REAGENT or RESOURCE	SOURCE	IDENTIFIER
Antibodies
Alexa488 NHS Ester	Thermo Fisher Scientific	Cat#A20000
Alexa647 NHS Ester	Thermo Fisher Scientific	Cat#A20106
Bacterial and Virus Strains
Reovirus type 3 (strain Dearing) (T3D)	Ramig et al., 1977	https://doi.org/10.1128/JVI.22.3.726-733.1977
Chemicals, Peptides, and Recombinant Proteins
Trichlorofluoromethane	Sigma-Aldrich	Cat#48541
Cesium chloride	Sigma-Aldrich	Cat#C4036
Dulbeccos’ modified Eagle medium (DMEM)	Thermo Fischer Scientific	Cat#41966029
Fetal Bovine Serum (FBS)	Capricorn	Cat#FBS-11A
Penicillin/streptomycin	Thermo Fischer Scientific	Cat#15070063
Joklik MEM medium	Sigma-Aldrich	Cat#M0518
L-Glutamine	Thermo Fischer Scientific	Cat#25030081
Neonatal calf serum	Thermo Fischer Scientific	Cat#26010-74
Mitotracker Red FM	Thermo Fischer Scientific	Cat#M22425
Critical Commercial Assays
Zeba Spin Desalting Columns, 7K MWCO	Thermo Fischer Scientific	Cat#89882
Deposited Data
U2OS- reovirus mock Infected (Area 1)	EMPIAR: EMPIAR-10412	BioImage Archive: S-BIAD17
U2OS- reovirus mock Infected (Area 2)	EMPIAR: EMPIAR-10413	BioImage Archive: S-BIAD17
U2OS- reovirus 1hafter infection (Area 1)	EMPIAR: EMPIAR-10414	BioImage Archive: S-BIAD18
U2OS- reovirus 1hafter infection (Area 2)	EMPIAR: EMPIAR-10415	BioImage Archive: S-BIAD18
U2OS- reovirus 2hafter infection (Area 1)	EMPIAR: EMPIAR-10416	BioImage Archive: S-BIAD19
U2OS- reovirus 2hafter infection (Area 2)	EMPIAR: EMPIAR-10417	BioImage Archive: S-BIAD19
U2OS- reovirus 4hafter infection (Area 1)	EMPIAR: EMPIAR-10418	BioImage Archive: S-BIAD20
U2OS- reovirus 4hafter infection (Area 2)	EMPIAR: EMPIAR-10419	BioImage Archive: S-BIAD20
Experimental Models: Cell Lines
L-cells	ATCC	Cat#CRL-6364
U2OS cells	ATCC	ATCC HTB-96
U2OS cells expressing galectin3-mCherry	Maier et al., 2012	https://doi.org/10.1128/JVI.01428-12
BSC-1 cells	ATCC	ATCC CCL-26
Recombinant DNA
eGFP-Rab7 plasmid	Choudhury et al., 2002	Addgene; Cat#12605
Software and Algorithms
SoftWoRX 6.5.2	GE Healthcare	N/A
Chromagnon	Matsuda et al., 2018	https://github.com/macronucleus/chromagnon
Fiji	Schindelin et al., 2012	https://imagej.net/Fiji
SIMcheck	Ball et al., 2015	https://github.com/MicronOxford/SIMcheck
IMOD package (version 4.9.2)	Kremer et al., 1996	https://bio3d.colorado.edu/imod/
eC-CLEM	Paul-Gilloteaux et al., 2017	http://icy.bioimageanalysis.org/plugin/ec-CLEM
Chimera	Pettersen et al., 2004	http://www.cgl.ucsf.edu/chimera/
SurVoS	Luengo et al., 2017	https://diamondlightsource.github.io/SuRVoS/
Linkam’s LINK software	Linkam Scientific	https://www.linkam.co.uk/link-controlsoftware
Cockpit	Micron, Oxford University	https://github.com/MicronOxford/cockpit
Python Microscope	Micron, Oxford University	https://www.python-microscope.org/
Other
TEM grids	Quantifoil	Cat#AU G200F1 finder
Grid holders	Thermo Fisher Scientific	Model#AutoGrid
250 nm gold nanoparticle fiducials	BBI Solutions	Cat#SKU EM.GC250
150 nm gold nanoparticles coated with red Alexa 488	Creative Diagnostics	Cat#GFL-150
PS-Speck Microscope Point Source Kit	Thermo Fisher Scientific	Cat#P7220
CryoSIM 405 nm laser	Omicron-Laserage	Cat#Bluephoton® TA Cat#Deepstar® Series – 375 nm – 488 nm
CryoSIM 488 nm laser	Omicron-Laserage	Cat#Bluephoton® TA Deepstar® Series – 375 nm – 488 nm
CryoSIM 647 nm laser	Omicron-Laserage	Cat#Redphoton® TA Deepstar® Series – 635 nm – 1060 nm
CryoSIM 561 nm laser	Cobolt	Model#Sapphire laser
CryoSIM 520/35 nm filter	Semrock	Cat#BrightLine® single-band bandpass filter, FF01-520/35-25
CryoSIM nematic liquid crystal	Meadowlark Optics	Cat#PDM512
CryoSIM polarization rotator	Meadowlark Optics	Cat#LPR-100-λ
CryoSIM multi band dichroic	Chroma	Cat#ZT405-488-561-647-22.5deg (this paper)
CryoSIM 100X air objective	Nikon	Model#CFI TU Plan Apo EPI 100X, 0.9 NA
CryoSIM cameras	Oxford Instruments	Cat#Andor iXon Ultra 897
CryoSIM & Axioimager Linkam cryostages	Linkam Scientific	Cat#CMS196M LED Cryo Correlative Stage
Axioimager microscope	Carl Zeiss AG	Model#Axio Imager 2
Axioimager 50x objective	Carl Zeiss AG	Model#50x / 0,55 DIC
Confocal microscope	PerkinElmer	N/A
Confocal 60x objective	Nikon	Model#1.49 NA, Apo TIRF
Confocal 100x objective	Nikon	Model#1.4 NA, Plan Apo VC
Confocal camera	Hamamatsu	Cat#CMOS, Orca Flash 4
TXRM microscope	Carl Zeiss X-ray Microscopy, Inc.	Cat#UltraXRM-S220C
TXRM photon detector	Princeton Instruments	Cat#Pixis1024B CCD
TXRM camera	Teledyne	Cat#Retiga 4000R