Light microscopy Lysmikroskopi

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Presentation transcript:

Light microscopy Lysmikroskopi Optical microscopy MENA3100,OBK, 28.01.15

Only draw the ones that are ”important”

One lense Thin lense Projection

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

Focal plane

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

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

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

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

s (= v) s’ f f f

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

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

Fourier transformation Read the text from Johan Taftø

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

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

Magnifying glass Antony van Leeuwenhoek (1632−1723)

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

Stereo mikroskope

Compound microscope for transmitted light Microscope for transmitted light

Mikroskope for reflected light

s (= v) s’ f f f

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

Illuminating system Prøve Kondensorlinse Aperturblender Aperture diaphragm Feltblender Field diaphragm Filament

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

Avbildningssystemet Eyepiece Okular ”Tube lens” Objektiv Prøve

Primært bilde

Snell’s law: = Refraction Refraksjon ─ lysbrytning 1 2 Sin1 Sin2 v1 v2 =

Chromatic aberation

Spherical aberasjon + astigmatisme, koma, fortegning ...

Correcsions Objective 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

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

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

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

Numerical aperture (NA) NA = nsina Resolution Oppløsning R = 0,61l/NA

Objectives for oil immersion

Optical anisotropy Calcite

Optically active crystals in the microscope

Optically active crystals in the microscope

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