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Non-invasive manipulation of membrane potential in intact living cells

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Material Information

Title:
Non-invasive manipulation of membrane potential in intact living cells
Physical Description:
Book
Language:
English
Creator:
Dando, Robin
Publisher:
University of South Florida
Place of Publication:
Tampa, Fla
Publication Date:

Subjects

Subjects / Keywords:
Biophysics
Electrophysiology
Na
K ATPase
Electric fields
Ionic concentration
Fluorescence
Dissertations, Academic -- Applied Physics -- Doctoral -- USF   ( lcsh )
Genre:
bibliography   ( marcgt )
theses   ( marcgt )
non-fiction   ( marcgt )

Notes

Abstract:
ABSTRACT: All living cells contain the electrogenic enzyme Na/K ATPase, whose function is to pump ions against the electrochemical gradient, in order to provide potential energy which is later used for cellular processes such as action potentials, muscle contraction and facilitated transport. Using a technique developed in our lab, exploiting the molecule's voltage dependence, it is possible to increase this pump function by many folds. Optical measurement of the membrane potential of living cells was made using a potentiommetric dye, with successful manipulation of the ionic concentration and membrane potential reported. Additional supporting results are presented, along with extension of this field to the study of cardiac Myocytes, representing a progression to Mammalian cells, with advantages to future clinical research evident. Successful manipulation of membrane potential was also achieved using cells in a two dimensional tissue matrix, which more closely approximates the living system, and hence is closer to an eventual clinical application. Also, expedited recovery from electrical injury was recorded, demonstrating a possible therapeutic application of the technique.
Thesis:
Dissertation (Ph.D.)--University of South Florida, 2007.
Bibliography:
Includes bibliographical references.
System Details:
System requirements: World Wide Web browser and PDF reader.
System Details:
Mode of access: World Wide Web.
Statement of Responsibility:
by Robin Dando.
General Note:
Title from PDF of title page.
General Note:
Document formatted into pages; contains 150 pages.
General Note:
Includes vita.

Record Information

Source Institution:
University of South Florida Library
Holding Location:
University of South Florida
Rights Management:
All applicable rights reserved by the source institution and holding location.
Resource Identifier:
aleph - 001935456
oclc - 226064135
usfldc doi - E14-SFE0002171
usfldc handle - e14.2171
System ID:
SFS0026489:00001


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Full Text

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85 r"rn"rrn"r nr&nn rnrrrrnrr "rrnrrnn rnrnr "r nnrrrnnr rnrrn nnnnn nr "r nnrnr"rrr nrr nrr)n rn rn$rrn! )rrrnr rnnnrrn n"rn"rrrE nrnr)nrnn rn "nrrnnnrn n nrnrrn"rnrr rnn7r rnrnnn rrnnr7 /7nnnr nrrrr nn)nn */7rrnr "nr n nnrr /nr nn7r n rnnn rnn"n177( r!n nnnnnn "nrn177( r1rn rnE"r*77nn /77rrn rnr"n rnrnrr n%r/8n"rn" rn nnr"n2"nrn) "nr

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88 rnnnrn"nrn rnrr" rrrnrrr )n*nnn nn%&n$10 $10 $10/ $10 $100 1$ 1$ 1$ 1$ 1$ $$$$$$$$$$$$$ +#3n@ rrr r%4 r$%4 $ rrnn rr"r rrrrnrnnr rrn rnrr nn nnnrn

PAGE 77

84 rr"r nr" nnr"rr rrrnn nnrnnnr nr rnn nnrnn" nrnrn"rn nrnnn" $rnn nnnnr r r"nrrnn "n rnrnnr nnnrn nnnnn! nnr n nnnrnnrr nn nn%r/4 rrnn n"rnnn$nn57

PAGE 78

8= )n*nn n $1 $1 $1 $1/ $1 $10 1 $$$$$$$$$$ )n&nn 7* 53%4 53%4 53%4 53%4 #5nrrr$ r$0%4 % %r/4nr)rn rrnnrr nrnr rrn rrnnnn rr1rrnr n76*7@rn rrrnn r;rnnrn .nrnnnr nnr rrr )nr"nrnnr nrnn"nrn n17@$nrn$rn nr#r nnrnrr rnnr n nnr!rnr n nr"rrn

