B12, and adjust pH to 7.4. Stock solutions except FeSO 4 and
Tween 80 can be prepared in advance and stored at 4
C for up
to a year. FeSO 4 and Tween 80 solutions should be prepared
immediately prior to albumin supplement preparation. Albumin supplement can be prepared in advance, aliquoted, and
stored between À20
C and + 4
C for up to 18 months (see
Note 4).
4. Complete EMJH medium: 900 mL of EMJH basal salt solution, 100 mL of albumin supplement, pH 7.4. Directly after
preparation, required volumes of complete EMJH medium are
distributed in glass tubes with screw cap that can be stored at
4
C up to 6 months.
2.3 Experimental
Infections
1. Hamsters in individual cages.
2. Petroff-Hausser counting chamber (0.02 mm depth) or similar
counting chamber.
3. Infectious injectate: LiVV at the adequate concentration and
volume (see Subheading 3.1).
4. Control injectate: Liquid EMJH alone (same volume with the
infectious injectate).
5. 1 mL syringes equipped with 25G needle for injection.
6. 70% ethanol dispenser.
7. Gauze.
8. Sharps containers for biohazard material.
2.4 Blood and Tissue
Collection
1. LiVV-infected and non-infected control hamsters.
2. Anesthesia/euthanasia station: CO 2 tank, with pressurereducing valve, liter flow meter, and CO 2 chamber with lid.
3. Dissecting tools: scissors, tweezers, scalpel, table, pins.
4. Beaker with 70% ethanol for decontamination of dissecting
tools.
5. 1 mL syringes and 25G needles (0.5 mm  16 mm) for cardiac
puncture.
6. Sharps containers for biohazard material.
7. 70% ethanol dispenser.
8. MagNA Lyser Green Beads tubes (Roche Life Science). For
DNA extraction from organs, tubes should be filled with
360 μL of lysis buffer from QIAamp DNA Mini Kit (Qiagen;
see Subheading 2.5) prior to tissue collection.
9. EDTA blood collector tubes.
10. RNAlater
® Buffer (Qiagen).
11. PAXgene buffer from PAXgene Blood RNA tube (PreAnalytix,
Qiagen).
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Julie Cagliero et al.
Tween 80 can be prepared in advance and stored at 4
C for up
to a year. FeSO 4 and Tween 80 solutions should be prepared
immediately prior to albumin supplement preparation. Albumin supplement can be prepared in advance, aliquoted, and
stored between À20
C and + 4
C for up to 18 months (see
Note 4).
4. Complete EMJH medium: 900 mL of EMJH basal salt solution, 100 mL of albumin supplement, pH 7.4. Directly after
preparation, required volumes of complete EMJH medium are
distributed in glass tubes with screw cap that can be stored at
4
C up to 6 months.
2.3 Experimental
Infections
1. Hamsters in individual cages.
2. Petroff-Hausser counting chamber (0.02 mm depth) or similar
counting chamber.
3. Infectious injectate: LiVV at the adequate concentration and
volume (see Subheading 3.1).
4. Control injectate: Liquid EMJH alone (same volume with the
infectious injectate).
5. 1 mL syringes equipped with 25G needle for injection.
6. 70% ethanol dispenser.
7. Gauze.
8. Sharps containers for biohazard material.
2.4 Blood and Tissue
Collection
1. LiVV-infected and non-infected control hamsters.
2. Anesthesia/euthanasia station: CO 2 tank, with pressurereducing valve, liter flow meter, and CO 2 chamber with lid.
3. Dissecting tools: scissors, tweezers, scalpel, table, pins.
4. Beaker with 70% ethanol for decontamination of dissecting
tools.
5. 1 mL syringes and 25G needles (0.5 mm  16 mm) for cardiac
puncture.
6. Sharps containers for biohazard material.
7. 70% ethanol dispenser.
8. MagNA Lyser Green Beads tubes (Roche Life Science). For
DNA extraction from organs, tubes should be filled with
360 μL of lysis buffer from QIAamp DNA Mini Kit (Qiagen;
see Subheading 2.5) prior to tissue collection.
9. EDTA blood collector tubes.
10. RNAlater
® Buffer (Qiagen).
11. PAXgene buffer from PAXgene Blood RNA tube (PreAnalytix,
Qiagen).
246
Julie Cagliero et al.