Overview¶
Embedding: Photocrosslinking holds a sample in a PEG-nb gel that sets under 405 nm light. The precursor stays liquid until it is exposed, so the sample can be loaded and checked before the gel forms. This page covers Gel: PEG-nb. A digital light processing (DLP) projector exposes the plate at 405 nm for 60 s.
Photograph of photopatterning in a 96-well plate: a digital light processing projector on a stand exposes the plate to 405 nm light.
Precursor, per 1 mL.
| Component | Concentration | Amount |
|---|---|---|
| 4-arm PEG-norbornene | 80 mM | 0.4 g |
| 4-arm PEG-thiol | 20 mM | 0.04 g |
| LAP | 16.9 mM | 4.97 mg |
Materials and Equipment¶
The precursor materials are listed under Materials on Gel: PEG-nb.
Equipment.
| Item | Used for |
|---|---|
| DLP projector (Zwants Supplies Engineering, PRO4500-92-405) | Exposure |
| 96-well or 384-well plate | Plating |
Protocol¶
Prepare the Precursor¶
The precursor is photosensitive
Prepare the precursor in a dark room, and wrap the vial in foil for storage.
Dissolve the components.
Dissolve the PEG-nb, the PEG-thiol and the LAP in 1 mL of the outer solution the liposomes were prepared in.
Mix until clear.
Vortex at 2000 rpm for 1 min, then leave at room temperature for 3 to 5 min.
Vortex again for 2 to 4 min and check that every component has dissolved.
Repeat until the solution is clear.
Mix with the Sample and Plate¶
Mix the sample with the precursor.
Combine 15 µL of sample with 15 µL of precursor.
Plate the mixture.
Dispense 30 µL per well into a 96-well or 384-well plate. Plate each sample in duplicate.
Plate the sample in its outer solution, with no gel, as a reference.
Expose¶
Photopattern the gel.
Expose the plate for exactly 60 s at 405 nm with a DLP projector.
The precursor can be diluted in the well
When a lower concentration is needed, dilute the precursor 1:1 in the well, for example 50 µL of precursor with 50 µL of buffer.
Supported by¶
DevNotes that support this page include:
