Bioprinting is printing with biological materials. Think of it as 3D printing, but with squishier ingredients! There's a lot of work being done at research labs and big companies like Organovo on print human tissues and human organs, with an eye towards drug testing, and transplantation into humans. Check out these amazing TED talks by Anthony Atala, for example:
Anthony Atala: Growing new organs
Anthony Atala: Printing a human kidney
All this sounds incredible complex, but the fact is that the basic technologies are very accessible - it's all based on inkjet and/or 3D printing! So a bunch of us at BioCurious decided we wanted to play around with this technology ourselves - and the BioPrinter Community Project was born! (Come join us, every Thursday evening at BioCurious!)
We wrote this instructable in part to document our project for our fellow Citizen Scientists in the DIYbio community, so it's getting a little long. For those who want a quick 1-minute intro, you may want to check this little video:
Anthony Atala: Growing new organs
Anthony Atala: Printing a human kidney
All this sounds incredible complex, but the fact is that the basic technologies are very accessible - it's all based on inkjet and/or 3D printing! So a bunch of us at BioCurious decided we wanted to play around with this technology ourselves - and the BioPrinter Community Project was born! (Come join us, every Thursday evening at BioCurious!)
We wrote this instructable in part to document our project for our fellow Citizen Scientists in the DIYbio community, so it's getting a little long. For those who want a quick 1-minute intro, you may want to check this little video:
Remove these ads by
Signing UpStep 1: Hacking an old inkjet printer to print biomaterials
We started out by messing around with an old inkjet printer that we literally saved from a sidewalk somewhere. There's already plenty of interesting things you can do with an low-end off-the-shelf inkjet printer, but they do have some limitations, which we'll get into in the next Step (or skip straight to Step 3 for how we built our own bioprinter from scratch, that you can see in the first picture above).
Undressing the Printer
We disassembled an abandoned HP 5150 inkjet printer for use as a bioprinter. Just rip off all the plastic covers you can find, but make sure you can still operate the reset buttons etc. on the front panel.
There's a little momentary switch that senses whether the cover is open. After you've ripped off the cover, you will need to close that switch to be able to print anything. You can just press it down by hand or with a toothpick, glue a little piece of plastic or a screw onto it to make it easier to depress, solder on an on/off switch, or even just bridge it with a piece of wire (so it thinks the cover is always closed).
There's also a momentary switch inside the paper handling mechanism that senses whether paper has been loaded. You may need to rewire that one as well is you plan to mess with the paper handling.
Once you've got your printer all undressed, and figured out how to activate the cover-closed switch - print something! It'll be a good test to see if you've knocked something loose and whether you can still operate all the buttons. Plus, it's just darn cool to see an inkjet printer do its job up-close. Keep your fingers out of the way though - those cartridges move fast!
Cartridges Prep
Next task is prying open an ink cartridges, so you can fill them with something more interesting. We will be using brand new cartridges for the printer, because old cartridges tend to have residual ink clogging their nozzles. However, you might want to practice opening expired cartridges first. Wouldn't hurt to do a test print, and flush the print head, if your printer software provides that option.
After taking off any labels, you'll see that most cartridges have a lid that's glued into place, usually with some small vent holes to let air in as ink flows out. If you run an Exacto knife or box cutter along the seam long enough, you should be able to cut through some of the glue, and pop the lid off fairly quickly. WARNING – boxcutters are SHARP! If you're a kid, get some responsible adult to help you with this! If you're an adult - consider protecting your hand with a knife-proof glove, and/or clamping down the cartridge in a vise while you're doing this.
Alternatively, you can cut off the lid using your favorite power tool. *Much* easier, but not quite as clean. Keep in mind that you may need to be able to put the cartridge back together again, to get it to fit into the cartridge holder. If anyone knows of an easier way to open inkjet cartridges, post something in the comments! Maybe there's some solvent that will loosen up the glue without dissolving the cartridge itself?
