Thanks for the video. Two questions why do you set your detector range for your HyD4 (Alexa555) to 570-620 nm and dont consider 560, where the extrema emission is? And you said the optimal pixel size is half the resolution, where do I see my resolution in the manual? Thank you in advance
Discussion was not anticipated here, but short answer is that this profile general enough to accommodate various red dyes (not just AF555) and a good starting point for advanced/willing users to tweak. The wavelengths and detectors ranges are chosen for best separation of the four "standard" dyes. Also in practice AF555 is quite bright and you collect plenty of signal in this range. To give a perspective, with 50-100% gain on the HyD and 1-2% laser power you can easily saturate the detector with well stained sample. I am not sure what manual you are referring, but this reference maybe useful to look at: micro.magnet.fsu.edu/primer/java/digitalimaging/processing/samplefrequency/index.html
Very informative and important for all new Leica SP8 users! Thank you so much!
This video was a life saver. Thank you!
That was very comprehensive!
Thanks for the video. Two questions why do you set your detector range for your HyD4 (Alexa555) to 570-620 nm and dont consider 560, where the extrema emission is? And you said the optimal pixel size is half the resolution, where do I see my resolution in the manual? Thank you in advance
Discussion was not anticipated here, but short answer is that this profile general enough to accommodate various red dyes (not just AF555) and a good starting point for advanced/willing users to tweak. The wavelengths and detectors ranges are chosen for best separation of the four "standard" dyes. Also in practice AF555 is quite bright and you collect plenty of signal in this range. To give a perspective, with 50-100% gain on the HyD and 1-2% laser power you can easily saturate the detector with well stained sample. I am not sure what manual you are referring, but this reference maybe useful to look at: micro.magnet.fsu.edu/primer/java/digitalimaging/processing/samplefrequency/index.html
great video, thanks!
very informative. thanks for sharing
这显微镜好高级啊，没用过
Thank you.
THANKYOU THANKYOU THANKYOU
so much money for unsharp images......a DIC with photostacking gives better results