From postmaster at grafoo.at Sat Oct 5 14:36:10 2019 From: postmaster at grafoo.at (Manuel Graf) Date: Sat, 5 Oct 2019 14:36:10 +0200 Subject: [CNC] introduction workshop Message-ID: is there any introduction workshop planned for the near future? if so I'd glad to take part. cheers, grafoo -------------- next part -------------- An HTML attachment was scrubbed... URL: From peter at null.priv.at Sat Oct 5 14:48:42 2019 From: peter at null.priv.at (Peter Kuhm) Date: Sat, 5 Oct 2019 14:48:42 +0200 Subject: [CNC] introduction workshop In-Reply-To: References: Message-ID: <20191005144842.1101db82@zero> On Sat, 5 Oct 2019 14:36:10 +0200 Manuel Graf wrote: > is there any introduction workshop planned for the near future? if so I'd > glad to take part. > cheers, grafoo metoo :) From metalab at reox.at Thu Oct 24 08:57:21 2019 From: metalab at reox.at (reox) Date: Thu, 24 Oct 2019 08:57:21 +0200 Subject: [CNC] =?utf-8?q?VFD-Probleme_bei_der_CNC-Fr=C3=A4se?= In-Reply-To: <20190819153607.GB1882@helios> References: <75efe98b-7627-49c6-c52d-c7d32bedb1ec@lightbug.at> <20190812184206.GG1881@helios> <5060fdfd-5a64-f303-e88d-dceaa2e9c8c7@wireloss.net> <20190819153607.GB1882@helios> Message-ID: Ahoi! Treten die Probleme mit der CNC jetzt immer noch auf? Falls Ja, mag sich jemand mit mir dem VFD n?chste Woche annehmen? LG Sebastian On 2019-08-19 17:36, Sebastian Bachmann wrote: > Hi! > > On Wed, Aug 14, 2019 at 04:50:41PM +0200, Clemens Hopfer wrote: >> Hi, >> >> Am 12.08.19 um 20:42 schrieb Sebastian Bachmann: >> > 2) Herr Kloss meint, dass kaputte Elkos diese Unterspannungsfehlermeldung >> > ausl?sen. Er meint auch, es lohnt da kein Tausch der Elkos sondern eher gleich >> > einen gescheiten VFD kaufen - er hat auch nur noch Omron im Angebot. >> >> das klingt f?r mich realistisch, je nach Qualit?t trockenen die nach >> einiger Zeit aus. > > Ja, so vom Gef?hl her w?re das auch immer das erste woran ich denke > w?rde. > >> >> Ich hab ein Fluke Scopemeter, mit dem ich schon einen defekten 2,2kVA >> FU >> im Funkfeuer Housing durchgemessen hab, der dort 4 St?ck 60cm >> Industriel?fter antreibt. >> >> Ich kann mir das nach dem Camp mit jemandem, der sich mit der CNC >> auskennt mal anschauen. > > Ok cool! Ich k?nnte mir das wohl mit dir ansehen aber bin leider etwas > verplant. > Das n?chste freie Wochenende ist bei mir erst der 21.09.... > Unter der Woche w?rde es vllt mal am Abend gehen. > >> Bei dem FU den ich damals debugged habe war eine der >> Stromsteller-Stufen >> einer Phase kaputt, damit konnten die Motoren zwar anlaufen aber >> hatten >> kaum Leistung und der FU ging in ?berlast. Ob die Kondensatoren hin >> sind >> sollte man gut sehen. Wenn der FU nicht vergossen ist, ists wohl recht >> leicht die zu tauschen. > > Soweit ich gesehen habe, kommt man an alle Boards einfach heran. > >> >> > 3) Wir haben keinen Netzfilter vor dem VFD. Er empfiehlt jedenfalls den Einsatz >> > eines solchen. Evt kommen die Probleme auch daher, da hab ich aber keinen Plan >> > wie man das testen kann. >> >> Das klingt f?r mich unrealistisch, den Netzfilter will man normal >> haben, >> um nicht im Netz St?rungen zu verursachen, den FU juckt das normal >> wenig. > > Mhh, da m?sste ich den Herrn Kloss nochmal fragen was seine > Argumentation ist. > Evt k?nnten die St?rungen die der FU verursacht das Breakoutboard aus > dem Takt > bringen, was sich dann zum FU auswirkt? > Der G4 hat einen Netzfilter eingebaut, vgl > https://metalab.at/wiki/images/a/a2/Icnc.svg > > >> >> > 4) Als Ersatz gibt es diesen hier: https://www.sorotec.de/shop/Omron-FU-0-75KW-200V-1-Phasig.html >> > und diesen Filter: https://www.sorotec.de/shop/Netzfilter-1-Phasig-f-r-FU-0-75KW.html >> >> > wenn wir ganz lustig sind, kaufen wir einen Bremswiderstand auch mit ;) >> > https://www.sorotec.de/shop/Bremswiderstand-70Ohm-0-2kW.html >> >> Kenne ich nur f?r gr??ere Massen, die gebremst werden m?ssen. > > Damit kann man die Spindel sehr viel schneller abbremsen. Aktuell l?uft > sie > einfach aus. > > LG Sebastian > >> >> Lg, >> Clemens >> >> > On Mon, Aug 12, 2019 at 08:21:51PM +0200, ripper wrote: >> >> Servus, >> >> >> >> Das, bereits