DL118 - Luminex 100 Flow Cytometry Analyser Teardown
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- Опубликовано: 7 апр 2017
- In this video i teardown a Luminex 100 Flow Cytometry Analyser. Used in bio-medical sciences.
en.wikipedia.org/wiki/Flow_cy...
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I'm an immunologist who's worked a lot with flow cytometers. Thanks for the great teardown! Just to say that the glass vial you broke is called a flow cell, and that needle going up into the base of it is where the actual sample is injected into a stream of sheath fluid flowing around it (so this particular machine actually has the sample flowing upwards through the flow cell). As the diameter of the channel decreases the fluid velocity increases according to Bernoulli's principle and the sample stream is stretched out into an extremely well defined shape, forcing the microbeads into single file with a consistent alignment with the optical components. This is called "hydrodynamic focusing".
Thanks for the explanation of the flow cell and needle arrangement.
DextersLab2013 No worries! It's always cool when you, mike, dave etc. tear down something that I've used as part of my work and i get to finally see inside it.
Interestingly, in most flow cytomteters they use a sheath fluid to flow around the injection needle, exploiting laminar flow to help keep the shape linear as it is stretched out, cutting turbulence and enhancing the single file stretching. Some use a needle within a needle setup, others just flow it carefully and uniformly through a tight space
From me? No sorry it was all sent into recycling after the teardown.
The acquisition board reminds me a bit of a digital sampling oscilloscope.
Those Coherent Compass lasers can sometimes take a couple of minutes to start - they have TECs on the diode, cavity and KTP to optimise the output.
The one I have has the most bizarre circuitry I've ever seen in the controller - you'd think they'd use an MCU, but it has a really wierd collection of PLDs and digital pots, and a strange relay in a very compact and repair-unfriendly collection of PCBs. Your one is probably rather more recent so YMMV
Internal construction of the laser is quite interesting - they are aligned with a jig, and heaters in the baseplate used to melt solder to lock everything into position.
i opened the lid on the controller and it's completely different to the designs on sams laser faq, very tempted to open the laser head though!
You should definitely take the laser module apart.
Looking forward to power on the lasers and the opening of the photo-multiplier module, without any glass break.
I did get into the PMT module in the end but it's entirely potted, there's not much to see which is why i didn't include it in the video.
500 shades of spheres wait I thought it was 50 shades of spheres
Great video. Awesome Hamamatsu stuff. Thank you for sharing.
Thanks Max!
I'm sure there's an interlock circuit on the laser controller. probably need to figure out how to bypass it for them to come on
yea it's all detailed on sam's laser faq: www.repairfaq.org/sam/laserscl.htm#sclc215
I so badly wanted to take the hex driver that was in the compartment and tell you to try and see if it fit those other screws you resorted to removing with a torx. ;p
US stuff tends to be imperial - you need a key set in 64ths some only do 32nds.
thanks, i'll have to source some
Hey a topic I know very very very well. I actually worked as a conjugation chemist designing the dye reagents for use in flow cytometers. This is a very budget model, if you want to see the higher end check out a Becton Dickinson (BD) LSR II or LSRFortessa. Those can cost north of $500k when new and well configured with software, filters, over a dozen PMTs, etc. The optics on these more advanced systems gets REALLY REALLY complex with you being able to detect over a dozen signals at one time. Each laser has its own bank of up to 8 different PMTs with associated filters. The signal enters each chamber and using a combination of low pass filtering mirrors and band pass filters, they are able to have only the filters that should be receiving a signal receive light. What is more interesting is each set of PMTs is exclusive to its own laser, meaning a dye which is excited by the blue laser will only have its signal on the blue laser bank of PMTs, even if there is a similar color of emission on the yellow laser channel. These are very complex and VERY powerful devices.
In your setup, the mirrors likely serve as low pass filters. Allowing the light through that is below the wavelength desires. Though I don't see it in this system, it is very common to inclue bandpass filters after the LP filters to ensure only the desired light enters the detectors.
