Data reproducibility in fluorescence image analysis.

Texte intégral

(1)Data reproducibility in fluorescence image analysis. Catherine Souchier, Christine Brisson, Bernadette Batteux, Michel Robert-Nicoud, Paul-André Bryon. To cite this version: Catherine Souchier, Christine Brisson, Bernadette Batteux, Michel Robert-Nicoud, Paul-André Bryon. Data reproducibility in fluorescence image analysis.. Methods in Cell Science, Springer Verlag, 2003, 25 (3-4), pp.195-200. �inserm-00333348�. HAL Id: inserm-00333348 https://www.hal.inserm.fr/inserm-00333348 Submitted on 23 Oct 2008. HAL is a multi-disciplinary open access archive for the deposit and dissemination of scientific research documents, whether they are published or not. The documents may come from teaching and research institutions in France or abroad, or from public or private research centers.. L’archive ouverte pluridisciplinaire HAL, est destinée au dépôt et à la diffusion de documents scientifiques de niveau recherche, publiés ou non, émanant des établissements d’enseignement et de recherche français ou étrangers, des laboratoires publics ou privés..

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(332) \ $+(* ¨T 'X Y. [+WYlX

(333) \.

(334) \Yl \d$ Yr0. ]\"dZYZ r§ \Ylsya YZ

(335) sKX\ k^d * \[s Xdr g ) \Y$d`¨. (

(336) *k d. § ¬Yy. +(o a. §Xd. *\ k \[k. \YHZ(, g d$ \[ / $ \[a. X\[k. $ k.

(337) \"d . $od g [dU d$ ( YZ

(338) \"dU,7xB(* n,XdZYZ $ YlX -k,

(339) \ aX)(*( § YZ

(340) \ $ Xd$

(341) \ / X)[\Y$d r( $. $ § d $. $ X g kmX\ km Xdr $ kfd$ $X(Yy

(342) d¨. 7. x. d \ § Xd< Xdr $ k Y YZ

(343) d< \ Y§ k ]Zl $ \Yk X0.d`¨"7 §X)\[X(]0"d$ bd § Xd. lZ, $§ k. X Yl .

(344) X/ mXYb $ d d$

(345) \/^dg , $X( \[ G YZ $X( r *(Yl . dg GY$d. X\ k.

(346) \Yl / $XGYl

(347) ,\ Yy

(348) dX\ kªYZ[ ¥ X\ y( $ d$ \ ¥

(349) \Yl \.d$ Yl * d $ drYlX\[k X $k k X\[k n $ dEdE k

(350) \YZ[ d$X S\ YE¨ 7 } $ dr(Y$d § g dy"§X $ k

(351) \ X+(* [dU>Yl ¦~Z cYZ § * $"d $ dBX\ kª

(352) \ X+(* ¡ªZ cYZ dg,( YZ

(353) \.dS.'x9(* nX " ¨L7. k *X)\ X( bd§d$ >\ ]\¥YZ[ (*X[dZYB,(¬"§\¨ N

(354) DdrYla(

(355) "d$ ~k XGYlX $ +YlX ]\[ k

(356) \k \ k \Y,\3YZ k X0. ¥YZ ,\[k, Yl

(357) .\d X" bd$ Yl

(358) \"¨9p , $ , $a Xd. Gn GYl k aYZ ]\Yl \"d$ YZ0O

(359) \ $ Xd$ k X $k

(360) \/ YlYZ X§[\Y§ k[0, d,\YZ[ $od g [d X\ k9YlSYZ[ Tx9(* nX} \ )\YZ $XGYZ

(361) \.d ,"YZ 9d$ ( Yl

(362) \.d¨. bXG+(

(363) }9Y ¦ X l[no¨[ $ . . bXG+(

(364) }¡HX $n¨ $ . E DD*# .

