Is the PIXL UV1 or UV2 capable of working with the 'Sapphire' fluorophore?

The 'Sapphire' fluorophore excites at 399nm and emits at 511nm. UV1 (380-390nm) and UV2 (400-410nm) can be used with a 510nm dichromatic filter to limit emission to the camera. Using a blackout screen and increasing exposure helps capture images. PIXL works best with high cytosolic protein concentrations, like super folded GFP or tagged YDC1. Contact technical support for more details.

Answer:

The 'Sapphire' fluorophore has an excitation peak of 399nm and an emission peak of 511nm so there are a number of factors involved here. 

UV 1 and UV 2 have peak wavelengths of 380-390nm and 400-410nm respectively. 

A dichromatic filter at 510 nm limits the emission to the camera - this may be required to remove the visible range background.

Filters used in the PIXL are standard 50mm filters. More details can be found here : How are filters used in the PIXL?

We would recommend the use of the black-out screen, inserted into the internal lip of the door - this will help with exposure when capturing the image. 

In theory with UV2 and the blackout screen installed, and a 510 nm filter in place - this will work.

We have used super folded GFP in our lab that has an emission wavelength of 510nm, with good results. If you don't see anything at first, try leaving all the live imaging settings at default and increasing the exposure setting only. 

A possible downside is protein concentration; the PIXL works well when you have lots of cytosolic fluorescent protein - working on whole-cell fluorescence rather than tagging or subcellular work. As an example in our lab we tagged fluorophores to YDC1 (YLR044C), which gives us a relatively high internal contraction of around 1.84E-30 nM.

If you require more information please contact [e-mail] for assistance.