5
Suggestion for “Materials and Methods”
Confocal Z-series stacks (step size 200 nm) were acquired on a Zeiss LSM 880 point
scanning confocal microscope using the Airyscan detector, a 63x Plan-Apochromat 1.4NA DIC
oil immersion objective (Zeiss) and the 405 nm, 488 nm and 561 nm laser lines. The Zeiss Zen
2.3 (black edition) software was used to control the microscope, adjust spectral detection for
the emission of 4
′
,6-diamidino-2-phenylindole (DAPI), Green Fluorescent Protein (GFP) and
tetramethylrhodamine (TMR) SNAP-tag fluorochromes and for processing of the Airyscan raw
images.
Confocal Z-series stacks and tile regions with multiple positions were acquired on a Zeiss
LSM 880 point scanning confocal microscope using photomultiplier tube detectors (PMTs) and
a gallium arsenide phosphide (GaAsP) detector, a 63x Plan-Apochromat 1.4NA DIC oil
immersion objective (Zeiss) and the 405 nm, 488 nm and 561 nm laser lines. The Zeiss Zen 2.3
(black edition) software was used to control the microscope and adjust spectral detection for
the emission of 4
′
,6-diamidino-2-phenylindole (DAPI), Green Fluorescent Protein (GFP) and
tetramethylrhodamine (TMR) SNAP-tag fluorochromes.
Images were acquired on a Zeiss LSM 880 point scanning confocal microscope controlled
with the Zeiss Zen 2.3 (black edition) software, using a 63x Plan-Apochromat 1.4NA DIC oil
immersion objective (Zeiss), the fluorescence filter sets GFP + TX2 and differential interference
contrast (DIC) optics.
Information to be added to the acknowledgements section
This work was partially supported by PPBI - Portuguese Platform of BioImaging (PPBI-
POCI-01-0145-FEDER-022122) co-funded by national funds from OE - "Orçamento de Estado"
and by european funds from FEDER - "Fundo Europeu de Desenvolvimento Regional”.