vor einiger Zeit angesprochene, Problem mit dem VFD (Siehe >> >> Thread aus dem April 18) besteht weiterhin. In unregelm??igen Abst?nden >> >> f?llt w?hrend dem Fr?sen die Drehzahl ab, teilweise bis zum absoluten >> >> Stillstand der Spindel. Der VFD meldet daraufhin "E.LU" was laut [1] f?r >> >> ":lower voltage in DC bus" steht, was auch die Diagnose vom letzten Mal >> >> war. Die CNC-Fr?se wurde daraufhin als repariert gemeldet, da die >> >> Verkabelung in Ordnung sei. >> > >> > Vor allem konnte der Fehler nicht mehr reproduziert werden. >> > Wo ich angefangen hatte das zu debuggen war er st?ndig da und auf einmal lie? er >> > sich nicht mehr provizieren. >> > K?nnte eine thermische Komponente haben. >> > >> >> >> >> Meine Beobachtungen dabei: Wenn die Drehzahl (der Ger?uschentwicklung >> >> nach) abf?llt und die Ausf?hrung pausiert wird, "erholt" sich der VFD >> >> wieder. Wirkt auf mich nach einem Kabelbruch der Motorleitung in der >> >> Schleppkette, auch wenn die Fehlermeldung nicht dazupasst? >> > >> > Beim letzten Mal waren die Probleme nur in bestimmten Drehzahlbereichen - >> > korrekt? Das w?rde mMn gegen einen Kabelbruch sprechen. >> > Andererseits, der Kabelbruch k?nnte wie ein Kondensator wirken und bei >> > bestimmten Drehzahlen wilde Dinge tun - damit w?re das Parallelkabel zwischen PC >> > und Breakoutboard im Verdacht, da nur hier Pulssignale ?bertragen werden. >> > >> > Was mir auch noch einf?llt: Die beiden Relais auf dem Breakoutboard hatten >> > glaube ich mal ein Problem und haben nicht ganz korrekt durchgeschalten. Die >> > sollte man sich evt auch nochmal genauer ansehen. >> > >> > LG Sebastian >> > >> > >> >> >> >> LG Michael >> >> >> >> [1] http://alis.kms-bg.com/catalogue/PI9000_operation.pdf, Seite 45 >> >> >> >> -- >> >> ________________________________________ >> >> https://metalab.at/wiki/Benutzer:ripper >> >> >> >> >> >> >> >> _______________________________________________ >> >> CNC mailing list >> >> CNC at lists.metalab.at >> >> https://lists.metalab.at/mailman/listinfo/cnc >> > >> > _______________________________________________ >> > CNC mailing list >> > CNC at lists.metalab.at >> > https://lists.metalab.at/mailman/listinfo/cnc >> > >> > > > > >> _______________________________________________ >> CNC mailing list >> CNC at lists.metalab.at >> https://lists.metalab.at/mailman/listinfo/cnc > > > _______________________________________________ > CNC mailing list > CNC at lists.metalab.at > https://lists.metalab.at/mailman/listinfo/cnc From postmaster at grafoo.at Thu Oct 24 09:28:13 2019 From: postmaster at grafoo.at (Manuel Graf) Date: Thu, 24 Oct 2019 09:28:13 +0200 Subject: [CNC] einschulung Message-ID: hallo, hat demn?chst mal jemand f?r einen einf?hrungsworkshop zeit? nachdem ich schon herumgefragt habe und auch andere interesse haben, w?hren wahrscheinlich um die 4 teilnehmer dabei. -------------- next part -------------- An HTML attachment was scrubbed... URL: From metalab at reox.at Thu Oct 24 09:38:29 2019 From: metalab at reox.at (reox) Date: Thu, 24 Oct 2019 09:38:29 +0200 Subject: [CNC] einschulung In-Reply-To: References: Message-ID: Hi! Nachdem die CNC in einem undefinierbaren-keiner-wei?-ob-der-VFD-nicht-doch-kaputt-ist Zustand befindet, kann man nicht so wirklich einen Workshop machen. Ungeachtet dessen sind wie immer folgende Fragen zu kl?ren: * Was soll gefr??t werden? * gibt es G-Code? * wenn nein, kann man den erzeugen? * Sind werkzeuge wie fr?ser etc vorhanden? * Wie soll die Aufspannung vom Werkst?ck funktionieren? Das kann man sich sicher mal vorher anschauen, dazu braucht man die Fr?se nicht mal einschalten. Wenn ihr ein Projekt habt, welches man gleich als Anschuungsobjekt verwenden kann, dann kann man das gleich beim Workshop besprechen. LG Sebastian On 2019-10-24 09:28, Manuel Graf wrote: > hallo, > hat demn?chst mal jemand f?r einen einf?hrungsworkshop zeit? > nachdem ich schon herumgefragt habe und auch andere interesse haben, > w?hren wahrscheinlich um die 4 teilnehmer dabei. > _______________________________________________ > CNC mailing list > CNC at lists.metalab.at > https://lists.metalab.at/mailman/listinfo/cnc From harald.boeck at chello.at Thu Oct 24 22:25:29 2019 From: harald.boeck at chello.at (Harald) Date: Thu, 24 Oct 2019 22:25:29 +0200 Subject: [CNC] einschulung In-Reply-To: References: Message-ID: <000001d58aa9$2f0042b0$8d00c810$@chello.at> Hallo die CNC-Fr?ser, ich w?re an einem Workshop interessiert, kurze theoretische Einf?hrung inklusive. Kann auch gerne bei der FVD fact finding mission helfen. LG Harald -----Urspr?ngliche Nachricht----- Von: CNC Im Auftrag von reox Gesendet: Donnerstag, 24. Oktober 2019 09:38 An: Neigungsgruppe Zerspanungstechnik/CNC und Werkstatt allgemein Betreff: Re: [CNC] einschulung Hi! Nachdem die CNC in einem undefinierbaren-keiner-wei?-ob-der-VFD-nicht-doch-kaputt-ist Zustand befindet, kann man nicht so wirklich einen Workshop machen. Ungeachtet dessen sind wie immer folgende Fragen zu kl?ren: * Was soll gefr??t werden? * gibt es G-Code? * wenn nein, kann man den erzeugen? * Sind werkzeuge wie fr?ser etc vorhanden? * Wie soll die Aufspannung vom Werkst?ck funktionieren? Das kann man sich sicher mal vorher anschauen, dazu braucht man die Fr?se nicht mal einschalten. Wenn ihr ein Projekt habt, welches man gleich als Anschuungsobjekt verwenden kann, dann kann man das gleich beim Workshop besprechen. LG Sebastian On 2019-10-24 09:28, Manuel Graf wrote: > hallo, > hat demn?chst mal jemand f?r einen einf?hrungsworkshop zeit? > nachdem ich schon herumgefragt habe und auch andere interesse haben, > w?hren wahrscheinlich um die 4 teilnehmer dabei. > _______________________________________________ > CNC mailing list > CNC at lists.metalab.at > https://lists.metalab.at/mailman/listinfo/cnc _______________________________________________ CNC mailing list CNC at lists.metalab.at https://lists.metalab.at/mailman/listinfo/cnc From lukas.lindorfer at hotmail.com Mon Oct 28 13:00:47 2019 From: lukas.lindorfer at hotmail.com (Lukas Lindorfer) Date: Mon, 28 Oct 2019 12:00:47 +0000 Subject: [CNC] [Request] Designing/cutting Lab equipment for the real time visualization of DNA strand exchange Message-ID: 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. -------------- next part -------------- An HTML attachment was scrubbed... URL: -------------- next part -------------- A non-text attachment was scrubbed... Name: FlowCell 1.jpeg Type: image/jpeg Size: 31235 bytes Desc: FlowCell 1.jpeg URL: -------------- next part -------------- A non-text attachment was scrubbed... Name: FlowCell 2.jpeg Type: image/jpeg Size: 41764 bytes Desc: FlowCell 2.jpeg URL: From metalab at reox.at Mon Oct 28 14:12:20 2019 From: metalab at reox.at (Sebastian Bachmann) Date: Mon, 28 Oct 2019 14:12:20 +0100 Subject: [CNC] [Request] Designing/cutting Lab equipment for the real time visualization of DNA strand exchange In-Reply-To: References: Message-ID: <20191028131219.fsg4unegdxav4qgk@helios> 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 400?m. When looking at the screenshots, I suspect that the dimensions are in mm, hence the hole should be 500?m 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. > > > _______________________________________________ > CNC mailing list > CNC at lists.metalab.at > https://lists.metalab.at/mailman/listinfo/cnc From lukas.lindorfer at hotmail.com Mon Oct 28 14:26:03 2019 From: lukas.lindorfer at hotmail.com (Lukas Lindorfer) Date: Mon, 28 Oct 2019 13:26:03 +0000 Subject: [CNC] [Request] Designing/cutting Lab equipment for the real time visualization of DNA strand exchange In-Reply-To: <20191028131219.fsg4unegdxav4qgk@helios> References: , <20191028131219.fsg4unegdxav4qgk@helios> Message-ID: Hi Sebastian, thank you for your interest! The difference between the flow cells commonly used and the ones you are probably seeing Right now is that we do not work with whole cells but only with DNA strands and proteins in solution, so a little bit of shearing would not be too terrible. The holes are 4mm wide since they are optimized for those FPLC screws to make a wider range of applications possible: 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. What is critical is to have a really