BasicsCoursePhDprog-PeterEvennetOctober2012
From BioDIP
(Difference between revisions)
(Created page with '==PhD Program: Basics of Light Microscopy - Peter Evennett / LMF - Course October 2012 == === Setup -Fri 13 April === Setup - all lmf staff move stuff to galleria Equipment nee...') |
(→DAY THREE (10th October): added demo to ccd talk) |
||
(12 intermediate revisions by one user not shown) | |||
Line 1: | Line 1: | ||
==PhD Program: Basics of Light Microscopy - Peter Evennett / LMF - Course October 2012 == | ==PhD Program: Basics of Light Microscopy - Peter Evennett / LMF - Course October 2012 == | ||
− | === Setup -Fri | + | === Setup -Fri 05 October=== |
− | Setup | + | Setup of all needed course material in <span style="color:crimson"> Small Auditorium |
− | Equipment needed | + | <br> |
+ | |||
+ | '''Equipment needed''' | ||
* Microscopes | * Microscopes | ||
* Optical bench | * Optical bench | ||
Line 19: | Line 21: | ||
* 2 polarization filters for each microscope | * 2 polarization filters for each microscope | ||
− | === DAY ONE ( | + | === DAY ONE (8th October)=== |
{| border="4" cellpadding="2" cellspacing="8" {| {{table}} | {| border="4" cellpadding="2" cellspacing="8" {| {{table}} | ||
Line 50: | Line 52: | ||
|- | |- | ||
| style="background:LemonChiffon;"| 11:15|| style="background:LemonChiffon;"| 11:50 | | style="background:LemonChiffon;"| 11:15|| style="background:LemonChiffon;"| 11:50 | ||
− | |align="center" style="background:LemonChiffon;"|SESSION 1<br>Practice<br> SHORT (5min !!!) Rotation|| align="center" style="background:LemonChiffon;"|capillary in oil demo <br>Coin-in-cup demo<br>red and green laser through milky water with black background||align="center" style="background:LemonChiffon;"|PETE<br>PETER<br>SILKE | + | |align="center" style="background:LemonChiffon;"|SESSION 1<br>Practice<br> SHORT (5min !!!) Rotation|| align="center" style="background:LemonChiffon;"|capillary/plastic (Huisken) in oil demo <br>Coin-in-cup demo<br>red and green laser through milky water with black background||align="center" style="background:LemonChiffon;"|(PETE)Jan<br>PETER<br>SILKE (Britta) || align="center" style="background:LemonChiffon;" |*capillaries, oil <br> * coin-in-cup, water <br>*laser pointer, glass with "milky" water and black background |
|- | |- | ||
|- | |- | ||
Line 87: | Line 89: | ||
|} | |} | ||
− | === DAY TWO ( | + | === DAY TWO (9th October)=== |
{| border="4" cellpadding="2" cellspacing="8" {| {{table}} | {| border="4" cellpadding="2" cellspacing="8" {| {{table}} | ||
Line 122: | Line 124: | ||
|- | |- | ||
|- | |- | ||
− | | 13:45||14:15||align="center"| SESSION | + | | 13:45||14:15||align="center"| SESSION 2<br>Demo and Theory || align="center"| Dan's diffraction setup demonstration ||align="center" |SILKE ||align="center" |Dan's ABBE Diffraction demo setup |
|- | |- | ||
|- | |- | ||
− | | 14:15||15:00||align="center"|SESSION | + | | 14:15||15:00||align="center"|SESSION 3<br>Theory and Demo||align="center" | Lens aberrations and corrections ||align="center" |PETER EVENNETT <br> JAN|| align="center" |magnetic lens + laser suitcase on white board| |
|- | |- | ||
|- | |- | ||
− | | 15:00||15:30||align="center"|SESSION | + | | 15:00||15:30||align="center"|SESSION 4<br>Theory||align="center" | Introduction to contrast ||align="center" |PETER EVENNETT || |
|- | |- | ||
|- | |- | ||
Line 137: | Line 139: | ||
|- | |- | ||
|- | |- | ||
− | | 15:45 || 16:05 ||align="center"|SESSION | + | | 15:45 || 16:05 ||align="center"|SESSION 4a<br>Theory|| align="center"| Dark Field microscopy||align="center" |PETER EVENNETT || |
|- | |- | ||
|- | |- | ||
|style="background:LemonChiffon;"| 16:05 | |style="background:LemonChiffon;"| 16:05 | ||
− | |style="background:LemonChiffon;"| 16:30 ||align="center" style="background:LemonChiffon;"|SESSION | + | |style="background:LemonChiffon;"| 16:30 ||align="center" style="background:LemonChiffon;"|SESSION 4a<br>Practice|| align="center" style="background:LemonChiffon;"|Dark Field microscopy||align="center" style="background:LemonChiffon;"|LMF TEAM || align="center" style="background:LemonChiffon;"| diatome slides |
|- | |- | ||
|- | |- | ||
− | | 16:30||16:50||align="center"|SESSION | + | | 16:30||16:50||align="center"|SESSION 4b<br>Theory|| align="center"|Basics of Phase Contrast microscopy ||align="center" |PETER EVENNETT || |
|- | |- | ||