PAGE 79

83 nnnr"r n!rnr n nnrrnn nnn rnn nnrn nn"nrr rnnr"n r nnnn$n nnrnr n rr n nrnnn rn

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47 rrrnnrrnn nrnErnr"r rnnnrnrnnn rnnrn"n nrrEr>r )rrn nnn nn r nnr" rnrnn !nnnrr "rnnn ;nrnr#rn "rn )rrrrnn n! )r

PAGE 81

4 n)rnrnnrr nnr n n!nnn r!rnr nnnnrn rrrnr rrnr nnnrnn r"nrnrrn rnrrnr'(r,Hrr$n))nnnr nrn) rnn'" nrn" nrn!n n!nnrn nnnnrnr rnn"r rnr)nnnnr rnrr nnr0>
PAGE 82

4* r"nnrnr n$nr r!' "nrnnrn" nrrnrn rnrnnr nr"rr>D>r n rr"rnrn r nrnnr"rnrr nnrn$nrE *757n"rnrn "rnnrrrn nrrr rnnrnn r "nnnr nrnrrn nnnr

PAGE 83

4/ 0r!r2nrr""nrnn nnnr n rnnrn! nrrn$nn 1U>rnr$ rn,n$n& 331A(nrrn3=8A#n335A 2nn345-nnrn nnrnnr"r"r nn nnn nnn *nnn r&r'nr nn/4U>5@>H*rn r/787n15r" >nnr n577:#nrnr rnn rnn"rnr (002n rn35V"rrn nr "rnnnnr2 !nrnnn rnr nr"r rn)rnn rn#:0>r r)nrnr"n nnrn "rrnn 77Vrrr! Enrr"r rr"r rrnr nr#?

PAGE 84

41 r(002?=n>7(00214> 5>n>*#2H1 *(*H158< >nn?n(5@ >nnA 0)rn?n(V##:0 #KA >r,9#0#5@%'2n@rrn(41n*5U>

PAGE 85

45 'r?>rrnnn "r7nnrr" n"nnrnrnn >rnnr In.0&4*%rnrn$ nnrrnn r(nrnnHK=nr %" nnn$nr9nn rn$"r/7 n)nnn" n )rnnn!r rnrr87 57( r nrn r7nnn nn=7.n$n$ nr!n r57177( rn@"r7 rnn n4*rr"rn nrr "nr)rrn$n nrrr

PAGE 86

48 'r;r(nrn)rnrr nrnrrn r"nn nr rnr$nr)n nr)n n"nrrnrnnr 0>rn)r rnnnr n &nr "rrrn nrn"rr rnn rnn r"nnnrrn$ n"n "nnnn"n n"r&n")r r")nn nnnn "nn"rnnnrnnrrn nrnrn" r nrr"rnr r "rrnnnnr rn%r1

PAGE 87

44 *n:rr rr nn$nrn nnnn rnnrr nrnn nErnrr "r)r rrn"rn n"rr n" rrnr nnn rnnrrnnr "nrrn rn"rnnnrnn rr rr nnrnr r"nn$nn$n )n"nr "nrn"rn n$r 89

PAGE 88

4= rrn"rnn rrBrC nn$nrn$"rn n$)rnnrn r"nr n"rn rrnrnrn rrnn rnnn "rnrnrr!rr n$rn"nn rnr%r1*nnn n$)rnnr rnnnrnn"nrn n ) n.*/nnrnn$ $$ $$$ $$ $$$ $$ $$$ $$ $$$ $$$$$$$$$$$$$ nnn :2* 5;**<) *n.02 rr

PAGE 89

43 &4nnrrnrn$ nrnr rnnr nnr nr rnr rnn$n$r )rnnrn$ r7*7nr rnnnn !"nrn nnr nn"nnn n nnrnnrnr)n rnn$r nrrnnn r"n nrnnn$ nr)r rn$n"rr )n r nn)n nnnr nn"n rnr%r1/