flat ground surface since TIRF microscopy is only practical in a range of about 200nm. I assume (but actually do not know) that the block to cut the cell out of would be rather flat to begin with. Is this the case? The model I sent still has some (a lot of) room for improvement ? Would you be interested in meeting at some point in the next weeks to talk about the cell? That way, I could give you some insights on what we know and need to know. Best, Lukas ________________________________ Von: CNC im Auftrag von Sebastian Bachmann Gesendet: Monday, October 28, 2019 2:12:20 PM An: Neigungsgruppe Zerspanungstechnik/CNC und Werkstatt allgemein Betreff: Re: [CNC] [Request] Designing/cutting Lab equipment for the real time visualization of DNA strand exchange 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 400?m. When looking at the screenshots, I suspect that the dimensions are in mm, hence the hole should be 500?m 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. > > > _______________________________________________ > CNC mailing list > CNC at lists.metalab.at > https://lists.metalab.at/mailman/listinfo/cnc _______________________________________________ CNC mailing list CNC at lists.metalab.at https://lists.metalab.at/mailman/listinfo/cnc -------------- next part -------------- An HTML attachment was scrubbed... URL: From armin.faltl at aon.at Mon Oct 28 14:27:12 2019 From: armin.faltl at aon.at (Armin Faltl) Date: Mon, 28 Oct 2019 14:27:12 +0100 Subject: [CNC] [Request] Designing/cutting Lab equipment for the real time visualization of DNA strand exchange In-Reply-To: <20191028131219.fsg4unegdxav4qgk@helios> References: <20191028131219.fsg4unegdxav4qgk@helios> Message-ID: 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 400?m. > When looking at the screenshots, I suspect that the dimensions are in mm, hence > the hole should be 500?m 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. >> >> > > >> _______________________________________________ >> CNC mailing list >> CNC at lists.metalab.at >> https://lists.metalab.at/mailman/listinfo/cnc > > _______________________________________________ > CNC mailing list > CNC at lists.metalab.at > https://lists.metalab.at/mailman/listinfo/cnc From metalab at reox.at Mon Oct 28 14:35:36 2019 From: metalab at reox.at (Sebastian Bachmann) Date: Mon, 28 Oct 2019 14:35:36 +0100 Subject: [CNC] [Request] Designing/cutting Lab equipment for the real time visualization of DNA strand exchange In-Reply-To: References: <20191028131219.fsg4unegdxav4qgk@helios> Message-ID: <20191028133536.agowdscddqqmymfa@helios> Hi! On Mon, Oct 28, 2019 at 01:26:03PM +0000, Lukas Lindorfer wrote: > Hi Sebastian, > > thank you for your interest! The difference between the flow cells commonly used and the ones you are probably seeing Right now is that we do not work with whole cells but only with DNA strands and proteins in solution, so a little bit of shearing would not be too terrible. The holes are 4mm wide since they are optimized for those FPLC screws to make a wider range of applications possible: https://www.fishersci.co.uk/shop/products/peek-finger-tight-connector/11390052 Ohh okay, 4mm is not a problem and you can use plenty of tools to get a very smooth finish even! > 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. > > What is critical is to have a really flat ground surface since TIRF microscopy is only practical in a range of about 200nm. I assume (but actually do not know) that the block to cut the cell out of would be rather flat to begin with. Is this the case? No, you can not assume that the blocks are flat. I would assume that you use aluminium? In that case, the flatness is usually in the range of hundrets of ?m. Milling the block flat, should reduce it - ideally you would surface grind it, or lapp the block using a master plate (which we dont have). What is the approximate surface flatness you require? (see also here http://www.engineeringessentials.com/gdt/flatness/flatness.htm for a definition of this measure) You can produce very flat surfaces by lapping three plates against each other. Oxtools has some