UAM_Biblioteca | Unified search engine | Scientific Production Portal | UAM Research Data Repository
Universidad Autónoma de Madrid
Biblos-e Archivo
We are onFacebookCanal BiblosYouTubeTwitterPinterestWhatsappInstagram
    • español
    • English
  • English 
    • español
    • English
  • Log in
JavaScript is disabled for your browser. Some features of this site may not work without it.

Search Biblos-e Archivo

Advanced Search

Browse

All of Biblos-e ArchivoCommunities & CollectionsBy Issue DateAuthorsTitlesSubjectsFacultiesThis CollectionBy Issue DateAuthorsTitlesSubjectsFaculties

My Account

Log inRegister

Statistics

View Usage Statistics

Help

Information about Biblos-e ArchivoI want to submit my workI want to submit my doctoral thesisFrequently Asked QuestionsCopyrightsFinancial Agencies and OA policy

UAM_Biblioteca

View Item 
  •   Biblos-e Archivo
  • 2 - Trabajos de estudiantes (tesis doctorales, TFMs, TFGs, etc.)
  • Trabajos de estudiantes (tesis doctorales, TFMs, TFGs, etc.)
  • View Item
  •   Biblos-e Archivo
  • 2 - Trabajos de estudiantes (tesis doctorales, TFMs, TFGs, etc.)
  • Trabajos de estudiantes (tesis doctorales, TFMs, TFGs, etc.)
  • View Item

Simultaneous Fluorescence and Atomic Force Microscopy to study Mechanically-Induced Bacterial Death in Real Time

Author
Del Valle García, Adrián
Advisor
Flors Ong, Cristina
Entity
UAM. Departamento de Física de la Materia Condensada; Instituto Madrileño de Estudios Avanzados en Nanociencia (IMDEA-Nanociencia)
Date
2020-09-18
Subjects
Microscopía de fluorescencia; Microscopía de fuerza atómica; Física
URI
http://hdl.handle.net/10486/693724
Note
Tesis doctoral inédita leída en la Universidad Autónoma de Madrid, Facultad de Ciencias, Departamento de Física de la Materia Condensada. Fecha de lectura: 18-09-2020

Licencia de Creative Commons
Esta obra está bajo una licencia de Creative Commons Reconocimiento-NoComercial-SinObraDerivada 4.0 Internacional.

Abstract

In the last decades, advanced imaging techniques have improved our ability to analyze biological systems at the micro and nanoscale, and in real time. Microscopy techniques have their own strengths and limitations, so their combination has the potential to provide a more comprehensive understanding of biological processes. This thesis is focused on the development and application of simultaneous fluorescence and atomic force microscopy (AFM) to study mechanically-induced bacterial death. The results reported here provide a quantitative understanding of the mechanical interactions between the AFM tip and bacteria, in the context of emerging mechano-bactericidal nanomaterials. This manuscript is divided into six chapters and one appendix. Chapter 1 provides an overview of the bacterial world and the strategies used over the years to combat the increasing bacterial contamination of surfaces, emphasizing the recent strategy based on mechanical damage. It also describes the microscopy techniques used, highlighting the strengths and weaknesses of each one, and discussing why correlative microscopy is more suitable to study this kind of processes. Chapter 2 describes the general materials and methods applied in this thesis and the software used to analyze experimental data. Chapter 3 provides the groundwork to develop a methodology to successfully combine AFM nanoindentation and fluorescence microscopy simultaneously using fluorescent polymer beads, focusing on the challenges that may arise when simultaneous measurements are performed. In Chapter 4, the methodology was adapted to image bacteria in physiological conditions, and optimal protocols to perform reproducible experiments on living bacteria were found. This optimized methodology in combination with a fluorescent cell membrane integrity marker was successfully applied to quantify the forces needed to rupture the bacterial cell wall. Moreover, a correlation between the forces exerted on bacteria and the kinetics of the fluorescence response is found. Chapter 4 is complemented by Appendix A, which provides the mechanical characterization of the bacterial wall below the rupture point, in order to give a more complete overview of the mechanical properties of the bacterial surface. Chapter 5 explores a different method to assess bacterial viability upon nanoindentation by monitoring the oscillation of the Min system, which reflects bacterial physiology. This method reveals that forces below the breakage point of the cell wall produce a fatigue effect, and provides a quantitative framework to understand low force collisions between bacteria and nanomaterials. These experiments also emphasize the limitation of integrity markers to provide a comprehensive view of bacterial response. The aim of Chapter 6 is to provide coherence and perspective to the main results of the thesis, as well as an outlook on how advanced microscopy methods and future experiments may impact the study of interactions between bacteria and nanotopographical features in the context of mechano-bactericidal nanomaterials
Show full item record

