Bath Imaging Chamber
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A chamber to hold cells and tissue
Herein is a design for holding and bathing of cell culture and tissues during electrophysiological recording/stimulation and/or optical imaging, and can even be used in conjunction with patch clamp (dare we say Dynamic Clamp???). The chamber offers several methods to connect various tube sizes to introduce fluid exchange at (or near) laminar flow. It is compatible with a wide variety of microscopes and emphasizes room for larger objectives. The chamber sits on top of a basic coverslip and can be held down with magnets to a stage adapter. Thin tissue can be held in place with clear fishing line, strung between two additional parts.
Check out another project from the ONE Core, a Temperature Controlled Slice Holder.
Parts
All designs shown here can be 3D printed. The 3D printed parts should be printed in a high resolution, biosafe material such as SLA MED610. Use a single #1 coverslip (thickness around 0.15 mm or thereabouts). Magnets can be glued in place and should be 1/16” round by 1/16” mm high, such as the K&J D11-N52.
The designs are named (and color coded in the image above): Tissue Top (green), Tissue Bottom (orange), Chamber (blue); Hold Fast (grey) (print two!), and Stage (yellow). Not shown: magnets, tissue, coverslip, tubes, and imaging/recording/stimulation media.
| Name | File Type | Design/Description |
|---|---|---|
| ElectroPhysiologyChamberv16Chamber.ipt | ipt | Chamber |
| ElectroPhysiologyChamberv16Chamber.stl | stl | Chamber |
| ElectroPhysiologyChamberv16SideHoldfast.ipt | ipt | Hold Fast |
| ElectroPhysiologyChamberv16SideHoldfast.stl | stl | Hold Fast |
| ElectroPhysiologyChamberv16Stage.ipt | ipt | Stage |
| ElectroPhysiologyChamberv16Stage.stl | stl | Stage |
| ElectroPhysiologyChamberv16TissueBottom.ipt | ipt | Tissue Bottom |
| ElectroPhysiologyChamberv16TissueBottom.stl | stl | Tissue Bottom |
| ElectroPhysiologyChamberv16TissueTop.ipt | ipt | Tissue Top |
| ElectroPhysiologyChamberv16TissueTop.stl | stl | Tissue Top |
Assembly of the Tissue Holder
This chamber can be used without any tissue holder, but can also hold tissue sandwiched between overlapping lines of clear fishing line. Here’s how to assemble these parts:
- Take the fishing line and tie it to the side post, or the shaft/peg shown in green.
- Feed the line in the direction of the red arrows. That is, across to the other side (perfectly perpendicular to the sides), around the ‘peg’ on the opposite side and back. Repeat back and forth until complete.
- Tie the end onto the side post or the end as shown in yellow. Do not overtighten the line, as this can at best snap the line and at worst damage the pegs or part as a whole. How tight you make this will effect how much slack there is in the line, and therefore the gap between the fishing line ‘sandwiches’: The thickness allotted to the tissue.
- Do this for the Tissue Bottom as well.
- You can then load the tissue in solution onto the top surface of the fishing line ‘sandwich bottom’ of the Tissue Bottom part (#2 in the picture below), create the ‘sandwich’ by placing the Tissue Top on top of the tissue (#1), and placing the Tissue Top and Bottom (now a sandwich) into the chamber (#3). When put together (1 above 2, and 2 above 3), your assembly should look like #4 below.
We have heard rumor that the tissue can float to the top. We are surprised by this, but wish to squash any further rumors by designing in holes that can hold the same magnets as used to hold the chamber to the stage (1/16” round, 1/16” thick). They can be glued in place where indicated by the blue arrows in the picture above.
Assembly of the Rest
Up to twenty magnets can be glued into the Stage and the corresponding Hold Fasts. Then 1/4” tubing can be affixed into the fluid entry (the end of the Chamber with the ‘bridge’ to stop bubbles) and the fluid exit (the Chamber side that would not block bubbles). You can use the T-Barb connectors or just use twist-ties to attach the tubing onto the half tubes.
ONE Core acknowledgment
Please acknowledge the ONE Core facility in your publications. An appropriate wording would be:
“The Optogenetics and Neural Engineering (ONE) Core at the University of Colorado School of Medicine provided engineering support for this research. The ONE Core is part of the NeuroTechnology Center, funded in part by the School of Medicine and by the National Institute of Neurological Disorders and Stroke of the National Institutes of Health under award number P30NS048154.”