videos on that: https://www.youtube.com/watch?v=rHmsQEAx16o > > The model I sent still has some (a lot of) room for improvement ? Would you be interested in meeting at some point in the next weeks to talk about the cell? That way, I could give you some insights on what we know and need to know. Next week is very bad for me... I have time this week, and also wanted to visit metalab and the CNC and see whats wrong with the VFD. Sebastian > > Best, > Lukas > > > > > ________________________________ > Von: CNC im Auftrag von Sebastian Bachmann > Gesendet: Monday, October 28, 2019 2:12:20 PM > An: Neigungsgruppe Zerspanungstechnik/CNC und Werkstatt allgemein > Betreff: Re: [CNC] [Request] Designing/cutting Lab equipment for the real time visualization of DNA strand exchange > > 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 400?m. > When looking at the screenshots, I suspect that the dimensions are in mm, hence > the hole should be 500?m 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. > > > > > > > > > _______________________________________________ > > CNC mailing list > > CNC at lists.metalab.at > > https://lists.metalab.at/mailman/listinfo/cnc > > > _______________________________________________ > CNC mailing list > CNC at lists.metalab.at > https://lists.metalab.at/mailman/listinfo/cnc > _______________________________________________ > CNC mailing list > CNC at lists.metalab.at > https://lists.metalab.at/mailman/listinfo/cnc From metalab at reox.at Mon Oct 28 14:38:11 2019 From: metalab at reox.at (Sebastian Bachmann) Date: Mon, 28 Oct 2019 14:38:11 +0100 Subject: [CNC] [Request] Designing/cutting Lab equipment for the real time visualization of DNA strand exchange In-Reply-To: <20191028133536.agowdscddqqmymfa@helios> References: <20191028131219.fsg4unegdxav4qgk@helios> <20191028133536.agowdscddqqmymfa@helios> Message-ID: <20191028133811.vjjmtssgtpdo2zxg@helios> On Mon, Oct 28, 2019 at 02:35:36PM +0100, Sebastian Bachmann wrote: > Hi! > > On Mon, Oct 28, 2019 at 01:26:03PM +0000, Lukas Lindorfer wrote: > > Hi Sebastian, > > > > thank you for your interest! The difference between the flow cells commonly used and the ones you are probably seeing Right now is that we do not work with whole cells but only with DNA strands and proteins in solution, so a little bit of shearing would not be too terrible. The holes are 4mm wide since they are optimized for those FPLC screws to make a wider range of applications possible: https://www.fishersci.co.uk/shop/products/peek-finger-tight-connector/11390052 > > Ohh okay, 4mm is not a problem and you can use plenty of tools to get a very > smooth finish even! > > > 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. > > > > What is critical is to have a really flat ground surface since TIRF microscopy is only practical in a range of about 200nm. I assume (but actually do not know) that the block to cut the cell out of would be rather flat to begin with. Is this the case? > > No, you can not assume that the blocks are flat. I would assume that you use > aluminium? In that case, the flatness is usually in the range of hundrets of ?m. Or less, obviously! There might be suppliers which sell precision stuff - but usually that is steel, which we can not handle in the CNC. > Milling the block flat, should reduce it - ideally you would surface grind it, > or lapp the block using a master plate (which we dont have). > What is the approximate surface flatness you require? > (see also here http://www.engineeringessentials.com/gdt/flatness/flatness.htm > for a definition of this measure) > > You can produce very flat surfaces by lapping three plates against each other. > Oxtools has some videos on that: https://www.youtube.com/watch?v=rHmsQEAx16o > > > > > > The model I sent still has some (a lot of) room for improvement ? Would you be interested in meeting at some point in the next weeks to talk about the cell? That way, I could give you some insights on what we know and need to know. > > Next week is very bad for me... I have time this week, and also wanted to visit > metalab and the CNC and see whats wrong