PAGE 90

=7 ) n$ /$ $ 0$ $$ $ $ $$$$$$$$$$$$$/$$$$ nn "$ "$ $ $ $ $ *#n
PAGE 91

= nnn n$ /$ $ 0$ $$ $ $ $$$$$$$$$$$$$/$$$$ nn 3%4 **n>r r$ Drnnnr nn nn "$r r"rn%r #r"rrnn nnr"nrn nrrn nnr rrnnrrrnnr n$rr"nn

PAGE 92

=* nnrnnrn n) nrrnnn nnnn nrnn nrrn nnnnn nrr nnrnnrrn rnrnn r"n" nnnrnn "rr"rn. nnn rrnnrnrrnnrn rnnn nn rn)rn nnr nrrnnnrn r!n177( n n n"n57( nnnrnn rnrn nr!nnnr nrnnr! rnrnrn $nrn r"rnnn nnrnrn nrr"nr n n&nr) nr"nn nrn r nnnr!r )rnnr rrnrnr %r15?

PAGE 93

=/ )(. n$ /$ $ 0$ $$ $ $ $$$$$$$$$$$$$/$$$$0$$ nn 0$ $ 0$ $$ 0$ $ *1n
PAGE 94

=1 n(. n$ /$ $ 0$ $$ $ $ $$$$$$$$$$$$$/$$$$0$$ nn '<2)1 *3n>r%rrr r%rr0$r rrrnr"%r18r nnrn nnr nn nr nnrn nnrn >r"rn rrnnnrn nnr"$nr ,-n

PAGE 95

=5 n,-nrr n nnr rrrn$nr nnnnn%r14 rnnrn n? 0(. 1$ /$ $ 0$ $$ $ $ $$$$$$$$$$$$$$ nn 3%4 '<2)1 *5n
PAGE 96

=8 nn r"rrnr rnnrnrnrr n$nnn nnnnn $nrn rrnnnnnrr r"nnn n$nrnn nrr" n nr nnrr nrrrr n%r1= )n'nn$ $ $ $ $ $$ $ $ $ $$$$$$$$$$$ nn *7n>%4 r2 r %4r r%

PAGE 97

=4 2")rnrn rnnnr "rnrr nrn rnnnrnr nn nr"n )rnr%r13 nnn)n'nn$ $ $ $ $ $$ $ $ $ $$$$$$$$$$$ nn *8n>rrr%4 r nr%r13n"rrnnr "nnr"n "nrnnr r rnnr

PAGE 98

== rnnr"nrn rnrrr rn"rnrnr nr/7nn n*7@nnrrrrn $nrrnrr )r

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=3 +rrnr"nrrn nrrr nrnnrrn" rnnn rr rrn rnnr) rnrnn rnnrrHr rnnnnr" rn nnnn r nn nr"n nnnnn nnnn"nrn rnn"nn nn2!nn" rnrn nrnrn" n"n$nr nnnnr nrn

PAGE 100

37 &n')nnnr nnnnn rn$r*nn nrnr rn"rnn rHnnr"nrr nrrrrrnr r!r" !nnrnnrnr "rrrnn" n)nr "rrrnn &'(r,'nnnrrnrn nrnnnnn "rnnr" "rr rn"rn rrnn nrrn)rnnrr nrn rnrr nrr nnr)nn n"r"n D nnrrn n$rnnr rnnrnrrnnrnr "n"r>D>

PAGE 101

3 nn)nn n"nrrn rnn)rn nrrrn "n!nrnr$ rn$!n rnnn$rrn nn rnrrnrnr,( rn*77/An$n *777A>n'nnn*77A'$nn33 =-%r5nn nn"nr"n 2rnnr" "nnrnrr" rn nnnrn"n"n r)rn

PAGE 102

3* 1n!rrrrrr % r $ $r nrn"61602nnnnr$ rrrn"n nr rnnnrn nrr&rn nrn nnr) r%r5*

PAGE 103

3/ 1n>rr$r #n nr"" nn" rrnnnnrnBr$ nnC rnrn #:0n%r 5/nrnnrnn

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31 1#nrrrrr r %rrr nrnnnn rrnr""r rnn Inrrnnnrn nnr"rn "rnrnn r"nnr n$nr rnrnrnrnr" nrnnn rrr"!nnr$n nn %r51

