howto:check_overlap_of_different_color_confocal_volumes
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| howto:check_overlap_of_different_color_confocal_volumes [2013/11/27 10:04] – admin | howto:check_overlap_of_different_color_confocal_volumes [2015/02/25 13:55] (current) – bornemann | ||
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| - | ====== How to check the overlap | + | {{tag> |
| + | ====== How to Check the Overlap | ||
| Because both lasers are fed through the same fiber, there cannot be any optical beam displacement. According to my records, your system has the latest apochromatic fiber outcoupler, therefore there is practically nothing you can improve: lasers mounted in the [[products: | Because both lasers are fed through the same fiber, there cannot be any optical beam displacement. According to my records, your system has the latest apochromatic fiber outcoupler, therefore there is practically nothing you can improve: lasers mounted in the [[products: | ||
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| You need an appropriate dual band major dichroic and PIE excitation with the Sepia. In this case there is no need for detection color splitting. One detector is sufficient. Use a single long pass filter, target and image a single bead. Of course, you have to set the laser intensities so that both laser excitations contribute with approximately half of the final detected sum count rate. | You need an appropriate dual band major dichroic and PIE excitation with the Sepia. In this case there is no need for detection color splitting. One detector is sufficient. Use a single long pass filter, target and image a single bead. Of course, you have to set the laser intensities so that both laser excitations contribute with approximately half of the final detected sum count rate. | ||
| - | In [[Products:SymPhoTime]], calculate two images: one for each PIE time window, that is, one for each excitation laser. (It is easy with the [[Products:SymPhoTime]], use the graphical tool for setting the time-gates.) By comparing the two images, you should see no significant displacement. | + | In [[Products:SymPhoTime64]], calculate two images: one for each PIE time window, that is, one for each excitation laser. (It is easy with the [[Products:SymPhoTime64]], use the graphical tool for setting the time-gates.) By comparing the two images, you should see no significant displacement. |
howto/check_overlap_of_different_color_confocal_volumes.txt · Last modified: 2015/02/25 13:55 by bornemann