Table Of Content

AALLOONNEE IINN TTHHEE DDAARRKK Time-resolved fluorescence spectroscopy on single biomolecules Victor I. Claessen Alone in the dark Time-resolved(cid:282)uorescencespectroscopy onsinglebiomolecules VictorI.Claessen Aloneinthedark: Time-resolved(cid:282)uorescencespectroscopyonsinglebiomolecules VictorI.Claessen (cid:265)esisRadboudUniversityNijmegen-Illustrated Withreferences-WithsummaryinDutch ISBN:978-90-9026687-9 NUR-code:927 (cid:265)isthesiswasprintedongraphene-freepaper ©2012,VictorI.Claessen CarrildelZorroPublishing URL:www.aloneinthedark.nl Cover:Amodelofthecowpeachloroticmo(cid:296)leviruscontainingahorseradishperoxidase enzymemolecule,whichhasgenerated(cid:282)uorescentrhodamine6Gproductmoleculesthat haveaccumulatedontheinsideoftheviruscapsid(Chapter5). Alone in the dark Time-resolved(cid:282)uorescencespectroscopyonsinglebiomolecules Proefschri(cid:283) terverkrijgingvandegraadvandoctor aandeRadboudUniversiteitNijmegen opgezagvanderectormagni(cid:279)cusprof.mr.S.C.J.J.Kortmann, volgensbesluitvanhetcollegevandecanen inhetopenbaarteverdedigen opdonderdag21juni2012 om10.30uurprecies door VictorIljaClaessen geborenop29september1981 teAmsterdam Promotoren: Prof.dr.R.J.M.Nolte Prof.dr.A.E.Rowan Copromotor: Dr.H.Engelkamp Manuscriptcommissie: Prof.dr.S.S.Wijmenga Prof.dr.J.Ho(cid:281)ens(KULeuven) Dr.K.Blank Paranimfen: Drs.A.M.vanBuul Ir.O.Lugten Alone in the dark Time-resolved(cid:282)uorescencespectroscopyonsinglebiomolecules Doctoral(cid:265)esis toobtainthedegreeofdoctor fromRadboudUniversiteitNijmegen ontheauthorityoftherectormagni(cid:279)cusprof.dr.S.C.J.J.Kortmann, accordingtothedecisionofthecouncilofdeans tobedefendedinpublic on(cid:265)ursday,June21th2012 at10.30hours by VictorIljaClaessen bornonSeptember29th,1981 inAmsterdam Supervisors: Prof.dr.R.J.M.Nolte Prof.dr.A.E.Rowan Co-supervisor: Dr.H.Engelkamp Doctoral(cid:265)esisCommi(cid:296)ee: Prof.dr.S.S.Wijmenga Prof.dr.J.Ho(cid:281)ens(KULeuven) Dr.K.Blank Paranymphs: Drs.A.M.vanBuul Ir.O.Lugten Levéritablevoyagededécouverteneconsistepasàchercherdenouveauxpaysages, maisàavoirdenouveauxyeux. -MarcelProust Contents Contents viii 1 Anintroductiontosingle-moleculeenzymology 1 1.1 Enzymesastinybiologicaltools . . . . . . . . . . . . . . . . . . . . . . 1 1.2 Whatdowealreadyknowaboutthewayenzymeswork? . . . . . . . . . 1 1.3 Whynowindividually? . . . . . . . . . . . . . . . . . . . . . . . . . . . 3 1.4 Whathasbeendonebefore? . . . . . . . . . . . . . . . . . . . . . . . . 4 1.5 Aimandoutlineofthisthesis . . . . . . . . . . . . . . . . . . . . . . . . 6 1.6 Referencesandnotes . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6 2 Techniquesforopticalsinglemoleculeexperiments 7 2.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7 2.2 Microscopyforsingle-molecule(cid:282)uorescencedetection . . . . . . . . . . 9 2.3 Dataanalysistechniquesforsinglemoleculeexperiments . . . . . . . . . 12 2.4 (cid:265)e(cid:282)uctuatingenzymemodel . . . . . . . . . . . . . . . . . . . . . . . 20 2.5 Strategiesforsingle-enzymeexperiments . . . . . . . . . . . . . . . . . 22 2.6 Surfaceimmobilisation . . . . . . . . . . . . . . . . . . . . . . . . . . . 23 2.6.1 Non-speci(cid:279)cadsorptionoftheenzyme . . . . . . . . . . . . . . 23 2.6.2 Site-directedenzymeadsorptionviaa“proteinfoot” . . . . . . . 24 2.6.3 Site-speci(cid:279)candcovalentenzymeimmobilisation . . . . . . . . 25 2.6.4 Enzymeimmobilisationviareceptor-ligandinteractions . . . . . 27 2.6.5 Immobilisationofthesubstrate . . . . . . . . . . . . . . . . . . 28 2.7 Spatialcon(cid:279)nement . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29 2.7.1 Encapsulationindroplets . . . . . . . . . . . . . . . . . . . . . 29 2.7.2 Femtoliterarray . . . . . . . . . . . . . . . . . . . . . . . . . . 30 2.8 Flowassayinacapillary . . . . . . . . . . . . . . . . . . . . . . . . . . 32 2.9 Referencesandnotes . . . . . . . . . . . . . . . . . . . . . . . . . . . . 34 3 Insolution:studiesonfreelydiﬀusingenzymemolecules 39 3.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 39 3.1.1 (cid:265)econtainer . . . . . . . . . . . . . . . . . . . . . . . . . . . 41 viii

