Light Microscopy for You

LM4U - light microscopy, for you!

What is LM4U? - Briefing Abstract

Look, its not really a taught course! OK?! Its a briefing for new users and whoever else needs to know the bare essentials of light microscopy. This is not a substitute for a full "Basics of light microscopy course", but it will get you started. This very brief, "crash course in light microscopy basics", covers:

1. What you need to know about light microscopy and the LMF/IPF, but never knew to ask.
   1. Concepts you need to know / refresh, including tips and tricks, concerning how to improve your imaging experiments and impress your RGL/PI, and TAC.
   2. Safety: Lasers, mercury lamps, etc.
   3. How to use the LMF / IPF

Target Audience:

Everyone doing light microscopy- ESPECIALLY NEW-COMERS!!!

Content

(1 slide per point)

Introduce yourself

1. Who are we?
2. Who are you?

Practical Microscopy Tips and Tricks

1. What is a Microscope - Provdides Magnification and more importantly Resolution.
   1. Refraction
   2. Basic Ray Optics of a Lens.
   3. Conjugate planes
      1. Single Lens has 2 focal planes FFP and BFP
      2. little microscope
      3. What is confocal
2. Resolution and Diffraction
   1. optical resolution - Point Spread Function / Airy Disk
   2. Pixel Spacing (size?) Physical CCD dexels vs Picture Elements vs point scanning
   3. Numerical Aperture vs Magnification : Abbe/Rayleigh Resolution d = lambda / 2NA
3. What is contrast? (No light = no image; no contrast = no resolution)
   1. Fluorescence and Stokes Shift
      1. Spectrum databases / viewers for planning expt.
4. Detectors: CCD vs. PMT Widefield vs point detection
   1. Digitization
   2. Detector Saturation
   3. What is that picture I see on my computer screen? (image, display, LUT, screen calibration)
5. Objective language / reading
   1. Coverslip thickness is part of the objective design (#1.5 0.17 mm +- 0.01)
   2. Lens Immersion media
   3. old oil, wrong oil, wrong collar setting
   4. Lens cleaning
6. Project Planning / Workflow

Safety

1. mercury HBO lamps - explosion risk - what to do it it explodes.
2. lasers (jewelry, interlocks, eye level above laser level, dont disable/defeat laser safety devices, dont look at a laser with your remaining good eye. )
3. White light sources (HBO, MetalHalide etc. and Halogen) can also hurt your eyes.

Admin

1. LMF citizenship - policies - local rules: new user checklist
2. New_user_project_questions