PAGE 105

35 1*nrrrr nnnrr"r nnnn rrn"rnr"rnnr nnr*7 rnn rnnrrnnr r"nr rnr rr n/@nnrr E%r nnrnn r"nrn rrn" nrnr

PAGE 106

38 &0r!r0r#r21577n rn nr"nn$rnn2rn 9nrn *77n/77rnrnrr"n nnr 2nnnr!$r rnrnn n'r2n,'2-)n 35@)5@>H* nnrrnnrnnrr" rrrnr rnnr nr rrnrn nn rnn"5n rn!nnn !rr'2 rnH21577 nrn$rrr %rr nn57rrr n!nnn rnn $nr)nn rnrnrr nn2nr nrrnrnr$nr n$rK r"rrn

PAGE 107

34 r(002?=n>7(00214> 5>n>*#2H1 *(*H158< >nn?n(5@ >nnA 0)rn?n(V##:0 #KA >r,9#0#5@%'2n@rrn(41n*5U>

PAGE 108

3= & 'r2nrn "rnrrrrn nrnrnrrn n9<0.*77 "nnnrrrr "nnr "nnrrnnnr ,"nnnErr nnn nrrnnr rnnrn"n"nn "n"r nnrn$ "rnnr$nrr"n nrn" "rnnrn$5 7rn$nr nnnn r)nn nnnn /r1nnrnrn nn %r55

PAGE 109

33 11n!rrrr %% $"rn rnrnn rnrr rr nnr r"rnn rrn$ nn nr/r1rnn$ nn rnnrnn /1n rn$rrnn rrn

PAGE 110

77 "n nnrn"nrnr nn"nrnn rrn *77.""nnr "nr nr/7.nnn

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7 &'r;r(nr"nn!nn nnnrn )rnr nrnr nnnn nrnn E"rn rnr$nrnr nnnn nrnrn rnn rnnnr nrrrrnn rn "nnrn%n rnn!nnrrr"r n nnnrn nnrn )rnn rnnn nrrr"r "r/nn" rrr$n nD nr%r58

PAGE 112

7* 13n>r nr)rnr "nrnnnr nrnrnr "rr"r0rn nrnnnrrrr En ))%n 5n*n **) :%44 n )

PAGE 113

7/ &&4#nnnrnrn$rr )rnr nrrnn$rn nnrrn"!n &Dnr$n rr)rn "nnnn"nnr "nnrn nn!nrn"rnn nnrn"rn rnrrrr !nnr rrr n"nn n!nnn"nr nr"nrnn nn$n#Hn"nn rrnr rnrrnrn rnn"n nrrrnrr rn$nn nrrrrr n ;rnnr"n n%r54

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71 nr n$1 $1 $10 $10 1$ 1 $$$$$$$$$$$$$$ +15n!rrr$ n"rn"rr"r nnr"n rnnnrnrn nnn nnnnrn rnnnrn )rnnr n rrrnnnn r nn &)nrnn rnn "rrnn r)n r nnn r$n$nrn$

PAGE 115

75 nr!r177( /7( nnr!n rrnr%r5= ) n(. n$1 $1 $10 $10 1$ 1 $$$$$$$$$$$$$$ +17n!rrr0$r nnn"rn"rnnr "nn r"rnrnn nnnn nrnnr" rnrn nrrnnn rnnn nrnrn"nr nnrr r nnn"r r"rrnnn

PAGE 116

78 rrrrnn"n nnrrn nrnrnr% r53 ) n0(. n$1 $1 $10 $10 1$ 1 $$$$$$$$$$$$$$ +3%4 '<2)%n* 18n!rrr0$r rnrnr nn"rn nnrnrn$nrn rnrr nr nnnn nnrn n$nnnnrn nnrr rrn"n nr nnrn r"r

