- #1
u0362565
- 52
- 0
Hi all,
If a camera images a fluorescent molecule gaussian function with diameter roughly 300nm and each image pixel represents 160x160nm how could you say with higher precision where the molecule is located within that pixel. For instance if the localisation precision turns out to be 40nm how is it possible to derive this. I know it's to do with counting the number of photons collected and this given an independent measurement of the molecules position but surely only to within the resolution of the pixel size 160x160nm so how is it that the precision of localisation could be 40nm!? This is the basis of single molecule localisation microscopy but I can't figure out between the optics of the microscope & the engineering of the ccd how these resolution figures are possible. I'm really missing something here.
If a camera images a fluorescent molecule gaussian function with diameter roughly 300nm and each image pixel represents 160x160nm how could you say with higher precision where the molecule is located within that pixel. For instance if the localisation precision turns out to be 40nm how is it possible to derive this. I know it's to do with counting the number of photons collected and this given an independent measurement of the molecules position but surely only to within the resolution of the pixel size 160x160nm so how is it that the precision of localisation could be 40nm!? This is the basis of single molecule localisation microscopy but I can't figure out between the optics of the microscope & the engineering of the ccd how these resolution figures are possible. I'm really missing something here.