At our company we might be interested in buying a liquid handler to process cells. The process is as follows:
- Removing supernatant without disturbing a cell pellet
- Resuspending in a fixed volume.
- Counting the amount of cells
- Dilute to a certain amount of cells/mL
- Transfer to X number of tubes depending on volume.
We might be interested in buying a Tecan Fluent for that, but does anyone have experience with cell handling and cell counting with the Tecan Fluent?
And if anyone has experience… what are the pro and cons on this system with this ?
A friend of mine is also assessing a project similar to how you describe, and was looking into utilizing this on their current Fluent system: Live cell imaging. I’m not sure of the outcome as it’s planned for 2024/2025, but I can follow up if I hear anything.
So long as your labware is uniform, I don’t see why you can’t perform steps 1 and 2 on the main Fluent worktable (deck), count the cells in step 3 with the Spark above, move the sample back onto the worktable and then perform 4 and 5.
@Shaik Likely has some recommendations for this.
I mean it’s a general purpose automated liquid handler with a variety of tools that come standard or can be integrated into the system. It can handle the tasks.
Question for you: how are you cell counting? Is the Tecan the driving force or is this being integrated into a scheduler? In addition, what’s your starter labware, intermediate and final labware of choice?
At the moment the start labware is 15mL tubes in a cooled Rack (hope to get that to 5mL ones in cooled Rack), then remove buffer and resuspension with new buffer in same tube. After mixing we will take a sample and transfer to 96-flat-bottom for measurement. After measurement we use the data for an in situ normalization and then transfer to multiple 2mL screw-cap tubes.
But to be honest, most of this labware is what we have around the labs at the moment. Could change if something better is available.
cell counting is done with:
- countess or
- Cellaca Cell Counter
As for the cell counting: we might consider the use of the Tecan Spark Cyto.
Does anyone has experience with that equipment and the integration in a Fluent 480?
Additionally, I’ve heard that for the use of flat-bottom plates a tilting device is sometimes required to make sure all the volume is removed from inside the well. Does anyone have know if this would ‘really’ make a difference. I’ve tried to remove all the liquid from a 96-well flat bottom and then the residual volume is ~5uL. Will the tilting device help to remove it all?
I’ve used the Spark but not the Spark Cyto.
It does help to remove as much of the media as you can. A tilt module will help but also the micro script is powerful enough so that you could… in theory, create a circular pipetting motion which may offset the need for tilting module.
It might take a little longer but save funds in the long run.
A tilt module will help
Just chiming in here:
@dirkvanessen From personal (painful) experience, I would highly recommend against purchasing Tecan’s tilt module. Not only is it a huge pain to program, it is over-engineered and the degrees of tilt will skew overtime. If you decide to go the tilting route, use a third party one
Thanks, that is good to know. Do you have experience with third party tilt modules and how to integrate into a Tecan Fluent?
I have never actually gotten around to using another party’s tilt module. My solution was… a bit ridiculous actually
Link to gif here in case it doesn’t work
I used a 3d printed tilter that we knew the exact degrees of, and used their tilter to transfer my plate to this more reliable one
The problem with their tilter is it has 180 degrees of rotation, which is more than you really need for cell culture, where I just needed either TILTED or NOT TILTED. To make it more fun, when specifying a degree, the degree-offset of this device can change overtime if you crash into it by accident while testing, so it’s not quite reliable in positioning.
So I just used it to transfer my plate to a 3d printed static tilter
I know this doesn’t really help you, but it’s a fun story. For integration of a 3rd party tilter, I think you can pay Tecan to write you a driver. I have no idea how much that costs
i recently completed a tilting module for integration onto Tecan EVO or Fluent,
please reach out for more details
I dig this ridiculous solution! That’s excellent.
This is awesome! In a similar vein, I believe people have 3D printed tilt modules that use a channel to physically move to a position and tilt a plate by hitting some corner of the 3D printed carrier.
I believe @cyancey may have once shared a video of it on an Opentrons on LinkedIn.
Yes! I saw a webinar from opentrons from Monomer Bio that had a 3d printed tilt module that was basically a see-saw that you could hit with a pipette channel! It was so cool and ingenious!
I contacted the individual who designed it, and they said they’d send me the CAD files designs, but that was a long time ago. Maybe I should poke them again
EDIT: Found it here!
Are you currently using the Cellaca? Thoughts?
FYI @Shinedalgarno @dirkvanessen
QInstruments just announced their tilting module called the TiltStation.
Same footprint as their shakers but the tilts seems fixed. You can control the speed at which it tilts and make it seesaw. Worth checking out at SLAS.
What’s your throughput goals here? A Lynx VVP would do this much faster.