(365)

(366) . . XGYlX § $ +YlX

(367) \[ k X) Yl $ X $XYZ

(368) ,\ X)\[k X\X(]0"d$ bd c|l\.d ¥+, Xk.d XGYk

(369) yl $ \YYZ

(370) d}k $

(371) \/ X $

(372) k~. YZW $ §§.\YZd¨,N drYla Y<d$ "(

(373) k + \ Yl k YZ[XGY YZ[. $ dZ"(Y$d § $ m+YlX

(374) \ kf"\[k K $ / $[d. \YZ l ( ,Hr$d$ GY. X\ k. +[ V

(375) /YZ\[ dd¨W [ d$ "X $X GYl Ed § $ Sl -n k X\ k d$ GYw ]\>YZ[ 9d$. YX $ ¨ \ k a§. $. X $ "(\[[Y'Y ª $ X YZ ¥d$XYb $XGYZ

(376) \ ( (

(377) aX\k YZ Xdr $ \Y$d" $. YN \BYZ T $X\W/" " s YZ

(378) \9YZ ""¦ ¦B/ $XG0d$X(* ¨. . }\YZ § (* ac g yk +(* drYlX\[k.X gk, W kOk XGYlX $ +YlX

(379) \[ k aT , \ \9YZ[ S\[ G YZ $X(.Z (¬Yl d$ GY$d , YZ T

(380) \Yl / gXGYZ

(381) \9YZ

(382) d $ }§.k

(383) r * k"¨ $a $ dr(Y$d" $ + YbYl TZ cYZ[ / $ \ X\ k¥ $ kd$ V * dgawY X\ Z, TYZ[ +( \ ¨N [. ,X)( dU[+WYlX

(384) \ k Y § \YZd ¢ . Z LYZ . +( B. \ \Y § y >d$ /\ y *X\Yl(0 (

(385) X Yl . dgX (* ¤$a>X\[k YZ bd [+dg XGYZ

(386) \ Xd \ y $ ki+ 03b rYZ[ . dg $

(387) dB,< Xdr g ) \Y$d`¨L7 (

(388) "dDdB TZ(, g d$ \[ YZ f +(¬ +, Xk"d. Xd'\ YU $ (

(389) XGYl kYZ¥ Gn $

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(391) \YlX)( bdrYlX aSYZ O . , \ \Y ,§YZ +( x^dgX) (* a. Xdr g k§X9d$ \ k9YZ

(392) }¢ e<¤ § XdL\ k +Yl k (0§X . , \ \YE¨. 7 lXGYYZ XGY r( $ d$ \ d \.d¥X0H[X)\ / Ok $

(393) \/iYl

(394) bd. $+X+(]0. YZ[ m X

(395) \. X, gk ]\W/. Yl. k $X +X. Z d$

(396) \ /. YZ . XdQdrYlX\[k X $k"¨ $a. [[ Gn $

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(398) .

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(401) *k d X $ \[k [ +WYl k,(03dE (r¨}pO $ ga \ "\[ n GYl k , X)( d ¥y(, g d$ \[

(402) \Yl \d$ Yr0 X $ §[+WYlX)

(403) \[ k aLX§\ . dZYlX\ kX $k~§X0 +, g X $ k~X\ k¥drYl d$ ( Yy -,\d. >Z(- l.W $ § d §X0O+ drYl $ k

(404) \YZ k X y k y

(405) \/ d$ , $X( d r

(406) D¨7|y\.

(407)

(408) . Xk k Yl

(409) \ aLxB(Yb/.(*XdBd$( *k dj X0~+ )Zy * * )\Yy(0 Nd$ k~$, +,X / $\ k~ y $ DYZ

(410) \ X\[k§, $X(

(411) ( dy0drYl K,\YZ y (b¨ . /.\

(412) y *X\Y y $ (

(413) XYy

(414) \.d $ O[+WYlX ]\[ ki+ Y§ \3 § Xdr $ \Y$d X\[k. YZ. (* (bd. Z(, g d$ \YH ]\Yl \"d$ YZ0. , $afYZ. §Xd. $ (

(415) XGYl

(416) \. \[[Y. d y * k d YZ l

(417) GYl(-0. |y\d . + Xkod`¨. E&[, bYZ

(418) \ X(b¨mN. rYZ . Z, 3YZ. Xdr g ) \Y$dgaL Y § (

(419) k +, "dDd$ +(* Yl¥, Z $ Y7YZ §k XGYlXa.YlX

(420) \/

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(422) \W/ YZ > X/ >X\ X(]0d$ bd dy0drYl >¤. X)\[k. YZ[. YZ[ , g `YZ

