d i b d s a u th u y tin h h o a b a n g m ic ro d ro p : C h u a n b j m o i tru cm g g ia i d o n g : KS&feNam sir d u n g c h o in v itro H i e u q u a c u a p h u e m g p h a p d o n g la n h d u g c d a n h g i a d u a t r e n c a c t i e u c h i: s o p h o i t h u d u g c , s o p h o i s o n g , s o p h o i p h a t t r i e n v a g i a n n o t r b la i , s o p h o i t h o a t m a n g s a u d o n g la n h - g i a i d o n g K e t q u a t h e h i e n b b a n g t r o n g v o n g p h u t d e l o a i b o h e t c a c c h a t b a o v e la n h - in v itr o th i n g h ie m n a y d e u b g ia i d o a n p h o i n a n g th u b n g a y th u 64(12) 12.2022 c h o t h a y , k h o n g c o s u s a i k h a c c o y n gh T a v e t y l e p h o i s o n g , p h o i g i a n n b tr b la i v a p h o i t h o a t m a n g s a u d o n g la n h - g i a i d on g doi v b i ca n h o m c r y o to p v a n h o m m ic r o d r o p (tu e m g u n g 8 , s o v b i ,8 % ; ,1 s o v b i ,4 % v a , s o v b i ,7 % , p > , ) 56 Khoa hoc Nong nghiep I Chan nuoi Bang M ri ririV ABBA A f ta t trien in vitro cua phoi bo saa■ArAAMWAmciyotop hoac microdrop in vitro Sophoi phat trifs va gi3n no' Sfi phoi i- bvbisau thoat mhng (17,79±3,87)’ Ghi chu: th "gnerr t o t e * u e : “ e- r : an lap lai, cac gia tri cung mot cct cc W mzac •n-& z iz ■ nac co y nghTa (p r-z £ ”i i r d a m b a o c h o >_• ts a m r r g - h u n g y e u t o q u a n t r o n g _ i : _ a tr in h d o n g la n h T o e - utt r ian t i e p x u c c u a p h o i i x i i c u a c h i t b a o v e la n h , q u a d o t c u t rod: g ia m th ie u c a c te n A u n i x t r o n g q u a tr in h d o n g la n h T o e d o d o n g b a h S a c s e c u p p h o t tr a n h d u p e n h fl'n g t o n th u o m g lar.h te b a o v u g t q u a a - - _ : : c h i t b a o v e la n h g iu p £ an e uek h i p c each nhanh c h o e p m a x in r a o >e d o n g la n h cr b e n a g o a L f i n la n h d o i v d i c a c g x x I r e n (-5 d en -1 °C ) m o t a n n g o a i tc b a o v a s e n a a n c h a n td n t h u e m g - f t r a x h r i c o t r o n g m a n g te baovabokhunc ~aax c d p d o n g la n h p h u t h u o c cm v a o v a t chura r r i_ d o fte s n [ ] K h i lu e m g t h e t k f c t e t t h i e u s e la m t a r c : ; m i c r o d r o p , lu p r c c _ - £ : - M a c dir k h o n g c o s u s a i k h a c c o y n g h l a v e t y l e p h o i s o n g , p h o i g i a n n d t r d la i v a p h o i t h o a t m a n g s a u d o n g la n h - g ia i d o n g , tu y n h ie n s o lu p n g p h o i n a n g b o in v itro th u d u p e s a u d o n g la n h - g i a i d o n g c u a n h o m c r y o t o p c a o h o n ifp rh d u r a m a u d u p e g i a m p < , ) T y le t h u d u p e 0 % p h o i s a u d o n g la n h - g i a i - - - - - c :a : d o n g p h o i [ ] d o n g d o i v d i p h u o n g p h a p d o n g la n h b a n g c r y o t o p d a d u p e u n fa b o a b a n g c r y o t o p v a m o t s o t a c g i a b a o c a o T h e o L O L e m e v a M A N D o d e bk % : - a ch u a m au d on g la n h c h i d a o d o n g t n a g k h a a a g - |i L V i e c sir d u n g m o t c arr gian