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From Wednesday, 9 November 2011 the weekly lectures will be every Wednesday, 8:30

From Wednesday, 9 November 2011 the weekly lectures will be every Wednesday, 8:30 Humboldtstr. 8, Seminar Room 3 (SR 3). 2. Contrast modes in light microscopy: Bright field. Blackboard exercises: Coherent vs. Incoherent imaging. The Concept of a A mplitude S pread F unction.

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From Wednesday, 9 November 2011 the weekly lectures will be every Wednesday, 8:30

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  1. From Wednesday, 9 November 2011 the weekly lectures will beevery Wednesday, 8:30 Humboldtstr. 8, Seminar Room 3 (SR 3)

  2. 2. Contrast modes in light microscopy: Bright field Blackboard exercises:Coherent vs. Incoherent imaging The Concept of a Amplitude Spread Function Image Field as a Convolution of Object with ASF The Concept of a Point Spread Function Imaging as a Convolution of Object with PSF

  3. Fourier-space & Optics

  4. Spatial Frequencies and Fourier Transforms Realspace: Intensity x [m] Fourier space: Amplitude k [1/m]

  5. from:http://www-groups.dcs.st-and.ac.uk/ ~history/PictDisplay/Fourier.html Even better approximation: from:http://members.nbci.com/imehlmir/ Fourier Analysis

  6. Cosine transform Fourier transform

  7. ky ky kx kx Constructing images from waves CorrespondingSine-Wave SpatialFrequency AccumulatedFrequencies SumofWaves

  8. Constructing images from waves CorrespondingSine-Wave SpatialFrequency AccumulatedFrequencies SumofWaves

  9. Real Space (PSF) Reciprocal Space (ATF) Lens Cover Glass ky y x kx Focus z kz Oil Intensity in Focus (PSF)

  10. McCutchen generalised aperture Ewald sphere

  11. IFT A(k) a(r)

  12. a(r) |a|2(r)

  13. IFT A(k) a(r) |.|2 ? ? |a|2(r) FT

  14. ? Fourier Transform ~ ~ ~* I(k) = A(k) A(-k) OTF ATF Epifluorescent PSF I(x) = |A(x)|2 = A(x) · A(x)*

  15. Region of Support kx,y kz Convolution: Drawing with a Brush

  16. ! kx,y kz Optical Transfer Function (OTF)

  17. Missing cone

  18. Objective Lense Tube Lense CCD Back FocalPlane 2. Contrast modes in light microscopy: Bright field Scattering / Absorbtion Bright FieldTransmission Dark object on bight backgroundRelative scattering angle and wavelength defines resolutionCondensor AND objective Numerical Aperture matterContrast decreases when resolution increases

  19. kout Kobj kin 2. Contrast modes in light microscopy: Bright field Interference of diffracted lightwith the undiffracted reference (first Born approx.) Range ofDetection Angles "Bragg condition" Holgraphy with plane wave illumination: infinitely little 3D information is acquired!

  20. 2. Contrast modes in light microscopy: Bright field 1.? 2.? 3.? 6.? 4.? 5.? http://biology.about.com

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