(423) X( , X)(o d. ¢ pO(

(424) G(bX. vN y+ dy¤¨3T. $ dr(Y$d. k § \"drYZ $XDYl cYZ XY"YZ }

(425) X/ }X\X(]0d$ d'dy0drYl g *k d7 $ g.k +(* }X)\[k (

(426) \ X k XGYlXW¨¥YlX\[k X $k XGYl

(427) \. d$

(428) \ /. \[ G YZ $X(¥y *(Yl Dd Xd d$ >\Q ]\. "dDd$ +(* ¨ | Y XdX(bd$~ \ r

(429) $ k+ 0~ n GYl k X\Yy

(430) Z *XYy

(431) \ "dDd$ + (

(432) YZ0. YZ bd drYbk0. Xd. $ dr(Y$d' \ y\

(433) \ / Z(, g d$ \[. E D¨T7 $ Z, $ a § X ªYl \.d$ *k YZ[. .\(* X jt x. YZ XY%YZ 9YZ r X( d YZ[ y( $ d$ )\ +, Xk.d § $ aLX Yl }d$(

(434) *k.. $ X $XGYZ

(435) \ a'\ Y. GnXGYy(0YZ dgX YZ[X)\O

(436) \idg,(¬ Yy

(437) \OX\ k"\[k >YZ X\[X)(

(438) 0"dE bd. . \ k Yl

(439) \.ddg k + 0 pO(

(440) G(*X vN $+ d`¨[p , g gao Yw d'

(441) rYlX\YY dbYlXDYl jYZ[XGY \ ( (bd}X\[k Yl bdDdr d$ Yy

(442) \"d<X $ X\ X(]0 k a g "X $XGYZ § YZ k.dT>NdrYX)(*d$ +, ¥ \YZ $ (

(443) ( k acX\ k+

(444) .(

(445) /. -X( X V *XGYl

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(447) \Yl X[\YB \ªYZ[ dgX) (

(448) ]\ /k d$

(449) /\§X\ k§ ]\BYZ } ]\Yl $ YlX`YZ

(450) \§ "YZ }r(o $ d$ \Ywk XDYlX¨ xB Yl >drYlX\[k X $k XGYl

(451) ,\[a \ X $(-0 *k \Yl

(452) X(k.XGYlX $ +YlX ]\[ k§ \ YZ dgX). d ) * )\"d. $. X\[X(]0W k. X Yl Q X\W/.

(453) \/. \[ G YZ $X( y (Yl . dg GY$dQ.

(454) \Yl / $XGYl

(455) ,\§YZ

(456) d¨ bd' bdYZ[ > X

(457) \ ,\[)(Nd$

(458) .\ ,LY d § y¨|l\ k k a Y \d YZ[ "dDd$ +

(459) ( YZ03Z ¥§ X[dZ $ ]\W/3X\ k .X V

(460) \ /Hd ] \"d "\[k &YZ ] k \ k Yl

(461) \.d$a \ § [ \ \ (]0¥Wsr+ YU X/ §X0 +, §X " g k"¨§§ $a\ G YZ gX( y

(462) (Yg . d l XGYy -,\d^d$ "(-k. +,. $

(463) Nd$(]0. ) k. d$

(464) \ /. d$ \Yy

(465) X(.

(466)

(467) X " bd$ Yl

(468) \"d¨>7 dZYbk[0 § Xd $Z V k d$

(469) \ /3YZ TX)\Yy

(470) GY ¡

(471) X/ X\[X(]0W `¨ § § $a d$

(472) (*X VZ $§X\ dO[(

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(474) (Y > YZ . \§X( *XGYl

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(476). j¨ \0 aX X)\ ¨ U *Xsy§ * $"dy0"drYl d$a GY (

(477) X $a9 VX)\0. 0W+X *k97Ylk a. k k

(478) \/Yl \9p -k k,(* d$ na §¨ >.

