Light Microscopy for You
From BioDIP
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| − | = | + | =LM4U - light microscopy, for you!= |
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| − | = | + | ==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: | ||
| + | #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. | # What is a Microscope - Provdides Magnification and more importantly Resolution. | ||
| + | ## Refraction | ||
## Basic Ray Optics of a Lens. | ## Basic Ray Optics of a Lens. | ||
| − | |||
## Conjugate planes | ## Conjugate planes | ||
###Single Lens has 2 focal planes FFP and BFP | ###Single Lens has 2 focal planes FFP and BFP | ||
### little microscope | ### little microscope | ||
### What is confocal | ### What is confocal | ||
| − | # Resolution | + | # Resolution and Diffraction |
## optical resolution - Point Spread Function / Airy Disk | ## optical resolution - Point Spread Function / Airy Disk | ||
##Pixel Spacing (size?) Physical CCD dexels vs Picture Elements vs point scanning | ##Pixel Spacing (size?) Physical CCD dexels vs Picture Elements vs point scanning | ||
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## Digitization | ## Digitization | ||
## Detector Saturation | ## Detector Saturation | ||
| + | ## What is that picture I see on my computer screen? (image, display, LUT, screen calibration) | ||
# Objective language / reading | # Objective language / reading | ||
## Coverslip thickness is part of the objective design (#1.5 0.17 mm +- 0.01) | ## Coverslip thickness is part of the objective design (#1.5 0.17 mm +- 0.01) | ||
| Line 27: | Line 46: | ||
## old oil, wrong oil, wrong collar setting | ## old oil, wrong oil, wrong collar setting | ||
## Lens cleaning | ## Lens cleaning | ||
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# Project Planning / Workflow | # Project Planning / Workflow | ||
| − | # | + | ===Safety=== |
| − | ## | + | # mercury HBO lamps - explosion risk - what to do it it explodes. |
| + | # lasers (jewelry, interlocks, eye level above laser level, dont disable/defeat laser safety devices, dont look at a laser with your remaining good eye. ) | ||
| + | # White light sources (HBO, MetalHalide etc. and Halogen) can also hurt your eyes. | ||
| + | |||
| + | ===Admin=== | ||
| + | |||
| + | # LMF citizenship - policies - local rules: [[Checklist_for_introducing_new_users_to_the_MPI-CBG_LMF/IPF_facility|new user checklist]] | ||
| + | # [[New_user_project_questions]] | ||
Latest revision as of 18:38, 8 March 2011
Contents |
[edit] LM4U - light microscopy, for you!
[edit] 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:
- 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
[edit] Target Audience:
Everyone doing light microscopy- ESPECIALLY NEW-COMERS!!!
[edit] Content
(1 slide per point)
[edit] Introduce yourself
- Who are we?
- Who are you?
[edit] Practical Microscopy Tips and Tricks
- What is a Microscope - Provdides Magnification and more importantly Resolution.
- Refraction
- Basic Ray Optics of a Lens.
- Conjugate planes
- Single Lens has 2 focal planes FFP and BFP
- little microscope
- What is confocal
- Resolution and Diffraction
- 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
- What is that picture I see on my computer screen? (image, display, LUT, screen calibration)
- 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
- Project Planning / Workflow
[edit] Safety
- mercury HBO lamps - explosion risk - what to do it it explodes.
- lasers (jewelry, interlocks, eye level above laser level, dont disable/defeat laser safety devices, dont look at a laser with your remaining good eye. )
- White light sources (HBO, MetalHalide etc. and Halogen) can also hurt your eyes.
[edit] Admin
- LMF citizenship - policies - local rules: new user checklist
- New_user_project_questions
