BasicsCoursePhDprog-PeterEvennetOct2011-observed timing and needed materials

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PhD Program: Basics of Light Microscopy - Peter Evennett / LMF - Course May 2011

Setup -Fri 21 Oct

Setup - all lmf staff move stuff to galleria Equipment needed

  • Microscopes
  • Optical bench
  • Spinning disk gadget
  • Polarazing sheets
  • Diffraction Grids
  • Fluorescent samples
  • Fluorescent Beads (Prepare slides)
  • First day Handout for students
  • Light torches
  • cameras and monitors
  • Cleaning stations for each table
  • condenser screwdrivers for each microsocpe
  • oil/DIC objectives for each microscope (next to stand)
  • 2 polarization filters for each microscope

DAY ONE (24th Oct)

FROM TO ACTIVITY TOPICS Responsible person(s) Materials needed
9:00 09:30 FINAL PREP Test everything works LMF
9:40 10:25 INTRODUCTION COURSE INTRO
WHO is WHO? Round table
LMF TEAM White board and pen
10:25 10:55 Session 1
Theory
PRINCIPLES OF LIGHT MICROSCOPY
Historical aspects
Very basic components and principles ( incl. Refraction-basics)
PETER EVENNETT
10:55 11:10 BREAK BREAK
11:15 11:30 SESSION 1
Practice
SHORT (5min !!!) Rotation
capillary in oil demo
Coin-in-cup demo
red and green laser through milky water with black background
DAN
PETER
PETE & JAN
*capillaries, oil
* coin-in-cup, water
*laser pointer, glass with "milky" water and black background
11:30 12:45 SESSION 1
Theory
Introduction to lenses (Lens demo fun)
Lens aberrations (see colour fringes of lighting image with simple lens)
Resolution, Numerical aperture
Magnification
PETER EVENNETT little lenses
12:45 13:45 LUNCH LUNCH
13:45 14:45 SESSION 1
Practice
15min (!!!) Rotation
Milky Glass Block demo
Show and discuss microscope parts (upright and inverted)
NA measurements
DAN
JAN & BRITTA
SILKE
*milky glass block, teaching microscope, coloured filters for microscope, iris objective
*slides with arrows, simple upright and inverted microscope
*horizontal protractor on stand, objective holder, 2 air objective with different (!) NAs (and maybe an iris objective), calculator, pencils, rubber
14:45 15:30 SESSION 2
Theory
Microscope illumination
Conjugate planes
PETER EVENNETT
15:30 15:45 BREAK BREAK
15:50 17:05 SESSION 2
Practice
(30min Rotation)
conjugate planes on Peters microscope
conjugate planes on optical bench
PETER EVENNETT
JAN
*Peters microscope setup
*optical bench demonstrating a transmitted light microscope, incl. light source
17:05 17:25 SESSION 3
Theory
Koehler Illumination
(Microscope alignment)
PETER EVENNETT
17:15 18:30 SESSION 3
Practice
Koehler illumination
(Microscope alignment)
LMF TEAM students manual microscopes, screwdrivers, nice brightfield sample

DAY TWO (25th Oct)

FROM TO ACTIVITY TOPICS Responsible person(s) Materials needed
9:00 10:00 Recap day 1 LMF TEAM Questions
10:00 10:45 SESSION 1
Theory
Epi-Illumination
Lens aberrations and corrections
PETER EVENNETT
10:45 11:00 BREAK COFFEE BREAK Note: have break earlier next time, right after diffraction-string-show
(should be shortly before 11am)
then do all the explanation of Abbe's Diffraction Exp after break
11:00 12:45 Session 2
Theory & DEMONSTRATION
Diffraction and Diffraction slide fun
Diffraction string show
ABBE'S DIFFRACTION EXPERIMENT
Peter's Video (50min)
PETER EVENNETT Diffraction slides, torch, "dashed" string
12:45 13:45 BREAK LUNCH
13:45 14:35 SESSION 3
Demo and Theory
Dan's setup demonstration DAN Dan's ABBE Diffraction demo setup
14:35 15:00 SESSION 1
Theory & Practice
Depth of Field
Depth of Focus
PETER EVENNETT
LMF TEAM
students microscopes with 10x/0.25 and 40x/0.65 objectives
Stereoscope with a "sample"
15:00 15:20 BREAK COFFEE BREAK
15:20 15:55 SESSION 4
Theory
Dark Field microscopy PETER EVENNETT
15:55 16:30 SESSION 4
Practice
Dark Field microscopy LMF TEAM diatome slides
16:30 16:50 SESSION 5
Theory
Basics of Phase Contrast microscopy PETER EVENNETT
16:50 17:30 SESSION 5
Practice
Phase Contrast microscopy LMF TEAM cheek cell samples
17:30  : SESSION 5
Theory
Details of Phase Contrast microscopy PETER EVENNETT