Once the cartridge is open, you'll see that the entire ink reservoir is taken up by a little sponge that holds the ink in place. Color cartridges have separate ink compartments with their own sponges (typically Cyan-Magenta-Yellow, not Red-Green-Blue, since printing colors on white paper is a subtractive color process). You can squeeze out any remaining ink and keep it for later experiments (e.g. for some paper chromatography). Next, rinse, rinse, and rinse some more with distilled or deionized water-we don’t want to clog the print head with mineral residue. We want to get all that ink out of there, so it doesn't interfere with our experiments.
Once the ink reservoir is completely clean, fill it partway with distilled or deionized water, put the lid back on, pop it back into the cartridge holder, and print something. Chances are, no matter how well you rinsed, you'll still see some residual ink coming out. Just keep printing until it's printing clear, and then print some more.
We discovered that our cartridge actually slowly leaks through the print head when you fill it completely with water, but it does not leak when you only put in a small amount. That actually provides a very convenient way to flush out the print head, without having to print.
Filling the Cartridges
At this point, what you want to put in the ink cartridge is entirely up to your imagination! We started out by putting some green food coloring in the formerly black ink cartridge, and printed out a test page to show that it worked. You could also try filling up the color cartridge with fluorescent dyes of your choice, and print out you own blacklight posters. Or, put some invisible ink in the black cartridge, and print invisible messages to your friends.
As our first real "bioprinting" experiment, we wanted to start with something simple, instead of jumping straight into printing with live cells. We decided to print with a solution of arabinose onto filter paper. Then we cut out the filter paper, and put it onto an agarose plate on which we had grown a lawn of E. coli that we had engineered to carry the pGLO plasmid. This plasmid carries the Green Fluorescent Protein (GFP), under control of an arabinose-sensitive promoter. (Stay tuned for an instructable on how to make your own GFP-expressing E. coli).
As a result, wherever we had printed arabinose on the filter paper, we now saw the E. coli light up green under UV light! Note that the beauty of this experiment lies in its simplicity: we only had to print with a simple sugar solution, rather than with bulky live cells; and we were printing on paper, so we didn't even have to change the paper handling machinery. You could also try printing with antibiotics, or even proteins, such as enzymes or growth factors.
The second-to-last image above shows our first test print, where we has printed arabinose over half of the filter paper - and half of the plate lights up under UV light. In the second image, we had printed the BioCurious "eyeball" logo. Success! Unfortunately the sharpness of the image definitely leaves much to be desired. Presumably, the arabinose tends to diffuse through the filter paper, which smears out the printed pattern. We should be able to do much better by printing directly on the agarose.
Undressing the Printer
We disassembled an abandoned HP 5150 inkjet printer for use as a bioprinter. Just rip off all the plastic covers you can find, but make sure you can still operate the reset buttons etc. on the front panel.
There's a little momentary switch that senses whether the cover is open. After you've ripped off the cover, you will need to close that switch to be able to print anything. You can just press it down by hand or with a toothpick, glue a little piece of plastic or a screw onto it to make it easier to depress, solder on an on/off switch, or even just bridge it with a piece of wire (so it thinks the cover is always closed).
There's also a momentary switch inside the paper handling mechanism that senses whether paper has been loaded. You may need to rewire that one as well is you plan to mess with the paper handling.
Once you've got your printer all undressed, and figured out how to activate the cover-closed switch - print something! It'll be a good test to see if you've knocked something loose and whether you can still operate all the buttons. Plus, it's just darn cool to see an inkjet printer do its job up-close. Keep your fingers out of the way though - those cartridges move fast!
Cartridges Prep
Next task is prying open an ink cartridges, so you can fill them with something more interesting. We will be using brand new cartridges for the printer, because old cartridges tend to have residual ink clogging their nozzles. However, you might want to practice opening expired cartridges first. Wouldn't hurt to do a test print, and flush the print head, if your printer software provides that option.