Files in this item

Thumbnail
Name
del_valle_garcia_adrian.pdf
Size
4.159Mb
Format
PDF
Description
Texto de la Tesis Doctoral

Refworks Export

Delicious Save this on Delicious

Google™ Scholar:Del Valle García, Adrián

This item appears in the following Collection(s)

  • Trabajos de estudiantes (tesis doctorales, TFMs, TFGs, etc.) [19144]

Related items

Showing items related by title, author, creator and subject.

  • Fluorescence microscopy for the in situ study of the Iberian pyrite belt subsurface geomicrobiology 

    Escudero Parada, Cristina
    2018-05-11
  • Appraisal of a Leishmania major strain stably expressingm cherry fluorescent protein for both in vitro and in vivo studies of potential drugs and vaccine against Cutaneous Leishmaniasis 

    Calvo Alvarez, Estefanía; Guerrero, Nestor Adrian; Alvarez Velilla, Raquel; Fernández Prada, Christopher; Requena Rolania, José MaríaAutoridad UAM; Punzón, Carmen; Llamas, Miguel Angel; Arévalo, Rosa M.; Rivas, Luis; Fresno, Manuel; Pérez Pertejo, Yolanda; Balaña Fouce, Rafael; Reguera, Rosa M.
    2012
  • The simultaneous low state spectral energy distribution of 1ES 2344+514 from radio to very high energies 

