Outline. Phase Contrast why? History. Working Principle. Dark-Field Microscopy. Polarization Microscopy. The Inverted Microscope
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1 Outline Phase Contrast why? History Working Principle Dark-Field Microscopy Polarization Microscopy The Inverted Microscope
2 Phase Contrast Why? Brightfield Microscope: Detection of amplitude objects : Objekt
3 Phase Contrast Why? Brightfield Microscope: Detection of amplitude objects :
4 Phase Contrast Why? Brightfield Microscope: Detection of phase objects : Objekt
5 Phase Contrast Why? Brightfield Microscope: Detection of phase objects : Epithelium cells
6 Phase Contrast Why? Brightfield Microscope: Detection of phase objects : Epithelium cells, aperture diaphragm closed
7 History Frits Zernike ( ): 1930: Invents phase-contrast microscopy, 1941: Industrialization (German Wehrmacht), 1953: Nobel Prize for Physics
8 Working Principle Beam Path:
9 Working Principle Objectives: Ph objective Light annulus! Numbering acc. to position in (turret) condenser Caution: Counting chambers! Caution: Fully open aperture diaphragm!
10 Working Principle Control: Auxiliary Microscope Phase annulus Light annulus WRONG! CORRECT!
11 Working Principle Epithelium cells
12 Working Principle Epithelium cells
13 Darkfield Microscopy Photo: Dr. Jörg Haus, 2007 What makes light rays visible?
14 Darkfield Microscopy Shadows: Geometrical Optics Physics: Light described as particle or plane wavefront, w/o diffraction, light rays = wavefront normals
15 Darkfield Microscopy Shadows: Small Structures, Wave Optics? Physics: Light as spherical wavefront, Huygens principle
16 Darkfield Microscopy Shadows: Small Structures, Wave Optics? Young s double-slit experiment
17 Darkfield Microscopy Comparison Brightfield - Darkfield Objektiv Objektiv Präparatebene Präparatebene Kondensor Kondensor Brightfield Darkfield
18 Darkfield Microscopy Condenser: Annulus vs. Cardioid System
19 Darkfield Microscopy H600 LL HP 100: Optical Components Objective, SPL 100/1.25, with iris diaphragm Condenser, NA 1.4
20 Darkfield Microscopy H600 LL HP 100: Optical Components
21 Darkfield Microscopy H600 LL HP 100: Native Blood Investigations
22 Darkfield Microscopy Competitors: Darkfield for Less???
23 Darkfield Microscopy Competitors: Darkfield for Less??? Hund H600 LL HP 100 Novex B mit DF Tube (Standard) binocular trinocular, 100/100 Eyepieces WF 10x/18 WF 10x/(n. a.) Objectives Achro 10/0.25 Plan 4/0.10 SPL 100/ Oel Plan 10/0.25 Plan S40/0.65 Plan 100/1.25 Oel Darkfield Condenser NA 1.4 NA 1.2 Illumination attached to stand, external, 100 W halogen 150 W halogen Price incl. VAT [ ] 4474, ,95
24 Darkfield Microscopy Competitors: Darkfield for Less??? Novex Hund
25 Darkfield Microscopy Competitors: Darkfield for Less??? Novex: Objective iris diaphragm is closed. No high- resolution DF possible!!!
26 Polarization Microscopy Physics: Light has wave properties Unpolarized lightwave Polarized lightwave Polarization filter (Polarizer) Complete extinction Polarization filter (Analyzer)
27 Polarization Microscopy Physics: Light has wave properties Photo: Dr. Jörg Haus, 2008
28 Polarization Microscopy Birefringence: Polarization Rotation
29 Polarization Microscopy Birefringence: Polarization Rotation Glass ceramics between crossed polarizers, with retardation plate Glass ceramics between crossed polarizers
30 Polarization Microscopy Birefringence: Polarization Rotation H 600 with polarization unit HP I: Analyzer with compensator first-order red Strain-free optics Rotary table with object guide Polarizer
31 Polarization Microscopy Two Observation Techniques: Orthoscopic: Polarizer Specimen Analyzer, observation through eyepieces Conoscopic: Observation in back focal plane of objektive with retractable Bertrand lens or auxiliary microscope, high NA of condenser!
32 Polarization Microscopy Two Observation Techniques: Conoscopic:
33 The Inverted Microscope Dedicated Culture Vessels Petri dish Culture flask Terasaki- (96-well-) plate
34 The Inverted Microscope Applications: In-vitro fertilization (with micromanipulators) Cell-culture examinations Examinations from below
35 Wilovert Standard HF 40 Specifications: Lamp 12 V / 30 W Condenser NA 0.25 w/ aperture diaphragm Objectives with large free working distance: A 4/0.10 A 10/0.25 SPL 20/0.50 (LD) SPL 40/0.60 (LD), correction mount
36 Wilovert Standard PH 40 Specifications: Lamp 12 V / 30 W Combin. cond. NA 0.25 w/ aperture diaphragm Objectives with large free working distance: A 4/0.10 A PH1 10/0.25 SPL PH2 20/0.50 (LD) SPL PH2 40/0.60 (LD), correction mount
37 Wilovert AFL 40 PH Additional specifications: HBO 100 W Filter slide Object guide Objectives with large free working distance: AC 4/0.10 A PH1 10/0.25 SPL 20/0.35 (LD) SPL PH2 20/0.50 (LD) SPL PH2 40/0.60 (LD), correction mount
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