ANAT 262 Lecture Notes - Lecture 10: Super-Resolution Microscopy, Super-Resolution Imaging, Evanescent Field
15 Jul 2016
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Document Summary
Use of fluorescent probes (gfp, antibodies, oligonucleotides, chemical stains) In the best case (but not always), the detection limit is a single molecule. Fluorophores excite at short wavelengths and emit at longer wavelengths. Fluorophores can be attached chemically to specific molecules (eg proteins). Gfp can be attached by recombinant dna techniques. Only one point is imaged at any one time. Only point is imaged at any one time. Special optics (pinhole) used to eliminate out-of-focus fluorescence: In focus light focuses inside pinhole and go to detector. Can acquire 3dimensional images: computer can produced images from different planes. Visualize protein distribution in 3-dimensions (perfect for tissues and embryos) Intentionally bleach part of the sample with high energy laser, it causes molecule to react with oxygen, and is no longer fluorescent. Good way to detect if molecules move from different compartments. Only fluorescent part damaged, the molecule is not damaged.