PAGE 117

74 nnrn$nrnr nn nrnr"rnnrn r$Ennrrnn rn&nn nnrnn r rr!nn r nr!n )r177( r nrn nnnr! n/7( n nrn nr!n rnn rnrr"r' nnrrrnr nnn )n-nn r$10 $10 1$ 1 $$$$$$$$$$$$$$ +19n!rrrrr$

PAGE 118

7= rnnr/@"%r57 rrn rrnr nnnnr &n n!n)nnn nrrnr rnn n)"n "rn"nn*77. rrr rn)nn 1n!rrrrr

PAGE 119

73 n)*77.n"nnr" nn nrnnnr nnnnrn nn("rrnrnr nn$nnr 57n n/757rr rn/5$n !nnn rnr8n7rnnr *77.nnr 2!n)n) nnrn rnnr/7.n n)n"nn r"nn*7. rrrn rn nrrnrnr nnrnn nnn rrr$n nrnnnr n"nrrr nnrnnn nrnrr nn nn"rn rnnrrn nnrrn rr "nnnnr) nnnrnnr )rnr nrnr rrrrn rr rr"rr nrr nnrrnrn rnn nJ0rrnnrr nnnr nr"nnr nnrrnr

PAGE 120

7 rrnnnr rnrO nrnrnrnrn nrrnrnr n r"nnn"n nr)nrnr nrnrrn nnnrn Er)$nnnr)r n)n r"nrnnrn nnnr nnEr nrnrrnr rnnrnn nrnn n"n rr "nr nnrn *77.nnrnnr nnn rn%r5*

PAGE 121

1nD?r rrn)r nn n"r*r/ nrn nrn nrrn nrnn rr)r nnrnn rn)rn nrnrn nn rnn) rnrr nnrr"rnr n nn"rnr rnrnr n )rn nrr)rnnrn rrnnrn

PAGE 122

* nnrnrr"n rnr nn$r nrnnrnrr n"rrnn/7 n)r(r5n "r nnnn nnn$ rnnrrn rn$r nrrr nn nnnrnr rnnrrnn r%r5/

PAGE 123

/ ,r,0(. 0 n n0$ 0 0 0 0 $$ $ $ $ $ $ $$$$$$$$$$nn n =n!nn =n!nn3%4 =n!nn'<2) 1#n!rrr rrr )rn57@ rnr nn 5n& rnnr nrrrnr n r"nrn rrrnnn "rrnr )rnrrnr r"r rIn.0&nnrrn nnrrn nrnnn nrrn$n rnnrrrrn nn"rrr

PAGE 124

1 n"n)r r"rn rrnrnnn"nn rnnnr nrnrn nrnn r"nnnnr rnnrnr nrr"rnnnr nnr"n nn57.rn rrnnr nrnrn "nrrrrr nrnrnnn rn nrn )n'2! nnrn nrrrnrn nrn/4U> n5@>H*rrnnnrn0" rnnnrrn rnn nErnnn rrn("r rnr$r"n nrnr" rnr nn%r1

PAGE 125

5 nr n -nn0$ 0 $$ $ $ $ $$$$$$$$$$$$$/$$$$0$$$$$ +1*n
PAGE 126

8 &+rr2nrrnrn nr rrr"nr nnr n nrrn3@r rr nnrnr )nnrrr nnnrnn$ r"nrnr/7 nnnrn*7@rn nr5 n)r nrr nrnr"r $rn n)rnrn nrrr$n nnr nrr n"nnrn(r rrnn nrn"rrnrn n nnr nn! rrrrn rrnrrnrnnnn

PAGE 127

4 +rrnnr"rr rrn n"rn*7@rnrn nrnrn7@ nnrn !nrn rnnnnrrnr nnrnr"r rnnn+'(nrrn nn rnn"rnnn "rnn n&n"rnr rr$n r nn!n nn rnnrnnn "%r8r rrnrrnnn rrnr"nr nrrnrnn

PAGE 128

= ,nnnn,r ,$ $ $ n0 >! >)n*)n*n!* 3n! r%r n%r8n"nn rnrrn" nrn$nn ;rrrn "rnrnnr rn&r rn$rnrn r)nrnnr" $rn n rnnnrrn r)nrnE

