Principle and basis of AFM technique

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Table of contents

1. Scientific context
1.1 Biological membranes
1.1.2. Eukaryotic and prokaryotic cell membrane structures
1.1.3. Phospholipids
1.1.3.1 Phosphoglycerides
1.1.3.2 Physical states of membrane phospholipids
1.1.4. Lipopolysaccharides
1.2. From a single bacterium to bacterial biofilms
1.3. Antimicrobial peptides
1.3.1. Polymyxins
1.3.2. Bovine Catestatin
1.4 Application of lipid membrane and bacterial biofilm models on peptide studies
1.4.1 Langmuir monolayers
1.4.2 Atomic force microscopy
1.4.3 Attenuated total reflection Fourier transform infrared spectroscopy (ATR-FTIR)
1.5 Aim of this work
2. Materials and methods
2.1. Preparation of phospholipid and lipopolysaccharide bilayers
2.2. Bacterial culture and formation of bacterial biofilms
2.2.1. E. coli bacterial strain
2.2.2 Determination of minimal inhibitory concentration
2.2.3. Bacterial culture
2.2.3. Bacterial growth in suspension
2.2.1. Formation of bacterial biofilms
2.2.4. Fluorescence microscopy and BacLight ™ staining
2.3. Langmuir method
2.3.1. Compression isotherms
2.3.2. Brewster angle microscopy (BAM)
2.3.3. Preparation of Langmuir monolayers and compression isotherms
2.4. Atomic force microscopy (AFM)
2.4.1. Principle and basis of AFM technique
2.4.2 Force spectroscopy
2.4.3. AFM for phospholipid and LPS bilayer experiments
2.4.4. AFM imaging and force spectra of biofilms
2.5. Attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR)
2.5.1. Principles of ATR-FTIR
2.5.2. ATR-FTIR on phospholipid bilayers
2.5.3. ATR-FTIR on bacterial biofilms subjected to AMPs
3. Antimicrobial peptide action on model membranes
3.1 Phospholipid monolayer interaction with a cyclic antimicrobial peptide
3.2 Phospholipid bilayer interaction with antimicrobial peptides
3.2.1 Morphological and spectral characteristics of phospholipid bilayers
3.2.2 Phospholipid bilayer interaction with a cyclic antimicrobial peptide
3.2.3 Phospholipid bilayer interaction with a linear antimicrobial peptide
3.3 Lipopolysaccharide bilayer interaction with antimicrobial peptides
3.3.1 Morphological characteristics of LPS bilayers
3.3.2 Lipopolysaccharide bilayer interaction with a cyclic antimicrobial peptide
3.3.3 Lipopolysaccharide bilayer interaction with a linear antimicrobial peptide
3.4 Comparison of the action of a cyclic and a linear peptide on model membranes
4. Antimicrobial peptide action on bacterial biofilms
4.1 Bacterial growth in planktonic form
4.2 Formation of a bacterial biofilm
4.3 Cyclic antimicrobial peptide action on a bacterial biofilm
4.4 Linear antimicrobial peptide action on a bacterial biofilm
4.5 Comparison of the action of a cyclic and a linear peptide on biofilms
5. Conclusions
References