After taking off any labels, you'll see that most cartridges have a lid that's glued into place, usually with some small vent holes to let air in as ink flows out. If you run an Exacto knife or box cutter along the seam long enough, you should be able to cut through some of the glue, and pop the lid off fairly quickly. WARNING – boxcutters are SHARP! If you're a kid, get some responsible adult to help you with this! If you're an adult - consider protecting your hand with a knife-proof glove, and/or clamping down the cartridge in a vise while you're doing this.
Alternatively, you can cut off the lid using your favorite power tool. *Much* easier, but not quite as clean. Keep in mind that you may need to be able to put the cartridge back together again, to get it to fit into the cartridge holder. If anyone knows of an easier way to open inkjet cartridges, post something in the comments! Maybe there's some solvent that will loosen up the glue without dissolving the cartridge itself?
Once the cartridge is open, you'll see that the entire ink reservoir is taken up by a little sponge that holds the ink in place. Color cartridges have separate ink compartments with their own sponges (typically Cyan-Magenta-Yellow, not Red-Green-Blue, since printing colors on white paper is a subtractive color process). You can squeeze out any remaining ink and keep it for later experiments (e.g. for some paper chromatography). Next, rinse, rinse, and rinse some more with distilled or deionized water-we don’t want to clog the print head with mineral residue. We want to get all that ink out of there, so it doesn't interfere with our experiments.
Once the ink reservoir is completely clean, fill it partway with distilled or deionized water, put the lid back on, pop it back into the cartridge holder, and print something. Chances are, no matter how well you rinsed, you'll still see some residual ink coming out. Just keep printing until it's printing clear, and then print some more.
We discovered that our cartridge actually slowly leaks through the print head when you fill it completely with water, but it does not leak when you only put in a small amount. That actually provides a very convenient way to flush out the print head, without having to print.
Filling the Cartridges
At this point, what you want to put in the ink cartridge is entirely up to your imagination! We started out by putting some green food coloring in the formerly black ink cartridge, and printed out a test page to show that it worked. You could also try filling up the color cartridge with fluorescent dyes of your choice, and print out you own blacklight posters. Or, put some invisible ink in the black cartridge, and print invisible messages to your friends.
As our first real "bioprinting" experiment, we wanted to start with something simple, instead of jumping straight into printing with live cells. We decided to print with a solution of arabinose onto filter paper. Then we cut out the filter paper, and put it onto an agarose plate on which we had grown a lawn of E. coli that we had engineered to carry the pGLO plasmid. This plasmid carries the Green Fluorescent Protein (GFP), under control of an arabinose-sensitive promoter. (Stay tuned for an instructable on how to make your own GFP-expressing E. coli).
As a result, wherever we had printed arabinose on the filter paper, we now saw the E. coli light up green under UV light! Note that the beauty of this experiment lies in its simplicity: we only had to print with a simple sugar solution, rather than with bulky live cells; and we were printing on paper, so we didn't even have to change the paper handling machinery. You could also try printing with antibiotics, or even proteins, such as enzymes or growth factors.
The second-to-last image above shows our first test print, where we has printed arabinose over half of the filter paper - and half of the plate lights up under UV light. In the second image, we had printed the BioCurious "eyeball" logo. Success! Unfortunately the sharpness of the image definitely leaves much to be desired. Presumably, the arabinose tends to diffuse through the filter paper, which smears out the printed pattern. We should be able to do much better by printing directly on the agarose.
Visit Our Store »
Go Pro Today »
Also would it be possible to convert this in to a 3d printer? Just wondering.
Thank you for your time.
thanks for sharing your work!
Do you know about something like this with Epson printers?
I think they are based on Piezo not thermal technology, could they be used for a wider range of materials?
"In our study, we comprehensively studied changes in heat shock protein expression and cell membrane morphogenesis in printed mammalian cells with thermal inkjet printers. The heat shock protein expression of the printed cells has minor difference between the untreated cells and lower than manually heated cells."
http://etd.lib.clemson.edu/documents/1239894674/Cui_clemson_0050D_10011.pdf
http://onlinelibrary.wiley.com/doi/10.1002/bit.22762/abstract
As for printing patterns with yeast cells, where do I start looking for information on the size of something like brewers yeast cells?
