Slides Microfabrication

This protocol is used for drilling thin coverslips and constructing flowcells based on coverslips.

Contents

1. Drill coverslips using drill press
2. Clean coverslips
3. Construct slides flowcells

1. Drill coverslips using drill press

1. Two coverslips (Fisherbrand, 12-545-M, 24*60-1, LOT #092613-9) will be drilled at the same time. Put two coverslips together and overlap them.
2. Mark two positions where the holes will be drilled on coverslips.
3. Sandwich the two overlapped coverslips between two microscope slides (Fisherbrand, 25*75*1 mm, Cat # 12-552-3).
4. Apply even pressure with two small binder clips. Before drilling, remove steel holders of each clip to ensure the sandwiched slides stable during drilling.
5. Drill the sandwich composition at two marked positions following the ‘’Drill New Slides’’ part in protocol preparing_flowcells_for_single_molecule_experiments.

2. Clean coverslips

1. Following the “Wash slides” part in protocol preparing_flowcells_for_single_molecule_experiments.

3. Construct slides flowcells

1. Cut off two 3*3 mm square parts on double-sided tape. The center distance of two square parts is the distance of the two drilled holes on coverslip.
2. Glue two drilled coverslips by placing the double-sided tape with square parts in the middle.
3. Make a flow chamber following the “Constructing flowcells” part in preparing_flowcells_for_single_molecule_experiments. The two glued coverslips is used as the wafer slide used in normal flow cell construction.

• Note: after sandwich the flow cell between two microscope slides, apply even pressure with four small binder clips and bake in vacuum oven at 120˚C for 30 minutes. 1 hour incubation may easily break coverslips.