3. Mold design with six circular rings of 25 mm in diameter and
200 μm in height (supplier: http://ufluidix.com).
4. Glass wafer.
5. Plastic-tipped tweezers.
2.5.3 InfectChip (See
Note 4)
1. Micro-patterned coverslip (Fig. 1).
2. Agarose Sheet: see Subheading 3.5, steps 7–12 for preparation.
3. Microfluidic chip in PDMS with micropatterned serpentine
channel.
4. Plastic-tipped tweezers.
5. Holder consisting of a polymethyl methacrylate (PMMA)
block (5 mm thick) and an aluminium plate with a 14 mm
central hole.
6. Six plastic screws.
7. 35 mm cell culture dishes, tissue culture-treated polystyrene (Falcon 353001).
8. Syringe 50 mL luer lock (BD 300865).
9. HL5c medium including glucose supplemented with vitamins
and microelements (ForMedium): resuspend 26.55 g of powder in 1 L of deionized water. Filter-sterilize (see Note 1).
10. Beaker 200 mL for the HL5c medium trash and then renumber accordingly.
11. Parafilm.
12. Spinning disc confocal microscope: Intelligent Imaging Innovations Marianas SDC mounted on an inverted microscope
(Leica DMIRE2), 63Â Numerical Aperture 1.4 objective (see
Note 5).
2.6 High-Content
Microscopy
1. Ibidi μ-plates 96-well black (89626).
2. Gas semipermeable films (4titude Moisture Barrier Seals 96).
3. High-content microscope: ImageXpress Micro Confocal
IXM-Molecular Devices using a 40Â plan Apo N.A. 0.95
objective.
2.7 Developmental
Cycle
1. 8-Well coverslip “μ-slide 8 Well Glass Bottom” by Ibidi
(80826).
2. Sorensen–Magnesium–Calcium (SorMC) buffer: Autoclaved
15 mM KH 2 PO 4 , 2 mM Na 2 HPO 4 , 50 μM, MgCl 2 , 50 μM
CaCl 2 , nominal pH 6.0.
3. FACS SH-800 device (Sony™): This system provides a high
level of automation and ease of use available in a cell sorter. The
automation includes side stream calibration, drop delay calibration, sort monitoring, and droplet calibration.
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200 μm in height (supplier: http://ufluidix.com).
4. Glass wafer.
5. Plastic-tipped tweezers.
2.5.3 InfectChip (See
Note 4)
1. Micro-patterned coverslip (Fig. 1).
2. Agarose Sheet: see Subheading 3.5, steps 7–12 for preparation.
3. Microfluidic chip in PDMS with micropatterned serpentine
channel.
4. Plastic-tipped tweezers.
5. Holder consisting of a polymethyl methacrylate (PMMA)
block (5 mm thick) and an aluminium plate with a 14 mm
central hole.
6. Six plastic screws.
7. 35 mm cell culture dishes, tissue culture-treated polystyrene (Falcon 353001).
8. Syringe 50 mL luer lock (BD 300865).
9. HL5c medium including glucose supplemented with vitamins
and microelements (ForMedium): resuspend 26.55 g of powder in 1 L of deionized water. Filter-sterilize (see Note 1).
10. Beaker 200 mL for the HL5c medium trash and then renumber accordingly.
11. Parafilm.
12. Spinning disc confocal microscope: Intelligent Imaging Innovations Marianas SDC mounted on an inverted microscope
(Leica DMIRE2), 63Â Numerical Aperture 1.4 objective (see
Note 5).
2.6 High-Content
Microscopy
1. Ibidi μ-plates 96-well black (89626).
2. Gas semipermeable films (4titude Moisture Barrier Seals 96).
3. High-content microscope: ImageXpress Micro Confocal
IXM-Molecular Devices using a 40Â plan Apo N.A. 0.95
objective.
2.7 Developmental
Cycle
1. 8-Well coverslip “μ-slide 8 Well Glass Bottom” by Ibidi
(80826).
2. Sorensen–Magnesium–Calcium (SorMC) buffer: Autoclaved
15 mM KH 2 PO 4 , 2 mM Na 2 HPO 4 , 50 μM, MgCl 2 , 50 μM
CaCl 2 , nominal pH 6.0.
3. FACS SH-800 device (Sony™): This system provides a high
level of automation and ease of use available in a cell sorter. The
automation includes side stream calibration, drop delay calibration, sort monitoring, and droplet calibration.
188
Manon Mottet et al.