Applications are requested from students and post-docs who wish to
attend a 3.5 day imaging workshop at UCL focusing on confocal
microscopy techniques. The workshop will run from the morning of the
8th June to lunchtime on the 11th June.
Registration and accommodation costs will be covered by the ZF-MODELS
Integrated Project ([Only registered users see links. ]).
Day 1 – lectures
Basics of microscopy: setting up Koehler illumination, Phase contrast,
DIC. Point spread functions and optical resolution. Objectives –
what do all the codes mean? What are the benefits and drawbacks of the
different objectives? Which one should I choose for what job?
Introduction to confocal microscopy: Confocal principle, optical
paths, signal cross talk. Digital images, optical vs digital
resolution, pixels, voxels and grey levels. Digital and physical
regions of interest - scanning and analysis. Optical sectioning and
3D reconstruction and rendering methods.
Live cell imaging confocal and multiphoton confocal microscopy: FRET,
FRAP, FLIP, FLIM. 2 photon concept, advantages and drawbacks. SHG
and THG imaging.
Image analysis and 3D reconstruction: Image composition and
properties. Signal co-localization analysis. Deconvolution
analysis. Application of algorithms to improve image quality –
dangers! 3D reconstruction, volume rendering and visualization.
Comparison of software packages available.
Fluorescent probes and cell labeling: Basics of fluorescence,
luminescence and spectral properties and consideration. Chemical
fluorophores, fluorescent proteins (standard and advanced), FLAsh
ReAsh bi-arsinical probes. Labelling techniques, bathing, AM dyes,
electroportation, transfection microinjection, gene gun.
Day 2 & 3 & 4 – circus of practical classes to rotate round in groups
of 4 (to make sure everyone gets hands on experience).
1. Basic microscope set up and alignment for Koehler illumination,
Phase contrast, DIC. Alignment of Hg lamps. Confocal basics – image
acquisition, correct dynamics range, eliminate cross talk, Z series,
lambda scan, image projection and rotation.
2. Live cell imaging FRAP and FRET techniques.
3. Multiphoton and second harmonic imaging.
4. Image analysis and 3D reconstruction practical.
5. Zebrafish specific module - high-resolution fish brain imaging,
including some detail on transgenics, mounting, imaging,
reconstruction and presentation
Deadline for applications: 10th May 2009
To apply, send a CV & cover letter explaining why you would like to
attend and a letter of support from your lab to Steve Wilson
([Only registered users see links. ].uk)