[CNC] [Request] Designing/cutting Lab equipment for the real time visualization of DNA strand exchange

[Lukas Lindorfer]

Hi Armin,

my Outlook put you into „Spam“, sorry about that. The cell sould be made of aluminum. The channels are, as discussed before, 4mm wide to fit FPLC screws which can in turn lead a thin tube to the cell. A threading would be needed, I’ll copy what I wrote Sebastian:

https://www.fishersci.co.uk/shop/products/peek-finger-tight-connector/11390052

We would drill a M5 thread, pitch of 0.8 into the holes to lead a very thin tube (1/16 inch) through them. Due to this tubing, the smoothness of the hole is not really a factor.


Thanks a lot for your interest, guys! You can reach me anytime at Lukas.lindorfer at hotmail.com<mailto:Lukas.lindorfer at hotmail.com> if you have further questions.

Best, Lukas


________________________________
Von: CNC <cnc-bounces at lists.metalab.at> im Auftrag von Armin Faltl <armin.faltl at aon.at>
Gesendet: Monday, October 28, 2019 2:27:12 PM
An: cnc at lists.metalab.at <cnc at lists.metalab.at>
Betreff: Re: [CNC] [Request] Designing/cutting Lab equipment for the real time visualization of DNA strand exchange

Hi!

sounds interessting to me too.

What material shall the cell be made of?
Regarding the angles, I think we can make a fixture.
IIRC, I do have drills down to 0.4mm diameter.
What type of connections do you need at the exits of the cell?

Best Regards, Armin


On 28.10.19 14:12, Sebastian Bachmann wrote:
> Hi!
>
> That sounds interesting!
> I'm just now looking at some pictures in the internetz of those flow cells
> and it looks like the cavity is in the micrometer range - usally like 400µm.
> When looking at the screenshots, I suspect that the dimensions are in mm, hence
> the hole should be 500µm in diameter - right?
> So I would say, this should be do-able in the CNC using regular drills.
> An important question is the required surface structure of the holes.
> I would guess, that the surface must be very smooth, in order to not damage
> cells and also provide a laminar flow through the cell?
> This might be problematic when using regular drills, as they do not leave a nice
> surface finish.
> You can get 0.5mm reamers but they are horrible expensive: https://www.accu.co.uk/en/machine-reamers/444489-221450-0500?PageSpeed=noscript
> If you break one of these ... also our CNC can not run with low speeds, but
> maybe at this size the lowest rpm of 6000 might be sufficient.
>
> So in theory, you could build this thing using a three step process:
> 1) mill the pockets on the lower surface
> 2) mill the first wedge and drill holes
> 3) mill the second wedge and drill holes
>
> For steps 2 and 3 you need either some angle plate or a sine table - we dont
> have either of those in the lab.
> There might be working drills in the lab, I think they have 0.75mm diameter -
> not sure how critical the dimensions are.
> Drills are cheap - everything else is not. So depending on the surface, you can
> get it cheap or horrible expensive.
>
> Also note, that our CNC is right now in some undefined state where our spindle
> drive has some undefined issues.
>
> Sebastian
>
>
>
> On Mon, Oct 28, 2019 at 12:00:47PM +0000, Lukas Lindorfer wrote:
>> Hi all,
>>
>> my name is Lukas.
>> The Metalab was recommended to me by my friend Juli („Juli wie August“) as a place to develop and share ideas. Right now I am sitting in your main room for the first time, typing up an idea that might interest some of you. I am a biology student currently working in a group doing research on on the generation and analysis of proteins involved in plant meiosis. We know what (some of them) do, but we have never seen it “live“ – And I have an idea to change that.
>>
>> I am seeking help in designing  and cutting a flow cell for TIRF microscopy.
>> If anyone is interested in collaborating on this idea with me, I would be very grateful and happy.
>>
>> Attached you will find a screenshot of the prototype. If anyone could spare some time for advice in cutting and design and is interested in the project as well as a beer/dinner/whatever on me, please contact me!
>>
>> Cheers, Lukas
>>
>>
>> More Infos:
>>
>> The „state of the art“ is applying the proteins to investigate at the lab bench and then going to the laser microscope. The Problem: Those proteins work fast – less than five minutes. When I arrive at the microscope, the reaction is already finished which makes visualizing/filming it rather hard considering sample preparation takes up to 3 hours.
>>
>> This is where the flow cell comes in. With this system, we could prepare the protein sample in the microscope itself – visualizing meiotic strand exchange in real time for the first time in history!
>>
>>
>> Some vocabulary:
>>
>> Meiotic Strand Exchange is the exchange of DNA between two chromosomes in the generation of sperms/eggs --> The stuff that makes every human unique!
>> https://www.yourgenome.org/facts/what-is-meiosis
>>
>> TIRF (Total Internal Reflection Fluorescence Microscopy) is a niche laser microscopy technique, being able to visualize single molecules linked to a glass plate. https://www.microscopyu.com/techniques/fluorescence/total-internal-reflection-fluorescence-tirf-microscopy)
>>
>> A flow cell is exactly what it sounds like: A cell stuff flows through. In our case, the glass plate to investigate would be rubber cemented on the bottom of the cell so it can be looked at with the laser.
>>
>>
>
>
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