Leiden Chamber (spaceships) for 18 mm circular slides

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SPACESHIPS - Live Cell Imaging Chambers INTRODUCTION: These are STL files for 3D printing "spaceships" (Leiden or imaging chambers) used for live and fixed cell imaging on inverted microscopes. They fit an 18 mm diameter coverslip, making it simple to culture cells directly on coverslips placed in the wells of a 12-well tissue culture plate. Cells can the be fixed and stained, or transferred live to the chamber, and imaged. The chamber holds ~0.75 ml of media, which is sufficient for imaging cells for over 24 hours. For long term imaging we recommend sealing a second coverslip on the top of the chamber with VALAP (1:1:1 mix of vaseline, lanolin and paraffin) to limit dehydration. These chambers can be easily modified using CAD software to add profusion ports, electrodes, or other features. Design fits an 18 mm round coverslip, and two sizes have been included (to fit microscope stages sized for 35 mm and 42 mm dishes). Eight magnets are required per chamber; four for the baseplate and 4 for the top chamber. It is critical that these magnets be positioned such that the polarity of all magnets on the base plate point in the same direction, and that the magnets on the top-chamber are positioned in the opposite direction (such that they hold the two halves together). To simplify this process, we have included two tools for positioning the magnets in the baseplate and chamber. You will also need an o-ring for each chamber; this is what forms the water-tight seal. The o-ring diameter is critical, but the thickness can vary slightly depending on what is available to you. This chamber has been designed to seal well with #1.5 thickness (0.17 mm) coverslips, but any thickness slip from #1 through #3 should work. FILES: -LCIC_35mm_base.stl - chamber sized to match a 35 mm diameter culture dish -LCIC_42mm_base.stl - chamber sized to match a 42 mm diameter culture dish -MagAlignTools.stl - printable tools to help with magnet alignment -Readme.txt - this file REQUIRED PARTS: Magnet: -8 per chamber, rare-earth magnets (e.g. neodymium) are recommended -cylindrical magnets required: -2.0 to 2.5 mm in diameter -1.5 to 2 mm in height O-Ring: -1 per chamber -17.8 mm outer diameter -1.8-2.5 mm cross-sectional diameter ASSEMBLY: 1) Print the magnet insertion tools and insert a magnet into each tool. Insert the magnets such that they hold the tools together if you touch the tools to to each other. Once you have the magnets inserted properly, hold them in place with a drop of cyanoacrylate (krazy/instant) glue. These tools can be used to assemble multiple printers, so they only need to be made once. 2) Print the base and top chambers. We recommend PLA as this plastic has been previously shown to be cell-safe. Other plastics may work, but check them for cytotoxicity before use. We recommend printing with a 0.4 mm nozzle, 0.2 mm layer thickness, and to print walls at least 5 perimeters thick. Smoothing the final layer is unnecessary. 3) Pickup magnets with the rod-shaped assembly tool, and insert them into the corresponding insets in the base plate. Once you are certain all the magnets are aligned properly, glue them in place with cyanoacrylate glue. 4) Pickup magnets with the tab-shaped assembly tool, and insert them into the corresponding insets in the chamber. If this is done correctly, the chamber should stick by magnetism alone to the base plate. Once you are certain all the magnets are aligned properly, glue them in place with cyanoacrylate glue. 5) Once all glue has cured, insert an o-ring into the groove in the upper chamber, place an 18 mm coverslip into the base plate, and assemble. The two sections should stick to each other and form a water-tight seal. 6) Chambers can be sanitized with an 70% ethanol wash followed by UV treatment. Chambers are reusable, but should be replaced if they start to show signs of distortion, cracking, layer separation, or if cells start to behave unusually.