    Aleksić, Jelena; Antonelli, Lucio Angelo; Antoranz, Pedro; Asensio, María Carmen; Backes, Michael; Barres De Almeida, Ulisses; Barrio, Juan Abel; Bednarek, Włodek; Berger, Karsten; Bernardini, Elisa; Biland, Adrian; Blanch, Oscar; Boller, Andrea; Bonnefoy, S.; Bonnoli, Giacomo; Borla Tridon, D.; Carmona, Emiliano; Carosi, Alessandro; Carreto Fidalgo, D.; Colin, Pierre; Contreras, José Luis; Cortina, Juan; Cossio, L.; Covino, Stefano; Da Vela, P.; Dazzi, Francesco; De Angelis, Annalisa; De Caneva, Gessica; De Lotto, Barbara; Delgado, C.; Doert, Marlene; Domínguez, Alberto; Dominis Prester, D.; Doro, Michele; Eisenacher, Dorit; Ferenc, Daniel; Fruck, C.; García López, Ramón J.; Garczarczyk, Markus; Garrido Terrats, D.; Gaug, Markus; Giavitto, G.; Godinović, N.; González Muñoz, A.; Gozzini, Silvia R.; Hadamek, A.; Hadasch, Daniela; Herrero, Artemio; Hose, Jürgen; Hrupec, Dario; Jankowski, F.; Kadenius, V.; Klepser, Stefan; Knoetig, M. L.; Krähenbühl, Thomas; Krause, Julian; Kushida, Junko; Lelas, Damir; Leonardo, Elvira; Lewandowska, N.; Lombardi, Saverio; López, María Luisa Osete; López-Coto, Rubén; López-Oramas, A.; Lorenz, Eckárt; Makariev, M.; Mallot, K.; Mankuzhiyil, Nijil; Mannheim, Karl; Maraschi, Laura; Marcote, B.; Maríotti, Mosé; Martínez, Mario Ivan; Masbou, J.; Mazin, Daniel; Meucci, Mario; Miranda, José Miguel; Moldón, Javier; Moralejo, Abelardo; Munar-Adrover, P.; Nakajima, D.; Nieto, Daniel; Nilsson, Kari; Nowak, Nina; Orito, Reiko; Paiano, S.; Palatiello, M.; Paneque, David; Paoletti, Riccardo; Paredes, Josep Maria; Partini, S.; Persic, Massimo; Pilia, Maura; Prada, Francisco; Prada Moroni, Pier Giorgio Prada; Prandini, Elisa; Puljak, Ivica; Reichardt, Ignasi; Reinthal, Riho; Rhode, Wolfgang; Ribó, Marc; Rügamer, Stefan; Saggion, Antonio; Saito, Koji; Saito, Takayuki; Satalecka, Konstancja; Scalzotto, Villi; Scapin, Valeria; Shore, Steven N.; Sitarek, Julian; Spanier, Felix; Spiro, S.; Stamatescu, Victor; Stamerra, Antonio; Steinke, Burkhard; Storz, J.; Sun, S.; Surić, Tihomir; Takami, Hideki; Tavecchio, Fabrizio; Terzić, Tomislav; Tescaro, Diego; Teshima, Masahiro; Tibolla, Omar; Torres, Diego F.; Toyama, T.; Treves, Aldo; Uellenbeck, Malwina; Vogler, Patrick; Wagner, Robert Marcus; Weitzel, Quirin; Zandanel, Fabio; Zanin, Roberta; Longo, Francesco; Lucarelli, Fabrizio; Pittori, Carlotta; Vercellone, Stefano; Bastieri, Denis; Sbarra, Ca; Angelakis, Emmanouil; Fuhrmann, Lars; Nestoras, Ioannis; Krichbaum, Thomas P.; Sievers, Albrecht W.; Zensus., J. Anton; Antoniuk, Kirill A.; Berduygin, A.; Cook, K.; Kadler, Matthias; Kovalev, Yu A.; Kovalev, Yuri Yu; Krimm, Hans A.; Lister, Matthew L.; Max-Moerbeck, Walter; Pasanen, Mikko; Pushkarev, Alexander B.; Readhead, Anthony C. S.; Richards, Joseph L.; Sainio, J.; Sokolovsky, Kirill V.; Tornikoski, Merja; Tueller, Jack; Weidinger, Matthias; Wilms, J.; Bock, Rudolf K.; Bretz, Thomas; Colombo, Eduardo; Dorner, Daniela; Elsaesser, Dominik; Fonseca, Maria Victoria; Font, L; La Barbera, Antonio; Lindfors, Elina; Lozano, I.; Maneva, Galina; Mirzoyan, Razmick G.; Niedźwiecki, Andrzej; Rico, Javier; Salvati, Marco; Schultz, C.; Schweizer, Thomas; Sillanpää, Aimo; Snidaric, I.; Sobczynska, Dorota; Takalo, Leo; Temnikov, Petar; The MAGIC Collaboration; Baumgartner, Wayne; AGILE team; Fermi-LAT; F-GAMMA; CarinI, M.; Gehrels, Neil; Krauss, Felicia; Lähteenmäki, Anne; Shakhovskoy, D. N.
    2013-08-09
All the documents from Biblos-e Archivo are protected by copyrights. Some rights reserved.
Universidad Autónoma de Madrid. Biblioteca
Contact Us | Send Feedback
We are onFacebookCanal BiblosYouTubeTwitterPinterestWhatsappInstagram
 

 

All the documents from Biblos-e Archivo are protected by copyrights. Some rights reserved.
Universidad Autónoma de Madrid. Biblioteca
Contact Us | Send Feedback
We are onFacebookCanal BiblosYouTubeTwitterPinterestWhatsappInstagram