PAGE 129

3 nnrnnnr$rr rnr nrnrn rnrnrn nnnnrnnn"r n&)n rnnn nr)rn rrrn rn)n nrrnnr rnnrn nrnrnnrnr rnnEn n$rn )r !nnrrrrr nrrr) nnnr nnr) nnnrrrn nnn n"rrrnn rrrr nnrnr nr nE"rnnn )nrnrn nr nn nr rnEr n"nnrn nrnnrnn nrnn nr nrnnn rn" BCr"n rnnnn n" rrn nrnnnrr rr"rr r"n

PAGE 130

*7 n! n"rrn nnEr rnrrn 6"n"rn nrnrrr nrrn!n nnn"r4n rn("r rnrnnr n)rnrnrnr" n"rnnn Hrnrnnrrn" nnrnnn&n" r"rrrrnn n$nrnn" )nn"rr""nr nnnn nn$rnErr "rHr!nn r"rr$n"nrn rnr rnn "rnn0)rr n$nn nr)rnrnn nnnrrn rrn!n rnnr)n rnnn"r rrnrnr $nrrr rr! rnnr"rnnn nrn rnrnrnr r"rrn rrr nnnn nnrr! rnErD2n rnrnrr*nr nnrnEr r6nnEr:"rr 6nnErnr

PAGE 131

* r"rnrnn nr$)rnr nnrn rnnrnn r&rnrn nn)r"rn rnnr! rr rnr rrrn r"nnr r"rrrnErnn rrrrnn nnnnn"n r $rnn$rnrrn r"r$nnr rrnrn" rrrnrnn rrnrnrnrn rnrnnrn "rnnn r

PAGE 132

** +'.>r)n"nnn nrn)r nnrnrnr nrnnr)n nn"rnr n""rn nrnrrn )nrn "rn" )r nr"rn r rn+nn nrrnnn nrrrnr nnr n"nnrn nnnr rn! rnrnnnn Inrr!rrnn nnrnn !r"nrn "rnrn nr"nnn61602 nrn rnnnrrrn$ nn nnrrnnrr nnrr "rnrnr nnrnnn n"nrrn9>n rnn r>nn rrr

PAGE 133

*/ "nnrrnnr ""nn nrnrnnnrn nrrn )rnnr nnr nnrnn

PAGE 134

*1 !rnrn" r$ >N9n2r n#nn #n (rnr #rn:n n%r E;r ;r ,*774-/3?46*8 >N9n0rn"nn nrn>rn N9n2r n#nn n(rnr #rnn#nnn>nrn> =;>E ,rr"

PAGE 135

*5 ?r,4, ,:nnn%4@ 'nnn"rnrA:" 8+/864*46458%AB nrnrnrnrr n6n +F> :: 3=8?// %#40#r#!#r# O>O9n&r(. +> 8/D2=1?1/161/= %#40 rnnn'rnr > 88.*48?34%40 nnnr"n .%r $ :: .3,-=7*,1

PAGE 136

*8 r1!#?#r@.4B >n#rn rr2n":n (nn2+r 88/# =?1547615=4 #'rr#? 0nrnrrnn ;; 8/8 **?//6/** @ 2r n#n>nrrr6#n 6rnrr> #rn ::@#@@ A nrrr6$6.r" n rnrrr$nr ;r% :: 58?336*7/ @#66' 2r nnnrn E ;r; ::+ @#66'#A.# 0rn#rnr 2r n#n0r%' n+rn,r r"-#@#?#4 r".nn"nn rrn$nr'n +rn 88 88?4776473

PAGE 137

*4 C'r' 2'n9rnn6'A ::: .1=A =8/6=47 # >nnrnnn <> 88/ 35?/64 r%4' 9rn9rn"" >0 69%r:"r nrn,096 %:-'+ ::# =7?*7=6*7*= 0@!#;(0#4r>4. .nn6 .%r 8// 57?**56*1 #r1#@0#@B!#?r >? #rn nnr"n>r rn9r >n9A'+rnA 8// .5/A4=56431 ;(0#"r 2n6nrr6"nrn n n"rnr ED.%r 8/8 31?565/4 ;!B;( 2r2+ 8+-# 3*?*46*/5