with the VFD. > > Sebastian > > > > > Best, > > Lukas > > > > > > > > > > ________________________________ > > Von: CNC im Auftrag von Sebastian Bachmann > > Gesendet: Monday, October 28, 2019 2:12:20 PM > > An: Neigungsgruppe Zerspanungstechnik/CNC und Werkstatt allgemein > > Betreff: Re: [CNC] [Request] Designing/cutting Lab equipment for the real time visualization of DNA strand exchange > > > > 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 400?m. > > When looking at the screenshots, I suspect that the dimensions are in mm, hence > > the hole should be 500?m 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. > > > > > > > > > > > > > > > _______________________________________________ > > > CNC mailing list > > > CNC at lists.metalab.at > > > https://lists.metalab.at/mailman/listinfo/cnc > > > > > > _______________________________________________ > > CNC mailing list > > CNC at lists.metalab.at > > https://lists.metalab.at/mailman/listinfo/cnc > > > _______________________________________________ > > CNC mailing list > > CNC at lists.metalab.at > > https://lists.metalab.at/mailman/listinfo/cnc > > > _______________________________________________ > CNC mailing list > CNC at lists.metalab.at > https://lists.metalab.at/mailman/listinfo/cnc From lukas.lindorfer at hotmail.com Mon Oct 28 15:03:35 2019 From: lukas.lindorfer at hotmail.com (Lukas Lindorfer) Date: Mon, 28 Oct 2019 14:03:35 +0000 Subject: [CNC] [Request] Designing/cutting Lab equipment for the real time visualization of DNA strand exchange In-Reply-To: <20191028133811.vjjmtssgtpdo2zxg@helios> References: <20191028131219.fsg4unegdxav4qgk@helios> <20191028133536.agowdscddqqmymfa@helios>, <20191028133811.vjjmtssgtpdo2zxg@helios> Message-ID: Hi, I need the cell by January only ? Ideally, we could meet between the 4th of November and the beginning of December. Do you have time at some point in this period? Thanks for the tips about getting it flat! My Problem is that I never used a CNC mill or any comparable tools so I would need someone to mill with me in order to not die. I would be eternally grateful if that was possible with you. Best, Lukas ________________________________ Von: CNC im Auftrag von Sebastian Bachmann Gesendet: Monday, October 28, 2019 2:38:11 PM An: Neigungsgruppe Zerspanungstechnik/CNC und Werkstatt allgemein Betreff: Re: [CNC] [Request] Designing/cutting Lab equipment for the real time visualization of DNA strand exchange On Mon, Oct 28, 2019 at 02:35:36PM +0100, Sebastian Bachmann wrote: > Hi! > > On Mon, Oct 28, 2019 at 01:26:03PM +0000, Lukas Lindorfer wrote: > > Hi Sebastian, > > > > thank you for your interest! The difference between the flow cells commonly used and the ones you are probably seeing Right now is that we do not work with whole cells but only with DNA strands and proteins in solution, so a little bit of shearing would not be too terrible. The holes are 4mm wide since they are optimized for those FPLC screws to make a wider range of applications possible: https://www.fishersci.co.uk/shop/products/peek-finger-tight-connector/11390052 > > Ohh okay, 4mm is not a problem and you can use plenty of tools to get a very > smooth finish even! > > > 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. > > > > What is critical is to have a really flat ground surface since TIRF microscopy is only practical in a range of about 200nm. I assume (but actually do not know) that the block to cut the cell out of would be rather flat to begin with. Is this the case? > > No, you can not assume that the blocks are flat. I would assume that you use > aluminium? In that case, the flatness is usually in the range of hundrets of ?m. Or less, obviously! There might be suppliers which sell precision stuff - but usually that is steel, which we can not handle in the CNC. > Milling the block flat, should reduce it - ideally you would surface grind it, > or lapp the block using a master plate (which we dont have). > What is the approximate surface flatness you require? > (see also here http://www.engineeringessentials.com/gdt/flatness/flatness.htm > for a definition of this