Table 1.

Measurement and correction of shifts for multi fluorescence cryo microscopy imaging.

Merged images	Axis	Measured shifts [nm]			Alignment precision σA [nm]
		Max	Min	Average	Max	Min	Average
Green/Blue	X	1417	3	142	29	8	17
	Y	357	4	156	30	13	20
Red/Blue	X	3098	8	415	25	5	13
	Y	868	1	309	33	9	15

For each set of images, the shift between red/blue and green/blue fluorescence images was measured and corrected according to TetraSpeck microspheres of 200 nm diameter. The alignment precision σ^A is given in nm. Data corresponding to the maximum, minimum and average values were measured on 8 data sets recorded on plunge-frozen cells infected with Ad5-488 viruses and 8 data sets recorded on a plunge-frozen microsphere sample. A total of 3 to 20 TetraSpeck microspheres were used per image for the calculations.