ThinCert (Ø 24 mm) Centring Holder

thingiverse

Our laboratory uses Greiner ThinCert inserts [6-well, Ø 24 mm, cat # 657638 (1)] as build surfaces for 3D bioprinted tissues created with our Aspect Biosystems RX-1 Bioprinter (2). They work wonderfully, except that the inserts do not always sit central in the plate [Greiner cat # 657160 (3)] and can move around, making long-term live-cell imaging a challenge. These holders were designed to perfectly centre the Thincert in each well of the plate and hold it firmly in place. This greatly improves stability for long-term live-cell imaging, and simplifies our Cytation 5's automatic imaging workflow; we can automatically set bounds within the well for montage imaging without fear of missing one edge of our tissues, and without needing to capture an excessive number images just to make sure we 'get everything'. The holder was also designed such that a Pasteur pipet can be used to aspirate the media without removing the holder or the insert, and media can easily be added with a P1000 or ≤5 mL serological pipet. 1. <a href="https://shop.gbo.com/en/row/products/bioscience/cell-culture-products/thincert-cell-culture-inserts/thincert-cell-culture-inserts-6well/657638.html">Greiner Thincert Inserts, 657638</a> 2. <a href="https://www.aspectbiosystems.com/technology#bioprinter">Aspect Biosystems RX-1 Bioprinter</a> 3. <a href="https://shop.gbo.com/en/row/products/distribution/analyze/5_0060_0020/5_0060_0020_0020/657160.html">Greiner 6-well Plates, 657638</a> *If you use this design in a scientific manuscript, I only ask that you please state the design was received from Dr. Adrian West, cite this webpage, and send me a link to your published paper!* <b>NOTE: I am putting print instructions here, because Thingiverse does not properly parse paragraphs in the print settings section below.</b> This is a very easy print if your printer is well-dialled-in. If you cannot get the holder to fit snug within both the plate and the insert, perhaps just adjust horizontal expansion (±0.2 mm) and/or flow rate (±2%). Also sure your build surface is very level if you are printing large batches, and make sure you use a quality adhesion aid to prevent warping. I would strongly recommend no brim, which would affect how the holder fits into plates and dishes. With the Ultimaker S3 we use a 0.25 mm AA print core and 0.1 mm layer height to ensure utmost surface quality, but print time is approximately one hour per holder. Wider nozzles and taller layer heights will allow you to print much faster. Just make sure your walls/top/bottom are thick enough that the holder prints solid. Material choice depends on how well you can prevent the holder from touching media. In most cases, BASF Ultrafuse PET that has been thoroughly cleaned with 70% EtOH is ideal. For longer term culturing where contamination might be an issue, Ultimaker Polycarbonate that has been thoroughly rinsed, boiled, soaked in 70% EtOH, then UV-exposed would be appropriate.