(479). l . ¨ ¨. "¨ ¨. ej X,X¥9a ( $vwaUo $X\ .(

(480)

(481) \ p aU.§Oc¨ UZy Y$d§ V *y * k $ ,>+

(482) \X\Y§\[ G $X(}X\[k >Nd$(* X/ $

(483) \ ,\ dy ( YlX(9>dg)(* y + Ed

(484) \ ¨'&' )(

(485) ( e}

(486) (b¨ L N¦W 7sV¦ ¨ ej $s ](

(487) (~&'a } $XX ]\3pvwa>pO $(

(488) \DUap X $ X( a T"

(489) (

(490) (* §

(491) \ ¨ GYl y§

(492) \XGYl

(493) \~,

(494) \YZ $X (

(495) ("(*X < y/X\ (

(496) (* dT

(497) .(* *XGYl k

(498) \~k XG\[, Z + *\ 0Yl& (-X[d$§ *ªdg drYZ $XDYZ

(499) \

(500) \>(Yy *k r/sZ $ d$ bdrYlX\YTp &Nsy (

(501) ( d}Nd$

(502) \ / y( $ dg \ § * $"d$ 0 * X/ X\ X(]0dD bd`¨'&0Yl. GYZ r0¨ < ¡ s "¦¨ N DdZYl a q [0âHSX )X)(3vU¨ n V * )\Y$d d$

(503) \/ § * $X y $X0 Yl \ (

(504) ,/ 0o ( * YlXYy

(505) \.d X\ k drYlX\ k X $k & gXGYZ

(506) \/ $ k g d¨ U\ k y

(507) \ (b¨ M N . .¦Ws ¨ ,(

(508) d }UaSUX\Y 7p ¨vN $Yl

(509) \ (*X+ (* ]\ / § ¬Yy y( $ d$ )\Y y+ d`¨ p GYZ k.d&' (]( e} -.(b¨ L [¡ ¦Ws ¨.

(510).

(511)

(512) ` ,\W §xBaTB rY$d a<&c * vwapO y§X\OxS acUX) Ed §a X\ t , gk \m& ¨ e}Xd$

(513) dZYZ $XGYl /

(514) d3Z ,X(] *km0Yl. GYZ r0 d$

(515) \ /

(516) X/ X\[X(]0"dE bd¨ § bdrY .[ ¨ sy¡¨ ` , ejaB7[dEd$ \ xBa !BUa GYl \ a (0[.\"dy 0 a q .(

(517) (Tv à "( (* &pOaej )\[od> ¨ q /(*XGYZ

(518) \ . ¬Yy[ )(* *X)('tX¢r£¤[X)\ \[ ( d+.0 XYZ ]\

(519) \ (*X)\X 9(

(520) . *k + (*X0 DdBX\ k ]\~YZ )\[& dS .G0Yl Gn $ dd$

(521) \ dy0drYl ë}

(522) (&c ¨b NW ."¦WsV¦¨ U ]\[Y*YlsD X rY aªvwXDY Yl Edy \ ¨ § (

(523) \Y X\ k d$ y( $ dg \Y lYl

(524) \§ X y Dd

(525) \§(]

(526) \ / )(*(bd¨.

(527) \ ¨ [ ,W syL¨ p .k (pOxBae' y [X $kY ¨x drYlX\ k X $kOZ, X(

(528) + EXGYZ -,\OX\ k d$ Xk,

(529) \ / Z $ GYZ

(530) \ }X r( $ dg \ * $"d$& ¨N&0Yl § YZ r0¨ L L NL s ¡"¨ t GY Yl \ §a"'(

(531) ( \+, y/ BU a [\W/ |gc¨ ¡"¨.|y(

(532) ("§

(533) \XGYl

(534) \~X( + $XGYl

(535) \ Z >Z(- $ dg \ ¥ $odg 0"+¨ |$\Yl y\ XGYZ -,\X(Tdg. * GYZ0Z X)\X(0Yy *X( 0Yl,(-,/ 0¨ (b¨[ (* \Y'WaoxT+drYZ $XGY'¦W "¨&U0Yl GYZ r0[a q

(536)

(537) \ a"|rYlX(-0o¨ "¨ vwX\[)s .(

(538) "dy

(539) \X ta §X/.(*X\ k q vwa ej bd$ sDYl §X Y a e< X(

(540) /XGY p apO

(541) ( (*X $kªv apOXNaU G\ / ¨x9(* nX k0 d$a'Xªdg $ d§.B\ y( $ dg \Y k[0[ d YZ XGY 0[ * )(*k n, Yl

