SEPTEMBER 29 and 30, 2005

Coordinators:
José Carrascosa (Spain) and Fredi Sánchez (Venezuela)

Faculty:
José L. Carrascosa
Departamento de Estructura de Macromoléculas. Centro Nacional de Biotecnología. Campus Universidad Autónoma de Madrid, Madrid, España.

José R. Castón
Departamento de Estructura de Macromoléculas. Centro Nacional de Biotecnología, Campus de la Universidad Autónoma de Madrid, Madrid, España

Cristina Risco
Departamento de Estructura de Macromoleculas. Centro Nacional Departamento de Biología Estructural de Biotecnología, Campus Universidad, Lab. Estructura Molecular Autónoma de Madrid- Madrid- España

Fredi Sánchez
Instituto Venezolano de Investigaciones Científicas (IVIC). Caracas, Venezuela

Program:

• Why Cryo-TEM?, vitrified samples, relevance of the use of frozen-hydrated tissues in electron microscopy. Cryo-TEM of biological samples. (F.Sánchez)
• Properties and structure of water molecules. Water in cells and tissues. Freezing of bioplogical tissues and its effect in biological membranes. Principles of freezing. Structure of water under vacuum. Different methods to avoid intracellular ice formation. Use of cryo-protectants. Fast freezing of biological samples. (F.Sánchez).
• Freeze-fracture, freeze-substitution and cryo-sections. Freeze-fracture and metal replica using different metals. Cryo-substitution protocols for structural analysis and macromolecular detection. Cryo-sections for immuno cytochemistry. Three-dimensional studies. (C. Risco).
• Cryo-microscopy of isolated particles. Sample preparation and freezing procedures. Visualization and digital processing. Classification, two-dimensional averaging and three-dimensional reconstruction. (J.L. Carrascosa)
• Three-dimensional reconstruction of particles with symmetries. Assymmetric particle reconstruction. Mapping specific structural components. Tomographic reconstruction. (J.L. Carrascosa).
• Cryo-micoscopy of virus. Methods for the three-dimensional reconstruction of icosahedral viruses. Requirements for high resolution structure determination. (J.R. Castón).
  

• Alfredo Feria-Velasco. Historical aspects of confocal microscopy
• Michael Koberg. Basis of the confocal microscopy
• Raul Mena López. Specimen preparation for confocal microscopy

Confocal microscopy applications will be discussed during the “Symposium on Application of Confocal Microscopy” that will be part of the Scientific Program of the Congress.

• Alwyn Eades (USA). E.mail:jae5@lehig.edu
• Luis Bejar (Mexico). E-mail: lbgomez@zeus.umich.mx

Advances in electron sources and lenses
Focused Ion Beam (FIB) instruments
Electron backscattering diffraction
Monte Carlo simulation
Nanoparticle characterization
Energy dispersive spectroscopy (EDS) and energy loss spectroscopy (EELS)
In situ microscopy
Correction of lens aberration and prospects for the future

• SEPTEMBER 29, 2005
  
  Coordinator: John Mansfield (EE.UU) jfmjfm@umich.edu
• There has been an explosion of interest in remote microscopy. I have seen use in collaborative research and also teaching. Itallows users access to state-of-the-art facilities that they themselves cannot afford. Several laboratories have assembled remote control systems, often at a great cost in hardware and computer programming. low cost alternatives are welcome. This course will illustrate how it is possible to achieve remote control of an instrument with reasonably cheap computer equipment (~$1000), free software and a moderately fast Internet connection (Cable modem or DSL). Instruments that are capable of being remotely controlled in this manner include several from JEOL, FEI and Hitachi. Complete details of how to set up a remote control system and what hardware and software to install will be discussed.
  

• SEPTEMBER 29, 2005
  
• Coordinator: Rudolf Reichelt (Germany). reichru@uni-muenster.de
  
• Faculty:
  Rudolf Reichelt and Werner Stracke (W.Stracke@uni-muenster.de)
  Institute for Medical Physics and Biophysics, University of MuensterRobert-Koch-Str. 31, D-48149 Muenster, Germany.
  
• Outline:
  The theoretical part given in advance by Prof. R. Reichelt.
  
  The objectives of the experimental part of the AFM course are both to illustrate the transfer of the acquired basics to the function of the AFM and to demonstrate the influence of the parameter settings “with the physics behind it” on the quality of the images. Therefore, the basic handling and operating an AFM is demonstrated focusing on data acquisition, data processing and presentation of data. Mainly it is dealt with different imaging modes; however, the operation in force spectroscopy mode is briefly discussed as well.
  
  Introduction:
  o Schematic presentation of the operation of an AFM, e.g. identifying the microscope setup and getting to know the controller software
  o Instruction in the basic handling, e.g. of different modes of data acquisition
  o Work with a specimen: How to start and what to do? e.g. sample preparation, choice of operation mode
  
  Experimental Work:
  o How to acquire data? e.g. proceeding for improving the signal-to-noise ratio, proceeding for confirming the image reality
  o How to evaluate data? e.g. distinguishing between image- and instrumentation artifacts, analyzing the raw data quantitatively
  o How to process data? e.g. image-flattening and –filtering, usage of image processing/analyzing tools
  o How to present data? e.g. enhancing the desired features, generating height profile diagrams
  
• The details of the hands on work in the course depend on the particular type of AFM locally available. The experimental part of the course will be illustrated by a step by step computer presentation.