Fluorescence

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Full Cube assembly Bandpass31001 (Chroma)ExcitationfilterDichroic mirrorEmissionfilterhttp://www.chroma.com/400 500 600 700
Problem 1: cross-excitationa fluorophore is not just excited by wavelength at its peak value, but alsoby wavelength at certain range around the peak, which can extendsinto the area used by other fluorophores.FITC TRITC excitation peaks
Page 1 and 2:
Praktikum zur Fluoreszenz- undKonfo
Page 4 and 5:
Types of Light• Monochromatic•
Page 6 and 7: Lenses, Focus and AberrationsReason
Page 8 and 9: Types of microscopes andIlluminatio
Page 10 and 11: ObjectivesDipping objectives-physio
Page 12 and 13: Resolution• Definition: the small
Page 14 and 15: Resolution!!!!!Magnification identi
Page 16 and 17: Counts for transmitted and reflecte
Page 18 and 19: Depth of FieldThe axial range, thro
Page 20 and 21: Detection Systems Overviewhttp://bi
Page 22 and 23: DetectionDigital Cameras: Photons e
Page 24 and 25: Imaging - Triangle of Frustration©
Page 26 and 27: FluorescenceStokes (1852) - Jablons
Page 28 and 29: Principle of Fluorescence• Molecu
Page 30 and 31: Quantum Efficiency• Only emitted
Page 33: Fluorescent Dyes• Commercially av
Page 36 and 37: Organelle Lights ORGANELLESMitochon
Page 38 and 39: Quantum Dots“Whatever hue want”
Page 40 and 41: IMMUNOFLUORESCENCE-MICROSCOPYIndire
Page 42 and 43: FIXIERUNG:variable-should be suited
Page 44 and 45: PERMEABILISIATION:For antibody diff
Page 46 and 47: • ProLong is good for Alexa dyes
Page 48 and 49: FLUORESCENCE MICROSCOPY
Page 50 and 51: Light Sources / 2Laser• Light amp
Page 52 and 53: Longpassfiltere.g. Longpass 420:Num
Page 54 and 55: Bandpassfiltere.g.: Bandpass 465/70
Page 58 and 59: Problem 2: cross emission (emission
Page 60 and 61: Pseudo-Confocal Microscopy - Struct
Page 62 and 63: Different grids - Differentsectioni
Page 64 and 65: Drawbacks - Troubleshooting• Phot
Page 66 and 67: DeconvolutionLimitations to the res
Page 68 and 69: Literature• J.Pawley, Handbook of
Page 70: Confocal Microscopywhat is differen
Page 73 and 74: Confocal: Functional ElementsDetect
Page 75 and 76: The Pinhole I• Light from below-f
Page 77 and 78: The Pinhole IIIA pinhole of 1 airy
Page 79 and 80: Confocal Overview II• A point lig
Page 81 and 82: Laser II - How are they used?• La
Page 83 and 84: Area Scanning - Point by Point
Page 85 and 86: Optics I: Overview
Page 87 and 88: Optics II: Selecting wavelengthsFil
Page 89 and 90: Confocal Overview IIIno interferenc
Page 91 and 92: Configurations - Zeiss LSM510-71028
Page 93 and 94: Live „Confocal“ - Many Pinholes
Page 95 and 96: Confocal Spinning DiscBeams 1Illumi
Page 97 and 98: Multiphoton MicroscopyPulsed lasers
Page 99 and 100: What can you do?Z- stack including
Page 101 and 102: Z-stack: Reducing Dimensions
Page 103 and 104: Z-stack: Reconstruction• X/Y/Z im
Page 105 and 106: Multicolor Labeling
Page 107 and 108:
Problem II: cross- emissionWhen emi
Page 109 and 110:
The Problems: How to they look like
Page 111 and 112:
Spectral Imaging
Page 113 and 114:
F-techniques - methodology
Page 115 and 116:
iFRAP
Page 117 and 118:
“Optical Highlighters”PHOTOACTI
Page 119 and 120:
Photoactivation (and yet another F-
Page 121 and 122:
Superresolution Ii.e. breaking the
Page 123 and 124:
Superresolution using Fitting Algor
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Fluorescence

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