nd va phat trien trd lai sau c o y n gh T a s o v d i m i c r o d r o p ( t u o n g u n g 0 s o v d i ,0 % , l u p n g n h o t h e t i c h d o a g # d i d a n t m h b o a s e la m t a n g t o e d o d o n g la n h n h o d vitro ± a n h p h a n d u n g d ic h -r D oi vd i ca p h u o a g p iy p A Hinh Phoi nang bo in dong lanh - giai dong - - - t- ~ r bi nut v d m ang ( ) [ ] , A D R o s a v a c s ( ) [ ] , t y le p h o i b o v itro in t h u d u p e s a u d o n g la n h - g i a i d o n g b a n g c r y o t o p d a t 0 % V a n K h a n h N g u y e n v a c s ( ) [ ] c u n g n h a n th a y v i e c m a t m o t s o l u p n g p h o i s a u d o n g la n h - g i a i d o n g b a n g p h u o n g p h a p m i c r o d r o p V i e c m a t p h o i s a u d o n g la n h - g ia i s a n g , t o n t h u o n g m a n e sm fc d a k k b i p h o t d u g c t i i p x u c tr u e d o n g b a n g p h u o n g p h a p m i c r o d r o p c o t h e la d o p h o i b i b a m t i e p v d i d u n g d ie h n : v s a u d o n g la n h - "- - - i i : s i o g c a o t y le p h o i s o n g ' - d in h v a o p ip e t t e t h u y t in h t r o n g q u a tr in h t h u y tin h h o a D o c u n g c h i n h la n h u p c d i e m c u a p h u o n g p h a p t h u y t in h B e n a n g c a o h i^ a q a a g a a a h th u s tm h h o a p h o i b o , tr o n g n g h ie n c u u n a y c fe o a e am m dune h on h o p c h it bao v e la n h e t h y l e n e g l y v o l a a d B H i q r l n l f a d d e D a y la n h u n g c h a t b a o v e la n h t h i m a d a g 06 k b d i h r p n g p h a n tir t h i p h o a b a n g m i c r o d r o p s o v d i c r y o t o p [ ] , C h in h v i v a y , t h e o q u a n d ie m c u a c h u n g to i n e n s u d u n g p h u o n g p h a p th u y t in h h o a b a n g c r y o t o p c h o q u a tr in h b a o q u a n la n h p h o i c u a n h u n g d o n g v a t n u o i c o g i a tri b a o \ e th o n g q u a v ie c A n h h i tt i n g c iia g i a i d o a n p h a t tr ie n e u a p h o i b o d e n h i e u q u a b a o q u a n la n h p h o i b o h in h t h a n h li e n k e t h y d r o g e n \o>i p h a c n r n u d e v a n g a n c h a n B e d a n h g i a a n h h u d n g c u a g i a i d o a n p h a t tr ie n p h o i b o s u h in h t h a n h t i n h t h e d a a p i b a o a o a g q u a tr in h d o n g la n h d e n h i e u q u a b a o q u a n la n h p h o i b o , t r o n g t h i n g h i e m n a y C a c c h a t b a o v e Lanh i l i a adag 06 k h a n a n g t h a m m a n g , h o a t d o n g tr o n g n o i b a o d i a o t d d [ ] N g o a i k h i stir d o n g k i t h o p c a c c h i t b a o v e la n h v d i c h u n g t o i sir d u n g p h u o n g p h a p t h u y t i n h h o a b a n g c r y o t o p n h a u t r o n g q u a tr in h t h u y m * b o a s e g r a m b d t n h u n g a n h c h o q u a tr in h d o n g la n h p h o i b o d g i a i d o a n : ( ) H o p tir h u d n g b a t lp i c u a c h u n g d o i v d i t e b a o t r o n g q u a tr in h t h u y ( g i d s a u th u t i n h t in h h o a [ ] P h o i n a n g H i e u q u a b a o q u a n la n h p h o i d e d u g c d a n h g i a IS S e NSH6 Viet Nam64