title: High-fidelity optical reporting of neuronal electrical activity with an ultrafast fluorescent voltage sensor.
abstract: Accurate optical reporting of electrical activity in genetically defined neuronal populations is a long-standing goal in neuroscience. We developed Accelerated Sensor of Action Potentials 1 (ASAP1), a voltage sensor design in which a circularly permuted green fluorescent protein is inserted in an extracellular loop of a voltage-sensing domain, rendering fluorescence responsive to membrane potential. ASAP1 demonstrated on and off kinetics of ∼ 2 ms, reliably detected single action potentials and subthreshold potential changes, and tracked trains of action potential waveforms up to 200 Hz in single trials. With a favorable combination of brightness, dynamic range and speed, ASAP1 enables continuous monitoring of membrane potential in neurons at kilohertz frame rates using standard epifluorescence microscopy.
authors.affiliation: 1] Department of Bioengineering, Stanford University, Stanford, California, USA. [2] Department of Pediatrics, Stanford University, Stanford, California, USA.
authors.forename: François
authors.initials: F
authors.lastname: St-Pierre
authors.affiliation: 1] James H. Clark Center, Stanford University, Stanford, California, USA. [2] CNC Program, Stanford University, Palo Alto, California, USA.
authors.forename: Jesse D
authors.initials: JD
authors.lastname: Marshall
authors.affiliation: 1] Department of Bioengineering, Stanford University, Stanford, California, USA. [2] Department of Pediatrics, Stanford University, Stanford, California, USA.
authors.forename: Ying
authors.initials: Y
authors.lastname: Yang
authors.affiliation: 1] James H. Clark Center, Stanford University, Stanford, California, USA. [2] CNC Program, Stanford University, Palo Alto, California, USA.
authors.forename: Yiyang
authors.initials: Y
authors.lastname: Gong
authors.affiliation: 1] James H. Clark Center, Stanford University, Stanford, California, USA. [2] CNC Program, Stanford University, Palo Alto, California, USA. [3] Howard Hughes Medical Institute, Stanford University, Stanford, California, USA.
authors.forename: Mark J
authors.initials: MJ
authors.lastname: Schnitzer
authors.affiliation: 1] Department of Bioengineering, Stanford University, Stanford, California, USA. [2] Department of Pediatrics, Stanford University, Stanford, California, USA.
authors.forename: Michael Z
authors.initials: MZ
authors.lastname: Lin
mesh_terms: D000200:Action Potentials / Q000502:physiology*; D000818:Animals; D002478:Cells, Cultured; D002645:Chickens; D002938:Ciona intestinalis; D005260:Female; D005456:Fluorescent Dyes; D057809:HEK293 Cells; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D009474:Neurons / Q000737:chemistry / Q000502:physiology*; D011247:Pregnancy; D051381:Rats; D017207:Rats, Sprague-Dawley; D013997:Time Factors; D056969:Voltage-Sensitive Dye Imaging / Q000379:methods*; D014982:Xenopus laevis; D015027:Zebrafish
chemical_list: D005456:Fluorescent Dyes
references.citation: Nat Neurosci. 2012 Dec;15(12):1742-51
references.pmid: 23103964
references.citation: J Physiol. 2013 Sep 15;591(18):4427-37
references.pmid: 23836686
references.citation: Neuroscientist. 2009 Dec;15(6):625-34
references.pmid: 19321459
references.citation: J Neurophysiol. 2012 Oct;108(8):2323-37
references.pmid: 22815406
references.citation: Front Mol Neurosci. 2013 Mar 04;6:2
references.pmid: 23459413
references.citation: Curr Protoc Mol Biol. 2010 Oct;Chapter 14:Unit14.20
references.pmid: 20890901
references.citation: Biochemistry. 2009 Feb 10;48(5):929-40
references.pmid: 19140681
references.citation: Proc Natl Acad Sci U S A. 2000 Jan 18;97(2):901-6
references.pmid: 10639177
references.citation: Cell. 2013 Aug 15;154(4):904-13
references.pmid: 23932121
references.citation: PLoS One. 2012;7(9):e43454
references.pmid: 22970127
references.citation: Nat Biotechnol. 2005 Jan;23(1):102-7
references.pmid: 15580262
references.citation: Chem Biol. 2009 Nov 25;16(11):1169-79
references.pmid: 19942140
references.citation: J Neurosci Methods. 2008 Jun 30;171(2):296-308
references.pmid: 18471891
references.citation: Proc Natl Acad Sci U S A. 2013 Apr 9;110(15):5939-44
references.pmid: 23530193
references.citation: Nat Biotechnol. 2006 Jan;24(1):79-88
references.pmid: 16369541
references.citation: PLoS One. 2008 Jun 25;3(6):e2514
references.pmid: 18575613
references.citation: J Neurosci Methods. 2007 Mar 30;161(1):32-8
references.pmid: 17126911
references.citation: Neuron. 1997 Oct;19(4):735-41
references.pmid: 9354320
references.citation: Nat Methods. 2011 Nov 27;9(1):90-5
references.pmid: 22120467
references.citation: Biophys J. 2002 Jan;82(1 Pt 1):509-16
references.pmid: 11751337
references.citation: J Biol Chem. 2008 Mar 21;283(12):7983-93
references.pmid: 18160398
references.citation: Nat Methods. 2009 Dec;6(12):875-81
references.pmid: 19898485
references.citation: Chem Biol. 2009 Dec 24;16(12):1268-77
references.pmid: 20064437
references.citation: Proc Natl Acad Sci U S A. 2008 Jun 17;105(24):8428-33
references.pmid: 18550841
references.citation: J Neurosci. 2012 Oct 3;32(40):13819-40
references.pmid: 23035093
references.citation: Nature. 2013 Jul 18;499(7458):295-300
references.pmid: 23868258
references.citation: Nat Methods. 2007 Sep;4(9):741-6
references.pmid: 17721542
references.citation: Science. 2012 Apr 13;336(6078):229-33
references.pmid: 22499946
references.citation: Nat Rev Neurosci. 2000 Dec;1(3):181-90
references.pmid: 11257906
references.citation: Hum Mol Genet. 2013 May 15;22(10):2105-18
references.pmid: 23406872
references.citation: Curr Protoc Neurosci. 2003 Aug;Chapter 6:Unit 6.17
references.pmid: 18428582
references.citation: Cereb Cortex. 2011 Jul;21(7):1645-58
references.pmid: 21084454
references.citation: Neuron. 2011 Mar 10;69(5):885-92
references.pmid: 21382549
references.citation: J Neurophysiol. 2000 Nov;84(5):2398-408
references.pmid: 11067982
references.citation: Nat Methods. 2012 Oct;9(10):1005-12
references.pmid: 22961245
references.citation: Nat Rev Neurosci. 2012 Oct;13(10):687-700
references.pmid: 22931891
references.citation: Neuron. 2012 Sep 6;75(5):779-85
references.pmid: 22958819
references.citation: PLoS One. 2013 May 22;8(5):e63067
references.pmid: 23717400
references.citation: PLoS One. 2013 Jun 19;8(6):e66959
references.pmid: 23840563
references.citation: Nat Protoc. 2006;1(2):695-700
references.pmid: 17406298
references.citation: Front Neuroeng. 2009 Oct 13;2:14
references.pmid: 19862342
references.citation: Nat Struct Mol Biol. 2014 Mar;21(3):244-52
references.pmid: 24487958
references.citation: Nat Neurosci. 2012 Mar 25;15(5):769-75
references.pmid: 22446878
references.citation: PLoS One. 2007 May 09;2(5):e440
references.pmid: 17487283