PDMS Fabrication Protocol¶
- Version: 1.0
- Author: Mingchuan Tang
- Last modified: 2025-10-30
- Purpose: PDMS substrates for on-slide spatial transcriptomics reactions
Required Materials¶
- PDMS: Sylgard 184 Silicone Elastomer Kit (Component A: base, Component B: curing agent)
- Mold: 15 x 15 cm square mold
- Other: Weighing container, stirring rod
Equipment¶
- Balance (0.1g precision or better)
- Vibrator/mixer
- Vacuum pump and desiccator (for degassing)
- Oven (80°C capable)
- Blade or cutting tool
Formulation¶
- Weight ratio: A:B = 10–12 : 1
- Per batch: 120g total
- Standard (10:1): A = 109.1g + B = 10.9g
- Stiffer (12:1): A = 110.8g + B = 9.2g
- Yield: 2 batches (240g) fill 3 molds (15 x 15 cm)
Steps¶
1. Weigh¶
- Weigh Component A (base): 109.1–110.8g (depending on ratio)
- Add Component B (curing agent): 10.9–9.2g
- Total: 120g per batch
2. Mix¶
- Pre-mix with stirring rod
- Place on vibrator for thorough mixing: 5-10 min until uniform
- Bubbles during mixing are normal
3. Degas¶
- Transfer to vacuum desiccator
- Apply vacuum for 15-30 min
- Done when: no visible bubbles rising
- Watch for overflow during degassing
4. Pour¶
- Slowly pour degassed PDMS into mold
- Pour from one side, let liquid self-level
- Pop any remaining small bubbles with a needle
- 2 batches (240g) = 3 molds
5. Cure¶
- Place in oven at 80°C overnight (8-12 h)
- Keep oven temperature stable; avoid opening door
- Ensure mold is level
6. Demold & Cut¶
- Cool to room temperature
- Carefully peel cured PDMS from mold
- Cut to required size with blade
- Storage: Room temperature, long-term stable
Quality Check¶
- Appearance: Transparent, colorless, smooth surface
- Elasticity: Good elasticity, not easily torn
- Bubbles: Minimal in surface and interior
- Thickness: Uniform throughout
Troubleshooting¶
| Problem | Cause | Solution |
|---|---|---|
| Sticky, doesn't set | Inaccurate ratio or poor mixing | Ensure precise weighing and thorough mixing |
| Too many bubbles | Insufficient degassing | Extend vacuum time or degas in stages |
| Uneven thickness | Mold not level or poured too fast | Level mold, pour slowly, gently tilt to spread |
| Incomplete curing | Temperature or time insufficient | Extend curing time or increase temp (max 100°C) |
Notes¶
- Components A and B should be at room temperature before mixing
- Mix in well-ventilated area
- Wear gloves; avoid skin contact with uncured PDMS
- Cured PDMS can be autoclaved (121°C, 20 min)
- Clean cutting tools to avoid contamination
- Clean PDMS surface with ethanol or isopropanol before use
Curing Reference¶
- Room temperature: 24 h
- 80°C: overnight
- 100°C: 2 h
- Standard ratio (10:1): moderate hardness and elasticity
- Higher ratio (12:1): harder, more brittle, better transparency