Light Microscopy for You
From BioDIP
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This brief, crash course, covers: | This brief, crash course, covers: | ||
#What you need to know about light microscopy and the LMF/IPF but never knew to ask. | #What you need to know about light microscopy and the LMF/IPF but never knew to ask. | ||
− | #Concepts you need to know / refresh, including tips and tricks, | + | #Concepts you need to know / refresh, including tips and tricks, concerning how to improve your imaging experiments and impress your RGL/PI, and TAC. |
− | concerning how to improve your imaging experiments and impress your RGL/PI, and TAC. | + | |
#Safety: Lasers, mercury lamps, etc. | #Safety: Lasers, mercury lamps, etc. | ||
#How to use the LMF / IPF | #How to use the LMF / IPF |
Revision as of 11:51, 27 September 2010
Contents |
LM4U - light microscopy, for you!
What is LM4U? - Briefing Abstract
Look, its not a course! OK?! This brief, crash course, covers:
- What you need to know about light microscopy and the LMF/IPF but never knew to ask.
- Concepts you need to know / refresh, including tips and tricks, concerning how to improve your imaging experiments and impress your RGL/PI, and TAC.
- Safety: Lasers, mercury lamps, etc.
- How to use the LMF / IPF
Target Audience: Everyone doing light microscopy- ESPECIALLY NEW-COMERS!!!
Content
(1 slide per point)
Introduce yourself
- Who are we?
- Who are you?
Practical Microscopy Tips and Tricks
- What is a Microscope - Provdides Magnification and more importantly Resolution.
- Basic Ray Optics of a Lens.
- refraction
- Conjugate planes
- Single Lens has 2 focal planes FFP and BFP
- little microscope
- What is confocal
- Resolution
- optical resolution - Point Spread Function / Airy Disk
- Pixel Spacing (size?) Physical CCD dexels vs Picture Elements vs point scanning
- Numerical Aperture vs Magnification : Abbe/Rayleigh Resolution d = lambda / 2NA
- What is contrast? (No light = no image; no contrast = no resolution)
- Fluorescence and Stokes Shift
- Spectrum databases / viewers for planning expt.
- Fluorescence and Stokes Shift
- Detectors: CCD vs. PMT Widefield vs point detection
- Digitization
- Detector Saturation
- Objective language / reading
- Coverslip thickness is part of the objective design (#1.5 0.17 mm +- 0.01)
- Lens Immersion media
- old oil, wrong oil, wrong collar setting
- Lens cleaning
- safety ; mercury lamps, lasers (jewelry, interlocks)
- Project Planning / Workflow
Admin
- Admin intro
- New User 30 min new user checklist