And, what size or type of filter will contain yeast cells while allowing a nutrient fluid to pass back and forth?
Thanks for any help you can give this newbie.
DIY-Guy
Human cells can vary a lot in size, depending on the type of cell. Human egg cells are >100 µm, while sperm has a diameter of only around 5 µm. Typical animal cells are quite a bit bigger than yeast cells though - let's say 20-50 µm.
Here's the corresponding table for "a bacterium". Consensus: ~1 µm, with a range of 0.2-5 µm.
Lastly, here's a really good animation of relevant sizes in biology.
Filter sterilization is typically done with a 0.2 - 0.45 µm filter, but keep in mind that viruses and phages will pass through such a filter. If you want to separate yeast cells from the supernatant, you should be able to get away with a 1 µm or even coarser filter, but 0.2 or 0.45 µm may be easier to find.
For everyone who is interested in buying an InkShield kit, I plan to have them available in my store again mid March.
Or bubblegum?
However, I doubt we'd be able to find a CIS kit for this particular special-purpose HP cartridge that we can drive with the InkShield. We could easily adapt one to our purposes, but that would involve opening the cartridge anyway.
Typically, we're only be printing very small volumes that need to be prepared fresh every time. So until we come up with some applications where we're printing relatively large volume (i.e. >5 mL or so) 3D structures, a continuous ink kit would be overkill.
Most ink refill kits just inject ink through one of the vent holes in the cartridge with a big blunt-needled syringe, or even include a tiny hand drill to make your own hole in the lid. You may be able to flush enough water in and out through that hole with a syringe, to then put different colors in the color cartridge. Get some cyan, magenta and yellow highlighters, squeeze the ink into the inkjet cartridge, and print your own black light posters!
I see with your 3rd pic that you've (*joking*) managed to print a human heart!
Makes me wonder if you are working on you're own replacement parts, or someone to help around the house.
Let me know when he/she is finished! Do you plan to licence him/her under the GNU GPL or BDS licences? Or is this to be a closed-source person? *grin*
Seriously, I saw a project like this on PBS several years ago, talking about printing scaffolding upon which human cells could grow, and lamented that they had copyrighted the modifications they'd made to the print-cartridge. Though I figured that this sort of stuff was probably out of the realm of possibilities for the hobbyist.
Glad to see you've proven me wrong!
Speaking of human hearts - as I mentioned we don't have any plans to use human or animal cells in the foreseeable future, but we did have have serious plans to make a decellularized "ghost heart" for Halloween last year! We actually did some test runs trying out different protocols to decellularize some chicken hearts using various methods. Haven't gotten around to scaling it up yet, but one of our members does now have two pig hearts in his freezer :-D
Maybe we'll get around to that project again for next Halloween - we'll post an instructable if we do!
remember the OLDER the inkjet the better. new and or higher end inkjets have 2 and even 1 picoliter nozzles meaning whatever bio matter you put in their will probably just clog the nozzles up. so think OLDER.
Here is an ebay example. 110773910917 those come empty and are designed to be refilled so very friendly to this kind of project.
also CIS or Continuous Inkflow Systems. although they usually come "full"
Of course, sufficiently old inkjets are getting harder and harder to get, and the cartridges will probably go our of production as well. The other issue is finding drivers for them, and especially trying to modify drivers (or even write your own!) to get more control over what the printer does.
Using an older 300dpi inkjet is definitely a viable option for bioprinting, especially if you manage to get your hands on an a printer like the HP 500, where people have already figure out a lot of the details.
We just felt it was easier, and had more growth potential, to start with a clean slate...
Jose
Little Devices @ MIT
I don't seem to be able to log in to TechHive, so I'll just post my response here:
Obviously, we are *not* using any kind of harmful bacteria. And yes, DO try this at home! Whip up a yeast culture from a packet of dry baker's yeast, and see if you can print with that. Or get one of the genetic engineering kits from Carolina, make your own mutant glowing bacteria, and print with those.
Have fun! Build something! Learn science by doing!
GM