BasicsCoursePhDprog-PeterEvennetApril2013
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Contents |
PhD Program: Basics of Light Microscopy - Peter Evennett / LMF - Course April 2013
Setup -Fri 05 April
Setup of all needed course material in Small Auditorium
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 (8th April)
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 | 11:05 | Session 1 Theory |
PRINCIPLES OF LIGHT MICROSCOPY Historical aspects Very basic components and principles ( incl. Refraction-basics) |
PETER EVENNETT (JAN) | Jan shortly showing airy pattern from hole in mirror at one of the teaching microsopes (+ screen) |
11:05 | 11:15 | BREAK | BREAK | ||
11:15 | 11:50 | SESSION 1 Practice SHORT (5min !!!) Rotation |
capillary/plastic (Huisken) in oil demo Coin-in-cup demo red and green laser through milky water with black background |
(PETE)Jan PETER BRITTA (SILKE) |
*capillaries, oil * coin-in-cup, water *laser pointer, glass with "milky" water and black background |
11:50 | 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 (Davide will watch the time) |
Milky Glass Block demo Show and discuss microscope parts (upright and inverted) NA measurements |
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 Epi 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 (9th April)
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 & Practice |
Eyepieces 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" |
10:45 | 11:00 | BREAK | COFFEE 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 | 14:15 | SESSION 2 Demo and Theory |
Dan's diffraction setup demonstration | SILKE | Dan's ABBE Diffraction demo setup |
14:15 | 15:00 | SESSION 3 Theory and Demo |
Lens aberrations and corrections | PETER EVENNETT JAN |
magnetic lens + laser suitcase on white board| |
15:00 | 15:30 | SESSION 4 Theory |
Introduction to contrast | PETER EVENNETT | |
15:30 | 15:45 | BREAK | COFFEE BREAK | ||
15:45 | 16:05 | SESSION 4a Theory |
Dark Field microscopy | PETER EVENNETT | |
16:05 | 16:30 | SESSION 4a Practice |
Dark Field microscopy | LMF TEAM | diatome slides |
16:30 | 16:50 | SESSION 4b Theory |
Basics of Phase Contrast microscopy | PETER EVENNETT | |
16:50 | 17:30 | SESSION 4b Practice |
Phase Contrast microscopy | LMF TEAM | cheek cell samples |
17:30 | : | SESSION 4b Theory |
Details of Phase Contrast microscopy | PETER EVENNETT |
DAY THREE (10th April)
FROM | TO | ACTIVITY | TOPICS | Responsible person(s) | Materials needed |
9:00 | 10:00 | Recap | day 2 | LMF TEAM | Questions |
10:00 | 10:30 | SESSION 1 Theory |
Objective reading | PETER EVENNETT LMF TEAM |
objectives, eyepieces, polylux |
10:30 | 10:45 | BREAK | BREAK | BREAK | |
10:45 | 11:45 | SESSION 1 Practice Group rotation |
Cover glasses, sample mounting and Objective cleaning Objective reading |
JAN BRITTA, SILKE |
Coverglasses (High preciscion), different oils, slides, different cleaning reagents, cover glass thickness measure meter at least 7 different, "interesting" objectives (for a group of 6-7) |
12:15 | 12:45 | SESSION 2 Theory |
Polarized light microscopy | PETER EVENNETT | |
12:45 | 13:45 | BREAK | LUNCH | ||
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 | 16:15 | Question round | Questions to Peter Evennett | PETER EVENNETT LMF TEAM |
|
16:15 | 17:50 | SESSION 3 Theory & Practice |
Detectors | BRITTA | • CCD chip • example cameras showing different chip sizes • RGB Slider Spot Camera setup Demo: CCD chip under stereo microscope |
17:50 | 18:15 | SESSION 3 Practice |
PIXELS | LMF TEAM | Sesame seeds, sunflower seeds, nudels, gumibears and a checker board |
DAY FOUR (11th April)
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 | JAN | - | - |
10:45 | 11:00 | BREAK | BREAK | BREAK | BREAK | BREAK |
11:00 | 12:15 | SESSION 2 Theory & Practice |
Fluorescence Microscopy | DAVIDE | • 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 (Convallaria) |
13:00 | 14:00 | BREAK | LUNCH | LUNCH | LUNCH | LUNCH |
14:00 | 15:45 | SESSION 3 Theory & Practice |
What is a pixel Analog to Digital conversion Nuyquist-Shannon Quantitative Imaging Spatial Calibration |
BERT DAVIDE |
- | • pixel size calculation on a given optical 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(Davide, 2 students) N-Storm (Britta, 3 students) p-DV (Bert, 2students) L1 (Pete, 2 students) self-built-syst (Jan, 3 students)) • Fluorescence Sample slides (Kidney / Cells) • camera example sheets |
19:00 | open end | Dinner | with Peter | LMF TEAM |
DAY FIVE (12th April)
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 |
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 | |
15:00 | 15:15 | BREAK | BREAK | BREAK | |
15:15 | 16:00 | SESSION 3 | Course evaluation | LMF TEAM |