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If there is anything you want me to clarify or any question you have about the dyes/conjugates which are used in these systems feel free to ask.
Thanks for the comments! Yes all the mirrors had different coatings on, not sure if i mentioned that in the video though! I wouldn't mind knowing the rough cost of the machine when new if you have some idea?
I haven't found a source for the price of the Luminex 100, but the 200 (a related system) runs around $60K base cost. This likely didn't include software ($12K) or various options which would raise the price more.
i am guessing the reagents are not cheap either?! Thanks for the info, it's always nice to know how much something cost.
The reagents aren't too bad on a per test scale but the costs can add up quick. For example, a basic reagent might run you as cheap as US$2/test and all of the reagents needed for the test might run around US$10-50/test. Where it gets a expensive is that a single experiment, including all of the requisite controls and calibrations could run between 20 and 100 tests, with large complex experiments requiring even more. This results in a typical price of US$200-US$5000 for a single experiment, not including operator time costs or instrument costs.
On the flip-side, the information you can get from a Flow Cytometer is remarkably powerful and really can't be achieved any other way. You can break down cellular populations by cell type, to see which cells express which proteins, how treating cells in certain ways can change the expression of said proteins, etc. The power comes from how you can "drill down" within populations into sub-populations and sub-sub-populations, etc. Much of the modern knowledge on immunology and in vivo cellular protein transcription has come via flow cytometry. This is useful in things like cancer tracking (monitoring cells for cancer specific protein synthesis), autoimmune disorders, evolutionary biology, genetics and more.
W00t Flow Cytometry!
I've always wondered how 'safe' it is to take one of these apart. In the biological sense, seeing as you have no idea what chemicals or bio waste is in there. A lot of the ones for sale will have a warning saying they could possibly have biological hazards in them, or other ambiguous language such as "it is the buyers responsibility to safely clean the unit". Interesting to see this one still had liquid in it.
I was thinking about it too and TBH i do wonder in a professional environment that the machine would probably be cleaned and flushed after each assay so it's ready for the next
Laser modules probably don"t work due to safety interlocks, especially for class 3B lasers. Work out how to defeat the interlocks and you most likely will get them to work.
looks like a budget unit costs around $50k but a 2 laser one is more like $100k but the info I saw said prices have come way down so that one was possibly more like $150k.
nice, thanks for doing some digging!
ooh that was a good one! :D
Thanks!
love you always tear down competitors products to products that my company makes. Wonder if you'll ever open one of ours. ^_^ But yeah, class I is just as the consumer is intended to use it.... we mostly use 3B, but have some 4 in some of our instruments. FDA regulations require us to interlock laser devices. I'm not sure about that manufacturer, but the one that we get our modules from, there is actually a enable waveform required in that is controlled by the processor, so that if the processor fails it disables, and if any interlocks are broken (except if in service over-ride) the processor will disable the waveform. I call it a wave form, but its basically like a pwm signal. But either way, the laser controller will have to have some sort of enable line in one way shape or another.
it's just random picks from the selection of equipment on ebay, there are a lot of manufacturers to choose from!!
indeed. Anyhow, if ever in Miami and want to see the assembly of blood and particle counters/sizers, let me know. Sadly no videos allowed.. you know how it goes.
Thanks for the offer, maybe one day!
I didn't realize we had any companies down here that made that kind of analytic equipment. Hmm.
don't kill the coherent you can find the module manual online
Open the laser.
Can you not get more data on the green laser and the driver module? I would guess being a class 3b there may have been safety interlocks or somesuch preventing the unit from firing until conditions were met etc. Just a wild guess....
There's full details over at sams laser faq so i followed those details, i'll double check my findings before i open it but it's not looking good. TBH i do have a working 20mw green laser anyway so i am not too fussed if it's toast
show off ;-) lol
Where did you got the unit for from?
Just bought off ebay.