Dear Thunderstorm users,
I am a recent postdoc in the university of Michigan and I started to use Thunderstorm for the reconstruction of my dSTORM data. My problem concern the camera settings to set for the reconstruction because in the current system I use the detection element is not an EMCCD camera but a sCMOS camera which is very different . The camera setup parameters asked to use thunderstorm seems to be adapted for EMCCD and not sCMOS. Based on the datasheet associated to my camera I have estimate its photoelectrons per A/D count to 0.49 and the base level (A/D counts) to 1.458 but I am not sure about this numbers because they are very different to the default camera settings. If I ask you this question it is because I have test all of this with my data and I have a correct reconstruction for alpha-tubulin or TOMM20 sub-cellular distributions for example but the sigma distribution of the localized molecules in the sample begin usually after 30nm (very few points below this value). There is some of Thunderstorm users, currently collect data with a sCMOS camera? Could you help me with this issue?
Best Romain
Dear Thunderstorm users, I am a recent postdoc in the university of Michigan and I started to use Thunderstorm for the reconstruction of my dSTORM data. My problem concern the camera settings to set for the reconstruction because in the current system I use the detection element is not an EMCCD camera but a sCMOS camera which is very different . The camera setup parameters asked to use thunderstorm seems to be adapted for EMCCD and not sCMOS. Based on the datasheet associated to my camera I have estimate its photoelectrons per A/D count to 0.49 and the base level (A/D counts) to 1.458 but I am not sure about this numbers because they are very different to the default camera settings. If I ask you this question it is because I have test all of this with my data and I have a correct reconstruction for alpha-tubulin or TOMM20 sub-cellular distributions for example but the sigma distribution of the localized molecules in the sample begin usually after 30nm (very few points below this value). There is some of Thunderstorm users, currently collect data with a sCMOS camera? Could you help me with this issue? Best Romain