PAGE 138

*= ;( r#'nn #Irn,=r r%r r :n" rOr$ 8/ //61= ;rE?r>? %rr#rnn?#r rr nn#>'A./7A 8/8 A3/6*= Ar'%#4,(0A#;##;; nrnr n'">nrn#<#6 rnAn :rr>nr9r#nrnr n3=8*3?578651 Ar#$#4="')$ =0@ H rn62"n>nn rr/6rr6 rnrn%n>rrn n:n2r$ ::# /1?75 Ar%?AF(%. Rnn"9r#rn>rrn n>n0)nr" E.%r 8&# 45776511 $#!#@r#' ()nrnr)nrr '"rnrr"nrn "nrn)n) #rrn 88 ,-1?*/6*/8

PAGE 139

*3 $r>r>%B#'''#4B.; 0"nr rnrnnrn r'6#n '6*+> :::# *43?>=5*6>=53 ?!#%AB rrrr6"nn" r n6 =;%rE 8/+ /?/736/* ??#A?#1'?#B0 r(B I6n nr:H2rnrnrrn $nn )rA2n A ::# .1/A/7P1 ?r>?# >n#rr%r9 A#>'A 88 ./=356*73 r0'#'r#?1C#B#'r ;4#6r @G#rr#A(""' '6*%n#rn %n0rrnr>n:nH) 9r"n6> 9nA9n"2#>$nA#nrn> nr'A :: 31671 rr#'%#;B4#r0' ;r<;B rn nrrErnrnnr 6"nn r'#26=711=>nr"nnr:nr 88+ /?3/67

PAGE 140

/7 "r#;(0 WnXnWXnrr6 "n rnn"rnr ED.%r 8/8 31?5/36585 nr 26nrrrrrrn r"nrr nrn !rA nn6n nrn rn)rnn,nE!; 8// /1?856=* !r4?Br(! I$n2nnn" rnrnr(n0rr#rn' 'n 8&-# *5?=6*=!r4?#A(<(%$' "nnrn rn$nn rnrnrnn +'> 8*14?85/768517 4=r%'r' 2nrIn nn"#r :nrAA#'>A :: A.1,=-A//=3P//33

PAGE 141

/ 4r>4.#;(0#0@! .nn E '; 8855?756** 4r>4.#1?%#?rB#'>)@ n" rr6"nrnnK rr )rnWX E'; 88 *?46=4 4(#@B# ;#1#"rrr B!#'r4 !9A >n6 n"nn"n rn nnrnnrrnrnnnr rnn2 ::: 34A/*8P//4rr##%#0 %r!nn rnn n$n E<.%r 88 31,-?411641= 4rr4#)%#.. r4''' #rnn)n"rnn nrrn"n /6rrn?%n.7%+)*@ :: n@>))% @$4r#%#1#4B Drnnr"n n>> 88 173?84168=5

PAGE 142

/* '# A#r#C 2nnnn6":Ln$ nrrn)rnr E';% 8/ 41?36 *7'!B#@r%'#@A#Ar,,B. 2n nnrrnnr n n"'rA 8-# ./A//56///7 ''BE"rr;? nrr rn > -& ?4*76/34* 'rB nnnr nnrrrn r"''n 8&*/,*-?/31617 '"!# B 9"n"rnrr 6 ?nrn6nnn n .rr;%rr G';% :: =5?/=46175 '#;;#.B',. nn >nnrnrnrn nn r"rn''n 8-& 177?8*688

PAGE 143

// #B#r#C 0".n6nn nnrrrn E'; 8/: 55?//617 rC##A >nn'rnrnn r rn ..>; :: /,*-?*6= @r#%' 9nrrnn:"' 8-8 .=?=146=8= @r%' Hnrrnn+#r' 8-+ *4,1-?/4P //1@<#B#%#40# r""rn$r ?rn > 88: *14?153 @. >nr' 88+ #r

