From 2011.igem.org

Reflectin Thin Films I

The aim of these experiments was to reproduce the flowcoating experiments reported in the literature and pioneer spincoating techniques in addition to performing controls to confirm that any properties of the films made were due to reflectin and not the solvent used in the preparation of the films.

Practice

• The vacuum centrifuged protein was now visible as white pellets on the base of eppendorf tubes.
• This pellet was resuspended in HFIP before being spincoated or flowcoated onto silicon wafers which were prepared as described in the Substrate Preparation Protocol.

Results

Observations

• The first reflectin thin films produced some astounding colours, but were far from uniform.
• Dark objects can be seen in the centre of the regions that vary most in colour which suggests the presence of impurities and diffuse scattering of light. The interference rings suggests the height of the thin film is varying around these impurities.
• As a control we prepared a HFIP (solvent) only thin film which produced a predominantly colourless, dirty smear not a smooth colourful thin film effectively ruling out the involvement of the solvent.

Our first reflectin thin film

One of our first reflectin thin film under the microscope.

HFIP only control

Summary

• The first thin films exhibits thin film interference effects which are not due to the solvent.
• To further investigate the production of more uniform thin films we altered the protocols we were using by namely;
  1. Removal of insoluble impurities in reflectin solution by centrifugation
  2. Reduced spin time from 3000rpm for 1min to 3000rpm for 30s
  3. Use CO₂ pressure wash not O₂ plasma for finer cleaning
  4. A further control with BSA (Bovine Serum Albumin) to test if the effects were unique to reflectin, or a feature of any proteinaceous thin film. (See Reflectin Thin Films II for more details)

Safety

• Safety considerations for the substrate preparation, flowcoating and spincoating techniques can be found on the relevant protocol pages.
• In general labcoat, safety glasses and nitrile safety gloves were worn throughout these three procedures, and all activities took place in the fume hood, except preparation of the silicon substrate.