Superresolution microscopy 1 Understanding Light Microscopy Diffraction light
- Slides: 28
Super-resolution microscopy 1
Understanding Light Microscopy Diffraction - light “bends” around an object, especially when the size of the object is close to the wavelength of the light Resolution 2
Understanding Light Microscopy R= 0. 61 n • sin = 0. 61 N. A R ½ of wavelength If wavelength = 400 nm then R = ½ X 400 nm = 200 nm
Super-resolution microscopy Super-resolution techniques allow the capture of images with a higher resolution that overcome the diffraction limit. 4
The use of pinhole enhances the resolution, but we cannot call it super-resolution. 5
Super-resolution microscopy - STED Stimulated emission depletion (STED) 6
Super-resolution microscopy - STED Stimulated emission depletion (STED) 7
Super-resolution microscopy - STED Stimulated emission depletion (STED) 8
Super-resolution microscopy - STED Stimulated emission depletion (STED) 9
Super-resolution microscopy - STED Stimulated emission depletion (STED) 10
Super-resolution microscopy - STED Stimulated emission depletion (STED) 11
Super-resolution microscopy Total internal reflection fluorescence (TIRF) 12
Super-resolution microscopy Total internal reflection fluorescence (TIRF) 13
Super-resolution microscopy Total internal reflection fluorescence (TIRF) microscopy evanescent wave 14
Total internal reflection fluorescence (TIRF) microscopy Selectively excite ONLY the surface regions Increase resolution by increased signal-noise-ratio 15
Total internal reflection fluorescence (TIRF) microscopy tubulin staining Epi-fluorescence TIRF Merge http: //www. microscopyu. com/articles/fluorescence/tirfintro. html 16
Structured Illumination Microscopy (SIM) 17
Structured Illumination Microscopy (SIM) 18
Structured Illumination Microscopy (SIM) 19
Structured Illumination Microscopy (SIM) 20
Structured Illumination Microscopy (SIM) 21
Photo-activated localization microscopy (PALM) 22
Photo-activated localization microscopy (PALM) 23
Zeiss Airy. Scan 24
Zeiss Airy. Scan 25
Zeiss Airy. Scan • 0. 2 Airy Unit pinhole, • pixel-reassignment, • deconvolution 26
Zeiss Airy. Scan 27
Assignment Requirement is listed on our syllabus - https: //www. uwyo. edu/zzhang/ - Due: April 30 th, 2020 28