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Light microscopy Lysmikroskopi MENA3100,OBK, 28.01.15 Optical microscopy.

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Presentation on theme: "Light microscopy Lysmikroskopi MENA3100,OBK, 28.01.15 Optical microscopy."— Presentation transcript:

1 Light microscopy Lysmikroskopi MENA3100,OBK, 28.01.15 Optical microscopy

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5 Only draw the ones that are ”important”

6 One lense Thin lense Projection

7 Optical axis Through the centre is not deflected Parallell to the optical axis goes through back focal plane f (Through focal plane before lense becomes parallel optical axis.) f Projected onto a screen Distance one to two focal lengths OBS! The image of the pencil has of cause no width in the direction parallell the optical axis!

8 Focal plane

9 Optical axis ff s (= v)s’s’ M = s’s’ s f v − f f

10 s (= v)s’s’ f 2 f > s > f Magnified, real image ff

11 s (= v)s’s’ f s > 2 f Demagnified, real image ff

12 s (= v)s’s’ f s < f Magnefied, virtual image ff Magnifying glass (forstørrelsesglass, lupe)

13 s (= v)s’s’ f ff

14 Compound microscope Det sammensatte mikroskop Objective Eyepiece Okular Object Real image (Primary image) Real image on retina Virtuell image Usually corrected to «infinite» tubus OBS! Error in figure

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16 All rays originated from the same point are collected here Image plane All rays spread in the same direction are collected here Diffraction plane Fourier plane

17 Fourier transformation Read the text from Johan Taftø

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19 Light microscope We want to see more details Magnification (Forstørrelse) Resolution (Oppløsningsevne) Contrast Optical microscope

20 Simple microscope Magnifying glass (forstørrelsesglass, lupe) Virtuell image

21 Magnifying glass Antony van Leeuwenhoek (1632−1723)

22 Det sammensatte mikroskop ca. 1595 Hans Janssen Robert Hooke (1635−1703) Micrographia, 1664 The compound microscope

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24 Stereo mikroskope

25 Compound microscope for transmitted light Microscope for transmitted light

26 Mikroskope for reflected light

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28 ff s (= v)s’s’ f

29 www.microscopy-analysis.com/files/ jwiley_microscopy/2006_Sept_Hammond.pdf

30 Illuminating system Prøve Filament Feltblender Field diaphragm Aperturblender Aperture diaphragm Kondensorlinse

31 Belysningssystemet Prøve Filament Feltblender Aperturblender Kondensorlinse Köhler-belysning Jevn belysning av prøven Feltblenderen Bestemmer det belyste området Aperturblenderen Bestemmer vinkelen på lyskjeglen som treffer prøven August Köhler 1866─1948

32 Avbildningssystemet Prøve Objektiv ”Tube lens” Eyepiece Okular

33 Primært bilde

34 Refraction Refraksjon ─ lysbrytning Snell’s law: Sin  1 Sin  2 n2n2 n1n1 v1v1 v2v2 == 11 22

35 Chromatic aberation

36 Spherical aberasjon + astigmatisme, koma, fortegning...

37 CorrecsionsObjective Acromate: Chromatic for red and blue, spherical for green Semi-acromate (fluorite) Apochromate: Chromatic for red, green, blue and dark blue, spherical for green and blue

38 Kondensorsystem Eyepice Okular (ocular) More are needed than just the objective

39 The limiting factor: Diffraction R = 0,61 /NA Resolution (Oppløsning)Rayleigh criterion

40 Abbe: ”A microscope image is the interferense effect of a diffraction phenomena”

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42 Numerical aperture (NA) NA = nsin  R = 0,61 /NA Resolution Oppløsning

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44 Objectives for oil immersion

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46 Optical anisotropy Calcite

47 Optically active crystals in the microscope

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49 Remember that geologists often call their book on microscopy on thin sections for optical crystallography, and microscopy with reflected light is ore microscopy (malmmikroskopi). Some web sites: http://www.olympusmicro.com/primer/ http://www.microscopyu.com/ http://www.micro.magnet.fsu.edu/ http://www.micro.magnet.fsu.edu/primer/pdfs/basicsandbeyond.pdf


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