Description:

MICROSCOPY FOR SINGLE-MOLECULE FLUORESCENCE DETECTION. 9 experiments—is Total Internal Re ection Fluorescence microscopy Scienti c writing could bene t from the introduction of evidentiality—as it exists in.

Alone in the dark: Time-resolved fluorescence spectroscopy on single biomolecules PDF

160 Pages·2012·5.91 MB·English

by Victor I. Claessen

Checking for file health...

Save to my drive

Quick download

Download

Upgrade Premium

Most books are stored in the elastic cloud where traffic is expensive. For this reason, we have a limit on daily download.

Download Alone in the dark: Time-resolved fluorescence spectroscopy on single biomolecules PDF Free - Full Version

by Victor I. Claessen| 2012| 160 pages| 5.91| English

Download Alone in the dark: Time-resolved fluorescence spectroscopy on single biomolecules by Victor I. Claessen in PDF format completely FREE. No registration required, no payment needed. Get instant access to this valuable resource on PDFdrive.to!

Free Download PDF

About Alone in the dark: Time-resolved fluorescence spectroscopy on single biomolecules

Detailed Information

Author:	Victor I. Claessen
Publication Year:	2012
Pages:	160
Language:	English
File Size:	5.91
Format:	PDF
Price:	FREE

Download Free PDF

Safe & Secure Download - No registration required

Why Choose PDFdrive for Your Free Alone in the dark: Time-resolved fluorescence spectroscopy on single biomolecules Download?

100% Free: No hidden fees or subscriptions required for one book every day.
No Registration: Immediate access is available without creating accounts for one book every day.
Safe and Secure: Clean downloads without malware or viruses
Multiple Formats: PDF, MOBI, Mpub,... optimized for all devices
Educational Resource: Supporting knowledge sharing and learning

Frequently Asked Questions

Is it really free to download Alone in the dark: Time-resolved fluorescence spectroscopy on single biomolecules PDF?

Yes, on https://PDFdrive.to you can download Alone in the dark: Time-resolved fluorescence spectroscopy on single biomolecules by Victor I. Claessen completely free. We don't require any payment, subscription, or registration to access this PDF file. For 3 books every day.

How can I read Alone in the dark: Time-resolved fluorescence spectroscopy on single biomolecules on my mobile device?

After downloading Alone in the dark: Time-resolved fluorescence spectroscopy on single biomolecules PDF, you can open it with any PDF reader app on your phone or tablet. We recommend using Adobe Acrobat Reader, Apple Books, or Google Play Books for the best reading experience.

Is this the full version of Alone in the dark: Time-resolved fluorescence spectroscopy on single biomolecules?

Yes, this is the complete PDF version of Alone in the dark: Time-resolved fluorescence spectroscopy on single biomolecules by Victor I. Claessen. You will be able to read the entire content as in the printed version without missing any pages.

Is it legal to download Alone in the dark: Time-resolved fluorescence spectroscopy on single biomolecules PDF for free?

https://PDFdrive.to provides links to free educational resources available online. We do not store any files on our servers. Please be aware of copyright laws in your country before downloading.

The materials shared are intended for research, educational, and personal use in accordance with fair use principles.