|- | |- | ||
|style="background:LemonChiffon;"| 16:50 | |style="background:LemonChiffon;"| 16:50 | ||
− | |style="background:LemonChiffon;"|17:30||align="center" style="background:LemonChiffon;"|SESSION | + | |style="background:LemonChiffon;"|17:30||align="center" style="background:LemonChiffon;"|SESSION 4b<br>Practice|| align="center" style="background:LemonChiffon;"|Phase Contrast microscopy ||align="center" style="background:LemonChiffon;"|LMF TEAM || align="center" style="background:LemonChiffon;"| cheek cell samples |
|- | |- | ||
|- | |- | ||
− | | 17:30|| : ||align="center"|SESSION | + | | 17:30|| : ||align="center"|SESSION 4b<br>Theory|| align="center"|Details of Phase Contrast microscopy ||align="center" |PETER EVENNETT || |
|} | |} | ||
− | === DAY THREE ( | + | === DAY THREE (10th October)=== |
{| border="4" cellpadding="2" cellspacing="8" {| {{table}} | {| border="4" cellpadding="2" cellspacing="8" {| {{table}} | ||
Line 165: | Line 167: | ||
| align="center" style="background:#f0f0f0;"|'''Materials needed''' | | align="center" style="background:#f0f0f0;"|'''Materials needed''' | ||
|- | |- | ||
− | | 9:00||10:00||align="center"|Recap ||align="center" | day 2 ||align="center" |LMF TEAM || align="center" |Questions | + | | 9:00||10:00||align="center"|Recap ||align="center" | day 2 ||align="center" |LMF TEAM || align="center" |Questions |
|- | |- | ||
| 10:00||10:30||align="center"|SESSION 1<br>Theory||align="center" | Objective reading||align="center" |PETER EVENNETT <br>LMF TEAM || align="center" |objectives, eyepieces, polylux | | 10:00||10:30||align="center"|SESSION 1<br>Theory||align="center" | Objective reading||align="center" |PETER EVENNETT <br>LMF TEAM || align="center" |objectives, eyepieces, polylux | ||
Line 177: | Line 179: | ||
|- | |- | ||
|style="background:LemonChiffon;"| 10:45 | |style="background:LemonChiffon;"| 10:45 | ||
− | |style="background:LemonChiffon;"|11:45||align="center" style="background:LemonChiffon;"|SESSION 1<br>Practice<br>Group rotation||align="center" style="background:LemonChiffon;"| Cover glasses, sample mounting and Objective cleaning<br>Objective reading ||align="center" style="background:LemonChiffon;"|JAN<br>BRITTA, SILKE ||align="center" style="background:LemonChiffon;"| Coverglasses (High preciscion), different oils, slides, different cleaning reagents <br> at least 7 different, "interesting" objectives (for a group of 6-7) | + | |style="background:LemonChiffon;"|11:45||align="center" style="background:LemonChiffon;"|SESSION 1<br>Practice<br>Group rotation||align="center" style="background:LemonChiffon;"| Cover glasses, sample mounting and Objective cleaning<br>Objective reading ||align="center" style="background:LemonChiffon;"|JAN<br>BRITTA, SILKE ||align="center" style="background:LemonChiffon;"| Coverglasses (High preciscion), different oils, slides, different cleaning reagents, cover glass thickness measure meter <br> at least 7 different, "interesting" objectives (for a group of 6-7) |
|- | |- | ||
|- | |- | ||
Line 197: | Line 199: | ||
|- | |- | ||
|style="background:LemonChiffon;"| 15:05 | |style="background:LemonChiffon;"| 15:05 | ||
− | |style="background:LemonChiffon;"|15:40||align="center" style="background:LemonChiffon;"|SESSION 3<br>Practice|| align="center" style="background:LemonChiffon;"|Differential Interference Contrast (DIC)||align="center" style="background:LemonChiffon;"|LMF TEAM|| align="center" style="background:LemonChiffon;"| cheek cell samples | + | |style="background:LemonChiffon;"|15:40||align="center" style="background:LemonChiffon;"|SESSION 3<br>Practice|| align="center" style="background:LemonChiffon;"|Differential Interference Contrast (DIC)||align="center" style="background:LemonChiffon;"|LMF TEAM|| align="center" style="background:LemonChiffon;"| cheek cell samples |
|- | |- | ||
|- | |- | ||
Line 209: | Line 211: | ||
|- | |- | ||
|- | |- | ||
− | | 16:15||17: | + | | 16:15||17:50||align="center"|SESSION 3<br>Theory & Practice|| align="center"|Detectors ||align="center" |BRITTA||align="center" | • CCD chip <br> • example cameras showing different chip sizes <br> • RGB Slider Spot Camera setup <br> Demo: CCD chip under stereo microscope |
|- | |- | ||
|- | |- | ||
− | |style="background:LemonChiffon;"| 17: | + | |style="background:LemonChiffon;"| 17:50 |