measure) > > You can produce very flat surfaces by lapping three plates against each other. > Oxtools has some videos on that: https://www.youtube.com/watch?v=rHmsQEAx16o > > > > > > The model I sent still has some (a lot of) room for improvement ? Would you be interested in meeting at some point in the next weeks to talk about the cell? That way, I could give you some insights on what we know and need to know. > > Next week is very bad for me... I have time this week, and also wanted to visit > metalab and the CNC and see whats wrong with the VFD. > > Sebastian > > > > > Best, > > Lukas > > > > > > > > > > ________________________________ > > Von: CNC im Auftrag von Sebastian Bachmann > > Gesendet: Monday, October 28, 2019 2:12:20 PM > > An: Neigungsgruppe Zerspanungstechnik/CNC und Werkstatt allgemein > > Betreff: Re: [CNC] [Request] Designing/cutting Lab equipment for the real time visualization of DNA strand exchange > > > > 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 400?m. > > When looking at the screenshots, I suspect that the dimensions are in mm, hence > > the hole should be 500?m 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. > > > > > > > > > > > > > > > _______________________________________________ > > > CNC mailing list > > > CNC at lists.metalab.at > > > https://lists.metalab.at/mailman/listinfo/cnc > > > > > > _______________________________________________ > > CNC mailing list > > CNC at lists.metalab.at > > https://lists.metalab.at/mailman/listinfo/cnc > > > _______________________________________________ > > CNC mailing list > > CNC at lists.metalab.at > > https://lists.metalab.at/mailman/listinfo/cnc > > > _______________________________________________ > CNC mailing list > CNC at lists.metalab.at > https://lists.metalab.at/mailman/listinfo/cnc _______________________________________________ CNC mailing list CNC at lists.metalab.at https://lists.metalab.at/mailman/listinfo/cnc -------------- next part -------------- An HTML attachment was scrubbed... URL: From lukas.lindorfer at hotmail.com Tue Oct 29 09:27:01 2019 From: lukas.lindorfer at hotmail.com (Lukas Lindorfer) Date: Tue, 29 Oct 2019 08:27:01 +0000 Subject: [CNC] [Request] Designing/cutting Lab equipment for the real time visualization of DNA strand exchange In-Reply-To: References: <20191028131219.fsg4unegdxav4qgk@helios>, Message-ID: 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 if you have further questions. Best, Lukas ________________________________ Von: CNC im Auftrag von Armin Faltl Gesendet: Monday, October 28, 2019 2:27:12 PM An: 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 400?m. > When looking at the screenshots, I suspect that the dimensions are in mm, hence > the hole should be 500?m 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. >> >> > > >> _______________________________________________ >> CNC mailing list >> CNC at lists.metalab.at >> https://lists.metalab.at/mailman/listinfo/cnc > > _______________________________________________ > CNC mailing list > CNC at lists.metalab.at > https://lists.metalab.at/mailman/listinfo/cnc _______________________________________________ CNC mailing list CNC at lists.metalab.at https://lists.metalab.at/mailman/listinfo/cnc -------------- next part -------------- An HTML attachment was scrubbed... URL: From metalab at lightbug.at Wed Oct 30 22:17:36 2019 From: metalab at lightbug.at (ripper) Date: Wed, 30 Oct 2019 22:17:36 +0100 Subject: [CNC] Ausgeborgtes Werkzeug Message-ID: Hey! Um es kurz zu machen, wenn ihr Werkzeug aus dem Metalab ausgeborgt habt: Bringt das zur?ck, oida! Wir vermissen seit einiger Zeit die Flex und die Stichs?ge aus der HM. Mir ist egal wer das wof?r ausgeborgt hat, das Zeug soll einfach wieder im Lab auftauchen und nicht wie der Akkuschrauber letztes Jahr noch Monate weg sein... Danke und LG Michael -- ________________________________________ https://metalab.at/wiki/Benutzer:ripper -------------- next part -------------- A non-text attachment was scrubbed... Name: signature.asc Type: application/pgp-signature Size: 819 bytes Desc: OpenPGP digital signature URL: