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Cleaning sample lines after a clog

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JamesW

Contributor

Posts: 37

Joined: Tue Nov 21, 2017 6:59 am

Post Mon May 13, 2019 5:27 am

Cleaning sample lines after a clog

Dear All,

I wanted to ask everyone about how much they are able to clean and reuse a sample line (or sample capillary) after a clog. For example, if after a clog that completely prevented sample uptake and then flushing with a syringe, the line runs at 30ul/min but at increased pressure (say 13 up from 9) which does not drop after running cycles of water and wash solution, would you immediately change the line to a new one? Also do you have a set strategy to clean and reuse the partially clogged line?

Currently we flush several times with a syringe filled with water and try to run cycles of wash and water. It works most of the time but its pretty rare that we get a full month out of a line.

Any advice much appreciated.

Best,
James
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eganio

Contributor

Posts: 21

Joined: Wed Nov 07, 2018 8:00 pm

Post Mon May 13, 2019 2:36 pm

Re: Cleaning sample lines after a clog

Hi James,

One "quick fix" that usually works for us is to snip the ends of the capillary line (assuming you have enough length), since clogs generally tend to form at the junctures. If that doesn't help, it's probably worth making a new line if you got a clog in your old one, since the material cost of making a new one is easily dwarfed by the cost of risking losing portions of your precious samples because of clogging.

-Ed
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mleipold

Guru

Posts: 5796

Joined: Fri Nov 01, 2013 5:30 pm

Location: Stanford HIMC, CA, USA

Post Mon May 13, 2019 11:06 pm

Re: Cleaning sample lines after a clog

Hi James,

We mainly do try flushing the nebulizer capillaries (grounding nut to back of nebulizer), and most of the time, manually flushing the lines with a syringe does a decent job of clearing out clogs (along with running Wash Solution or 3% nitric, of course).

That said, if we're having trouble with a capillary, we just replace it. We buy 100ft spools of the tubing from Cole-Parmer rather than just rely on what we get from Fluidigm, so it's not too bad regarding cost or time. Since the screw fittings and such are reusable many many times over, you're only throwing out a few cents of tubing. In fact, we usually have 8-12 of the capillary kits assembled at any time: we usually wait for a few of them to get clogged, then do a bunch at once.


Mike
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JamesW

Contributor

Posts: 37

Joined: Tue Nov 21, 2017 6:59 am

Post Tue May 14, 2019 6:20 am

Re: Cleaning sample lines after a clog

Dear Ed and Mike,

Thanks for the information. For the tube cutting do you use a specific pipe cutting tool? We have a PTFE pipe cutter from Dolomite but it recommends using each cutting hole only once which would prove expensive over time.

Mike, I recently obtained some tubing* you recommended on another thread. However, I seem to be having trouble fitting it into the capillary kit. I can dissasemble and reassemble an old sample capillary kit using the tubing from fluidigm but if I try to use the Cole-parmer tubing it seems too narrow and just slides in and out of the assembled ferrule and clear sleeve without resistance. I'm not quite sure what the trouble is so any advice would be very helpful.

Best,
James

*Cole-Parmer
Ultramicrobore PTFE Tubing, 0.004"ID 0.016"OD, 100 ft/pack
Item# EW-06417-72
http://www.coleparmer.com/Product/Ultra ... m=06417-72
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mleipold

Guru

Posts: 5796

Joined: Fri Nov 01, 2013 5:30 pm

Location: Stanford HIMC, CA, USA

Post Tue May 14, 2019 2:52 pm

Re: Cleaning sample lines after a clog

Hi James,

I just checked our current spool: try the Cole-Parmer Item# EW-06417-74 (74, not 72). I think technically this means you'll have a 150um i.d. rather than 100um i.d.; the only real consequence is that your PSI pressure will be ~3-5psi rather than 7-10psi.

In short: we had the same issue you describe. After a few years of ordering the -72, we and several other labs noticed that the -72 was slipping out of the blue sleeve. We're pretty sure that this is a Cole-Parmer thing: while the specs on the -72 tubing didn't change, we had a good spool followed by a couple bad spools. We're pretty sure it was *not* a Fluidigm blue sleeve issue, as the -72 tubing was slipping out of blue sleeves and yellow ferrules we had used successfully before (and recutting blue sleeves and replacing yellow ferrules also didn't solve the problem). Also, using the same white plastic screw fittings didn't solve the problem.

A colleague tried ordering the -74 instead, and it's been working fine for us ever since.


Mike
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JamesW

Contributor

Posts: 37

Joined: Tue Nov 21, 2017 6:59 am

Post Wed May 15, 2019 4:01 am

Re: Cleaning sample lines after a clog

Hi Mike,

Thanks for that. Currently I am using the preassembled samples line kits (Product # 105922) with a clear sleeve but I will get a couple of blue sleeves when i get the new tubing in case it makes a difference. Does the wider bore of the tubing effect event rate or frequency of clogging?

Also, sorry one other quick question, how do you cut the tubes? Do you need a specific tool or can a sharp pair of scissors do the job without pinching the ends?