DAY THREE (26th Oct)

FROM TO ACTIVITY TOPICS Responsible person(s) Materials needed
9:00 10:00 Recap day 2 LMF TEAM Questions
10:00 10:45 SESSION 1
Theory
Objective reading
eye pieces
PETER EVENNETT
LMF TEAM
objectives, eyepieces, polylux
10:45 11:00 BREAK BREAK BREAK
11:00 12:30 SESSION 1
Practice
Group rotation (30min. each!)
Cover glasses and sample mounting
Objective reading
Objective cleaning
Dan
Britta, Silke
Jan
Coverglasses (High preciscion), different oils
8-10 different, "interesting" objectives (for a group of 4-5)
slides, oils, different cleaning reagents
12:30 13:30 BREAK LUNCH
13:30 14:00 SESSION 2
Theory
Polarized light microscopy PETER EVENNETT
14:00 14:40 SESSION 2
Practice
Polarized light microscopy LMF TEAM Hair, stone samples, benzocain cristals, rulers, gummi bears, plastic dishes, ...
14:40 15:05 SESSION 3
Theory
Differential Interference Contrast (DIC) PETER EVENNETT
15:05 15:40 SESSION 3
Practice
Differential Interference Contrast (DIC) LMF TEAM cheek cell samples
15:40 16:00 BREAK BREAK BREAK
16:00 17:10 SESSION 4a
Theory & Demo
What is a pixel
Analog to Digital conversion
Nuyquist-Shannon
Dan
17:10 17:30 SESSION 3
Practice
PIXELS LMF TEAM Sesame seeds, sunflower seeds, nudels, gumibears and a checker board
17:30 18:00 SESSION 4b
Theory & Demo
Spatial Calibration and more Dan
18:00 Question round Questions to Peter Evennett PETER EVENNETT
LMF TEAM

DAY FOUR (27th Oct)

FROM TO ACTIVITY TOPICS Responsible person(s) DEMO Activities Materials needed
9:00 10:00 Recap day 3 LMF TEAM
10:00 10:45 SESSION 1
Theory
Fundamental concepts of fluorescence DAN - -
10:45 11:00 BREAK BREAK BREAK BREAK BREAK
11:00 12:15 SESSION 2
Theory & Practice
Fluorescence Microscopy SILKE
LMFTEAM
• Light sources
• Lamp houses
• Spectra
• Filters
• Filter cubes
• Filter spectral charts of each microscope (laminated)
• Spare lamp houses
• Lamps
• Ocean optics equipment
• Four (4) different Filter-sets (cubes), with the appropriate descriptions
• diagramms for drawing filter/fluorophore spectra
12:15 13:00 SESSION 2
Practice
Fluorescence imaging at microscopes LMF TEAM - • Fluorescent labeled samples
13:00 14:00 BREAK LUNCH LUNCH LUNCH LUNCH
14:00 15:45 SESSION 3
Theory & Practice
Imaging with CCD and Quantitative Imaging LMF TEAM
Britta/Dan
• CCD
• Cameras
• CCD chip
*example cameras showing different chip sizes
• RGB Slider Spot Camera setup
15:45 16:00 BREAK BREAK
16:00 18:00 SESSION 4
Practice
Imaging with CCD cameras using LMF systems LMF TEAM • Displaying
• Pixel size
• Look up table
• Binning
• Exposure time
• Saturation
• Signal to noise
• ROI
• Bit depth
• Auto map
• Auto contrast
• Microscopes:
DVcore(Dan, 3 students)
N-Storm (Britta, 3 students)
Inverted CCD (Pete, 2 students)
self-built-syst (Jan, 3 students))
• Fluorescence Sample slides (Kidney / Cells)
19:00 open end Dinner with Peter LMF TEAM

DAY FIVE (28th Oct)

FROM TO ACTIVITY TOPICS Responsible person(s) Materials needed
9:00 10:30 Recap day 4 LMF TEAM Question sheets (!)
10:30 10:45 BREAK BREAK BREAK note: break might have been too early? (usually in between the Optical sectioning talk)
10:45 13:00 SESSION 1
Theory & Demo
Optical sectioning methods
Pros&Cons
LMF TEAM
Dan / Jan
"confocal equipment":
laser pointer, mirror, broken scanning mirror
dual laser pointer (green and red) to demonstrate penetration of diff. wavelength
13:00 14:00 LUNCH LUNCH LUNCH
14:00 15:00 SESSION 2
Small Groups Discussion
What techniques might be good for me
(Own Projects)
LMF TEAM
Dan/Jan/Silke/Britta/Pete?
15:00 15:15 BREAK BREAK BREAK
15:15 16:00 SESSION 3 Course evaluation LMF TEAM
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