− | |style="background:LemonChiffon;"| | + | |style="background:LemonChiffon;"|18:15||align="center" style="background:LemonChiffon;"|SESSION 3<br>Practice|| align="center" style="background:LemonChiffon;"|PIXELS||align="center" style="background:LemonChiffon;"|LMF TEAM ||align="center" style="background:LemonChiffon;"| Sesame seeds, sunflower seeds, nudels, gumibears and a checker board |
− | + | ||
− | + | ||
− | + | ||
− | + | ||
|- | |- | ||
|} | |} | ||
− | === DAY FOUR ( | + | === DAY FOUR (11th October)=== |
{| border="4" cellpadding="2" cellspacing="8" {| {{table}} | {| border="4" cellpadding="2" cellspacing="8" {| {{table}} | ||
Line 235: | Line 233: | ||
| 9:00||10:00||align="center"|Recap ||align="center" | day 3 ||align="center" |LMF TEAM |||| | | 9:00||10:00||align="center"|Recap ||align="center" | day 3 ||align="center" |LMF TEAM |||| | ||
|- | |- | ||
− | | 10:00||10:45||align="center"|SESSION 1<br>Theory||align="center" | Fundamental concepts of fluorescence ||align="center" | | + | | 10:00||10:45||align="center"|SESSION 1<br>Theory||align="center" | Fundamental concepts of fluorescence ||align="center" |JAN||align="center" |- ||align="center" |- |
|- | |- | ||
|- | |- | ||
Line 256: | Line 254: | ||
|- | |- | ||
|- | |- | ||
− | | 14:00||15:45||align="center"|SESSION 3<br>Theory & Practice|| align="center"| | + | | 14:00||15:45||align="center"|SESSION 3<br>Theory & Practice|| align="center"|What is a pixel<br>Analog to Digital conversion<br>Nuyquist-Shannon<br>Quantitative Imaging <br> Spatial Calibration ||align="center" |BERT <br> DAVIDE|||| |
|- | |- | ||
| style="background:mediumseagreen;"|'''15:45''' | | style="background:mediumseagreen;"|'''15:45''' | ||
Line 263: | Line 261: | ||
|align="center" style="background:mediumseagreen;"|'''BREAK'''|| style="background:mediumseagreen;"|''' '''|| style="background:mediumseagreen;"|''' '''|| style="background:mediumseagreen;"|''' ''' | |align="center" style="background:mediumseagreen;"|'''BREAK'''|| style="background:mediumseagreen;"|''' '''|| style="background:mediumseagreen;"|''' '''|| style="background:mediumseagreen;"|''' ''' | ||
|- | |- | ||
− | | 16:00||18:00||align="center"|SESSION 4<br>Practice|| align="center"|Imaging with CCD cameras using LMF systems ||align="center" |LMF TEAM ||align="center"|• Displaying <br>• Pixel size<br>• Look up table<br>• Binning<br>• Exposure time<br>• Saturation<br>• Signal to noise <br>• ROI<br>• Bit depth<br>• Auto map<br>• Auto contrast || align="center"|• Microscopes: <br> DVcore( | + | | 16:00||18:00||align="center"|SESSION 4<br>Practice|| align="center"|Imaging with CCD cameras using LMF systems ||align="center" |LMF TEAM ||align="center"|• Displaying <br>• Pixel size<br>• Look up table<br>• Binning<br>• Exposure time<br>• Saturation<br>• Signal to noise <br>• ROI<br>• Bit depth<br>• Auto map<br>• Auto contrast || align="center"|• Microscopes: <br> DVcore(Davide, 2 students)<br> N-Storm (Britta, 3 students)<br> p-DV (Bert, 2students) <br> L1 (Pete, 2 students)<br> self-built-syst (Jan, 3 students)) <br>• Fluorescence Sample slides (Kidney / Cells) |
|- | |- | ||
|- | |- | ||
Line 273: | Line 271: | ||
'' | '' | ||
− | === DAY FIVE ( | + | === DAY FIVE (12th October)=== |
{| border="4" cellpadding="2" cellspacing="8" {| {{table}} | {| border="4" cellpadding="2" cellspacing="8" {| {{table}} |
Latest revision as of 12:21, 2 November 2012
Contents |
[edit] PhD Program: Basics of Light Microscopy - Peter Evennett / LMF - Course October 2012
[edit] Setup -Fri 05 October
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
[edit] DAY ONE (8th October)
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 SILKE (Britta) |
*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 |
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 |
[edit] DAY TWO (9th October)
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 |
[edit] DAY THREE (10th October)
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 |
[edit] DAY FOUR (11th October)
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 | SILKE | • 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 |
What is a pixel Analog to Digital conversion Nuyquist-Shannon Quantitative Imaging Spatial Calibration |
BERT DAVIDE |
||
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) |
19:00 | open end | Dinner | with Peter | LMF TEAM |
[edit] DAY FIVE (12th October)
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 |