(542) \ X( (

(543) 0 + l ]/Yla YZ"dbYlXG+(* \«r/XGYl d¨ § dZYl.[ H&0Yl. ~¨ W sV ¨

(544) U&'a,Nb $ \ >pOa,ej \ X + p a"&'XGYlX(

(545) (

(546) q a,e< g0[ \>vLx§¨pO GYZ[.k.d Z H ( ( $.(

(547)

(548) y $XGYl

(549) ,\ X\ X(

(550) 0"d$ bd + 0 y( $ d$ \Y X/ 0Yl, GYZ r0¨ &0Yl § GYZ y0¨b .¦ sy "¨ /a X(

(551) §.\Dp a UX 0 a *X( (* GY v¨ L( $ dg \YH

(552) X/ 0Yl, GYZ y0.b g. X( YlXYy }Yl X\YZ YlXGYZ > Xdr g ) )\Y$d¨ xB\ X(7&' (b( vwXGYZ[,(b¨b L [ "¦Wsy¡.¦¨ a 9(bd'v7Ba7} XG X gX)(* q |$a e< $X \ [.r< a"tX\ \

(553) \ /X9ta *dE)[ tB .(

(554) k $

(555) \ /.&U<¨|$§X/ }0Yl GYZ y

(556) } GYZ kZ X\YZ ]b0

(557) \ />YZ T $ (*XGYl , X§\Y , jt x^

(558) \ +XGYl y *X( \(* .

(559) k.d¥d$

(560) \ /3}d$[ $ *X .(

(561) b¨ pO * $"d$W¨ N ¡¡Lsy¡.[¨. ¦¨.

(562). ¡"¨. "¨.

(563). ¨ "¨. "¨. ¨.

(564). ¨ "¨. "#. %. * "!$. 7 bd § yXddr [, bYl k9+ 0 x q &

(565) Vx>ddg.

(566) XGYl

(567) \§Z, q dg X $§&'X\ 7/ $X\YE¨.

(568)

(569) . XG+(* ¨|l\.d X(] + $XYy

(570) \>+ XkZ XGYb g d. q Z $ \ . p ,(* G(*X LvN y+ d. §

(571) X GYl . n,) ¬YlXGYZ

(572) \ bdd$

(573) \. |$sl 7. |y\ >e}(. ¨-¦ . .¦W>\ >\ . |$sl 7. |y\ S g \. ¨-¦ . ¦W.¦B\ 7¦¦B\ . |$sl . |y\ S gX\/. ¨-¦ . ¦>\ 7¦W¡ >\ .

(574)

(575) . XG+(* "¨)N( $ dg \ }Z (Yl dg GY$d U§ dd$ -.\> $X\W/". n,) ¬YlXGYZ

(576) \Z (¬Yl .

(577) $. -cpO y $. § bdd$

(578) .\§Z (Yl . ej(. ecv ¡ >\ . Bp. >\ . 7v "¦B\ . } $ \. ecv >\ . Bp. ¦>\ . 7v¦¦WsV¦W¡ >\ . q k. ecv¦¦WsV¦W¡ >\ . Bp. ¦ W>\ . 7v¦W >\ . e'v)ejX\ k9vwXdEd$a9pi)§ * $

(579) cpO

(580) y $ $a7v) \/>vwXdEd.

(581)

(582) ¦. X+( }"¨pO Xdr $ § \Y7 / $ \§Z(, $ d$ \YN§ * $"d [ $ d.

(583) ! " $# %"&(') +* 7 8 '5, " 8 "<', 8 ;8 1<' "+ * 8 *9'5" D " #6: ' " & ? . ",%-.', /*0", 12', ," 3" "4'5, 6- ," %".', # ", - ! 19') * ;: ') -<'! -<') 1 , 1 / = ' / ' * ' - 8># ' ? *, - 19' "-<'5" * '" ;+8 ' # 1 @8 # *9' # #/ 68 ' : #A; : ' : * 8> ' #A :BC 1 A 19' * 8> 19'*/1 * 2- ' " ,:: ' / / 6::BE 'F ;GH,GIGJ9K,L F.

(584)

(585) ¡. XG+(* T¨)p XdZ $ § \Yw, $ kZ(, g d$ \YL§ * $"d $ d. ) " 7 " "+ .

(586) ? ; & ')%" ' ) " A '! '! " " 8 '! * " 8 ' ) # 19'! # *9' 7# ' 7 A ' "/-2'" " 8 '5" "+1/'" 8 *9'* * ' # * # *9' # 8 **9' " 8>## ' 8 8>#68 ' 1 ->6- 8 ' * ** 68 ' #6: - :># ' * 8 * * # - '5- : & ? 'F ;GH, GIGJ9K,L F . "<'! *9'! : /1'" * *' 8: ' 1 *8"# ' :-. ) , * " 8B , :+ 7#

(587) C " * * @: * 8># 1 = =8 - * 8 " #?E .

(588)

(589) . XG+(* 9¦¨pO Xdr $ § \Yw "+(- }Z(- $ dg \YN§ * $"d $ d. ! " 7 " "+ . L?

(590) *! # & ' ) 8 8 '5,%". L? "3 : ', ", : ' , 8 *2', 8 ' , * :/: ' 16-<' = '* ' -#' 7# ' #' " ,8 +'* =# 19'*6" / "<'* # = " # '5-+1 & ? ; 'F >GH,L G@GJ9K6/L F. "3 "/', 1! *<'5, * #6: ' %" - '* */1+' " 8 *6-<'* #. . ; "3 '5, "3 :9 ! " *! 8 '5, 8 B ! :+ 1 >' 1 ?# %- 8 ' = . ! : / 6: '* ,:B ! - /* * ' / " E .

(591)

(592) . XG+(* }¡"¨pO Xdr $ § \Yw x9(* nXT" §d$( YZ

(593) \"d. ) - ) 16&('! )%- "3 ".', 2', , -3 8 ', ,",%- = ' ) /1 - 1! *2' ) : & ? 'F ;GH,GIGJ9K,L F. ) : ' , ', # ', %-.', 1)%".', . )%-.', ', 2', *)%".', 68 # ', 6" ", 12'! *6". / ; ! 1 *! " 8 # -3 -.

(594)

(595) . . N

(596) / $ ¨xB" bd$ Yl

(597) \9d$

(598) \ / \[ G YZ $X(r *(Yl d. ¨§&', Z $ (*XGYZ

(599) \ + GY \ r( $ d$ )\[ i $XGYZ - dg \Yl

(600) X(K Xdr g ) \Y. d$

(601) \ /^Y§. X\ kK\ G YZ gX)( Z (¬Yl $XYy

(602) a X Yl . . k

(603) rl $ \YQ\[ G YZ $X( Z (¬Yl Dd¨ >¨ . d \.d g § XdZ $ k d$ ]\ / \ G YZ $X(Nr *(Yl 9dg GY$d § YZ~XYbYl \XGYZ -,\~b $.. Y H¢Vd

(604) \HX¤ X)\[k Z $. dZYlX\ k X gk, W k"¨. ¨. . . Yl ¨ ¢Vd )

(605) \ + ¤¨TXGYlX $ YZ \. N

(606) / $ "¨xB" bd$ Yl

(607) \9d$

(608) \ / nodZ $ YZ

(609) d. § d ] \.d § g S XdZ $ k9d$

(610) \ / nodZ $ YZ

(611) db $, Y. SY d. ¢Vd

(612) \§X¤LYZª "dw¢Vd ] \>+ ¤¨X`YlX $ YZ \ dyYlX\ k X $k k"¨.

(613)

(614) . 1.0. Filter ratio. 0.8. 0.6. 0.4. y = 0.96 x + 0.04. 0.2. 0.0 0.0. 0.2. 0.4. 0.6. 0.8. 1.0. Fluorescence ratio. 500 ND14 400. ND11.2. 300. ND8. 200. ND5.6 ND4. 100. ND2.8 0. A. B .

(615)

(616) 7. 20 t=2 15. t=4. 10. t=8 t = 12. 5. t = 24 0. A. B.

(617)