PAGE 144

/1 %

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/5 %F%n0 n(%"rn

PAGE 146

/8 rrnn nn"r"nrn rnrnn)n nn $

PAGE 147

/4 %Fn?1 @ !r In.0&rrnr r$nr nrr6rnnn rrnn r!rrnr !"nn nr*rrnrrnn nrn" r! nrnrn nnnrrn%nnnrn nrn rrnnnrnrn nnrnn rrnr" nnrn nnn"nnnr rrnrrn "nrnnn,rnrn n$nr-n nnnn"nn rrrn rrnrrrn n"nrn n")nn n nrrrnr "rrn 2!nr rn 57( nnrn rrnn nrnr!n r!rn rr!"nr nr! rrnn

PAGE 148

/= "rrrnr$n rnrn nrnrn$n rnn"rn !rr"rnn n

PAGE 149

/3 #n;0

PAGE 150

17 *n

PAGE 151

1 1n;0

PAGE 152

1* 3n;0

PAGE 153

1/ %F(!rr(rn)nrn" )n Dnn$nr 7*7" rnrnrn"n rnnr nn$nHrn nnr!r" nnnrrn rn nn"nnrrn( "rr"nr nrnrrn$n!nn" nnrnn rnrn n)rn nn

PAGE 154

11 %F0n!rr)rnrrnr rr>* nn$rrnnnn rrnnrn nnrrr rrn r"nr rrnr nrrr nrnrnrnnnn rnrrr$ nrrn nn nnr"r/7nrn" n nnrnrrnn r<%rr"rnrn nrnrn n"n rn>* rnHrrn nnrnnrnr nr6)n )n"r rnrn rrnrrrnn )nnrrr nnnrnn rnn rnrnnnrn nrn$r nnrnnrn <%nrn rn>*r rn)rnnr

PAGE 155

15 rnDrn"r rrrn rnnnr) nr rnrn$r nr "rnn

PAGE 156

18 %F n rrr.r"nrrnrn nrrnn"rn r"rn & *)& n -nn$10 $10 1$ 1$ 1$ 1$ $$$$$$$$$$$ +5n@ rr$ %$

PAGE 157

14 %F.n *nnn rnnr"n !rrrnr rnrr" nrnn r 7n
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Dando, Robin.
0 245
Non-invasive manipulation of membrane potential in intact living cells
h [electronic resource] /
by Robin Dando.
260
[Tampa, Fla] :
b University of South Florida,
2007.
3 520
ABSTRACT: All living cells contain the electrogenic enzyme Na/K ATPase, whose function is to pump ions against the electrochemical gradient, in order to provide potential energy which is later used for cellular processes such as action potentials, muscle contraction and facilitated transport. Using a technique developed in our lab, exploiting the molecule's voltage dependence, it is possible to increase this pump function by many folds. Optical measurement of the membrane potential of living cells was made using a potentiommetric dye, with successful manipulation of the ionic concentration and membrane potential reported. Additional supporting results are presented, along with extension of this field to the study of cardiac Myocytes, representing a progression to Mammalian cells, with advantages to future clinical research evident. Successful manipulation of membrane potential was also achieved using cells in a two dimensional tissue matrix, which more closely approximates the living system, and hence is closer to an eventual clinical application. Also, expedited recovery from electrical injury was recorded, demonstrating a possible therapeutic application of the technique.
502
Dissertation (Ph.D.)--University of South Florida, 2007.
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Includes bibliographical references.
516
Text (Electronic dissertation) in PDF format.
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System requirements: World Wide Web browser and PDF reader.
Mode of access: World Wide Web.
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Title from PDF of title page.
Document formatted into pages; contains 150 pages.
Includes vita.
590
Adviser: Wei Chen, Ph.D.
653
Biophysics.
Electrophysiology.
Na.
K ATPase.
Electric fields.
Ionic concentration.
Fluorescence.
690
Dissertations, Academic
z USF
x Applied Physics
Doctoral.
773
t USF Electronic Theses and Dissertations.
4 856
u http://digital.lib.usf.edu/?e14.2171