Best,
James
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mleipold

Guru

Posts: 5796

Joined: Fri Nov 01, 2013 5:30 pm

Location: Stanford HIMC, CA, USA

Post Wed May 15, 2019 6:12 am

Re: Cleaning sample lines after a clog

Hi James,

The slightly wider internal diameter doesn't seem to make much of a difference in clogging: we still have clogging in the PSI sip, at the grounding nut, and in the capillary.....we don't see a shift of clogging to the nebulizer, if that's what you're worried about.

We use razor blades: you need a really super sharp edge. Push the length of capillary through the blue sleeve until it peeks out the end just a bit, then slice through both the blue sleeve and the white capillary (take off a millimeter or so). Make the cut as orthogonal/flat as possible. The blue sleeve prevents the razor from crushing the capillary shut.


Mike
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JamesW

Contributor

Posts: 37

Joined: Tue Nov 21, 2017 6:59 am

Post Thu May 16, 2019 12:05 am

Re: Cleaning sample lines after a clog

Hi Mike,

Many thanks for the advice, I will give it a try.

Best,
James
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twightman

Participant

Posts: 18

Joined: Thu Oct 23, 2014 5:49 pm

Post Fri Sep 10, 2021 5:21 pm

Re: Cleaning sample lines after a clog

mleipold wrote:Hi James,

We mainly do try flushing the nebulizer capillaries (grounding nut to back of nebulizer), and most of the time, manually flushing the lines with a syringe does a decent job of clearing out clogs (along with running Wash Solution or 3% nitric, of course).

That said, if we're having trouble with a capillary, we just replace it. We buy 100ft spools of the tubing from Cole-Parmer rather than just rely on what we get from Fluidigm, so it's not too bad regarding cost or time. Since the screw fittings and such are reusable many many times over, you're only throwing out a few cents of tubing. In fact, we usually have 8-12 of the capillary kits assembled at any time: we usually wait for a few of them to get clogged, then do a bunch at once.


Mike



Hi Mike,
We are looking into possibly making our own capillaries and would like to know how you reuse the yellow ferrule and the metal piece that goes over that? For us as soon at that is crimped it makes both of these parts unusable. Thank you in advance for any insight you have.
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mleipold

Guru

Posts: 5796

Joined: Fri Nov 01, 2013 5:30 pm

Location: Stanford HIMC, CA, USA

Post Fri Sep 10, 2021 5:43 pm

Re: Cleaning sample lines after a clog

Hi Terry,

You can use your hands to pull the blue sheath tubing out of the yellow ferrule+silver ring. Then, take a pair of needle-nose pliers in one hand to grip the silver ring, and the fingers of your other hand to grip the yellow ferrule and pull apart. Don't use pliers on the yellow ferrule, as that will warp the plastic and make it useless....your fingers are just the right "grip" to pull without warping.

Then, take the pointy tip of a P200 pipette tip and push and twist lightly against the cap (pointy, non-flat) end of the yellow ferrule to open it back up a bit. This will allow you to get the blue sheath+white capillary back into the pointy end of the yellow ferrule and push until flush with the flat end of the ferrule.

Order of operation:
1. Use your hands to disassemble the blocked capillary setup into 2 white screw fittings (what goes into grounding nut and what goes into the back end of the square connector nut), 2 silver ring+yellow ferrules (still stuck together), 1 square connector nut with the nebulizer port screw fitting still attached, 2 blue sheaths, and the blocked capillary (which you throw away).

2. Pull apart the silver ring and yellow ferrule with your fingers and needle-nose pliers as described above.

3. Open up the pointy end of the yellow ferrule as described above.

4. Cut appropriate length of new capillary tubing (you can use the blocked piece as a measurement).

5. Push one end of new capillary tubing into a blue sheath until just peeking out. Slice a flush end of the capillary tubing+blue sheath with razor blade (needs to be sharp so you don't crush/crimp it).

6. Slide white screw fitting onto the end with the blue sheath until the sheath+capillary end is poking through. Slide on a silver ring (slightly tapered end pointing outward, away from the white screw fitting), then push on one of the widened yellow ferrules.

7. Make the end of the capillary+sheath flush with the flat end of the yellow ferrule, then take the square fitting and screw it on tightly. This treats it as an anvil to force the silver ring back onto the taper of the yellow ferrule to hold everything in place. Once you have forced it on there as tightly as you can, unscrew from the square fitting. If you push lightly on the blue sheath to move the silver ring+yellow ferrule slightly out of the white screw fitting, you should see that the capillary+sheath+ferrule+ring should move as one unit.

8. Do the same operation for the other end of the capillary. However, when screwing things into the square nut, do it gently until you feel it hit the end of the opening; do not tighten just yet. That should line up the capillary with the hole in the share nut; you should be able to push the end of the capillary through the hole and out the far end (nebulizer port). Once the appropriate length extends out, *then* tighten the back white screw firmly to clamp everything in place.


Please note: in our experience, the yellow ferrules can be reused multiple times. However, at some point, you will fatigue (irreversibly compress) the conical part of the yellow ferrule to where it will no longer clamp down on the sheath+capillary. At that point, you need to replace the yellow ferrule.


Hope this is clear,

Mike

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