Slide 1

From Wednesday, 9 November 2011the weekly lectures will be

every Wednesday, 8:30Humboldtstr. 8, Seminar Room 3 (SR 3)

IPC Friedrich-Schiller-Universitt JenaNr.Blackboard exercises: Coherent vs. Incoherent imagingThe Concept of a Point Spread FunctionImaging as a Convolution of Object with PSF2. Contrast modes in light microscopy: Bright fieldThe Concept of a Amplitude Spread FunctionImage Field as a Convolution of Object with ASFIPC Friedrich-Schiller-Universitt JenaNr.2Fourier-space & OpticsIPC Friedrich-Schiller-Universitt JenaNr.k [1/m]Realspace:IntensityAmplitudeSpatial Frequencies and Fourier TransformsFourierspace:x [m]IPC Friedrich-Schiller-Universitt JenaNr.4from: http://members.nbci.com/imehlmir/

Even better approximation:Fourier Analysis

from: http://www-groups.dcs.st-and.ac.uk/~history/PictDisplay/Fourier.htmlIPC Friedrich-Schiller-Universitt JenaNr.5Cosine transformFourier transformIPC Friedrich-Schiller-Universitt JenaNr.

Constructing images from wavesSum of WavesCorrespondingSine-WavekxkykxkyAccumulatedFrequenciesSpatialFrequencyIPC Friedrich-Schiller-Universitt JenaNr.7

Constructing images from wavesSum of WavesCorrespondingSine-WaveAccumulatedFrequenciesSpatialFrequencyIPC Friedrich-Schiller-Universitt JenaNr.8Intensity in Focus (PSF)Reciprocal Space (ATF)kxkzkyReal Space (PSF)xzyLensFocusOilCover GlassIPC Friedrich-Schiller-Universitt JenaNr.

Ewald sphereMcCutchengeneralisedapertureIPC Friedrich-Schiller-Universitt JenaNr.

IFTA(k)a(r)IPC Friedrich-Schiller-Universitt JenaNr.

|a|2(r)a(r)IPC Friedrich-Schiller-Universitt JenaNr.

IFTFT|.|2??|a|2(r)a(r)A(k)IPC Friedrich-Schiller-Universitt JenaNr.Epifluorescent PSF

?I(x) = |A(x)|2 = A(x) A(x)* I(k) = A(k) A(-k) OTFATF~~~*Fourier TransformIPC Friedrich-Schiller-Universitt JenaNr.Convolution: Drawing with a Brushkx,ykzRegion of SupportIPC Friedrich-Schiller-Universitt JenaNr.Optical Transfer Function (OTF)!

kx,ykzIPC Friedrich-Schiller-Universitt JenaNr.

Missing coneIPC Friedrich-Schiller-Universitt JenaNr.Scattering / AbsorbtionBright FieldTransmissionObjectiveLenseTubeLenseBack FocalPlaneDark object on bight backgroundRelative scattering angle and wavelength defines resolutionCondensor AND objective Numerical Aperture matterContrast decreases when resolution increasesCCD2. Contrast modes in light microscopy: Bright fieldIPC Friedrich-Schiller-Universitt JenaNr.18Interference of diffracted lightwith the undiffracted reference (first Born approx.)2. Contrast modes in light microscopy: Bright fieldkinkoutKobjRange ofDetection AnglesHolgraphy with plane wave illumination: infinitely little 3D information is acquired!"Bragg condition"IPC Friedrich-Schiller-Universitt JenaNr.19http://biology.about.com

1.?2.?3.?6.?5.?4.?2. Contrast modes in light microscopy: Bright fieldIPC Friedrich-Schiller-Universitt JenaNr.20