| National Cancer Institute | Laboratory of Receptor Biology and Gene Expression | National Institutes of Health |
Fluorescence microscopy permits localization of single proteins or observation of cellular organelles within cells. Modern light microscopy is digitally enhanced, thereby facilitating visualization of dim objects, computerized image processing, and quantitative analysis. Although electron microscopy provides better resolution, fluorescence microscopy permits live cell analysis. This field has grown explosively due to development of new fluorophores such as the GFP's, and fast and sensitive imaging systems capable of detecting these markers.
Fluorescence markers have been developed to observe proteins, lipids, DNA and RNA. These include fluorescent antibodies, fluorochrome-labeled oligonucleotides, GFP fusions, and fluorescent stains for lipids, DNA and RNA. Some of these markers are suitable for observations in live cells (e.g. the GFP family when fused to relevant cellular proteins, antibodies when microinjected, certain stains for membranes, and probes for internal pH and Ca2+ concentration).
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| How can one localize YPI in the cell? | In what cellular compartment is YPI localized? Example 1 |
| Is localization of YPI affected by the cell cycle and/or physiological conditions? How can one observe the changes in living cells? Example 2 | |
| What are the exchange and diffusion rates of YPI? How can one observe them in living cells? Example 4 | |
| How can one observe functions of YPI in the cell? | With what cellular proteins is YPI interacting? How can one observe protein co-localization? Example 1 |
| Are mutations of YPI affecting the cell structure? How can one observe microscopically the changes in cellular organelles? Example 3 | |
| How can one develop clinical tests for YPI tissue-specific functions? Example 5 |