centiSperm DIY Protocol: Sperm Desalination
Goals: To electroporate whole genomic DNA from a centipede into the center of human sperm.
Sperm Desalination Rinsing the sperm is meant to remove the salt from the sample. This is to reduce the electrocution risk to the sperm during electroporation. The electroporator gives high voltage millisecond pulses and if the sperm is in a salty seminal fluid medium, the electroporator will just zap the sperm to death. The process is much like being a human washing machine. The actual process is slow, boring and repetitive: spin, drain liquids, agitation rinse, spin, drain liquids, agitation rinse, spin, drain liquids, agitate into a sort of gel. Matteo was heard to say, “This is why I stopped being a scientist.”
A - Obtain Sample
B - Put sperm in tubes
I had very few tubes with me. I thought to use disposable pipettes but I got worried about sperm spray because the tubes would have no tops. I bought hypodermic needles at the Wallgreens to use as sterile tubes. I figured that the syringes could be spun and the heavier sperm would deposit the pellet onto the black rubber plunger and then I could push out the liquid (supernatant) and suck in new liquid to resuspend into (vibrating agitation rinse). I used electrical tape to secure the needle closed. I used the screw-on.twist-off luer lock for as the cap for the syringe tube. This made accidental sharps sperm injection much less likely! I also taped the plunger closed so it wouldn’t fly off the fan releasing sperm in a circular spew pattern. I left tape slack so I could still plunge without having to worry about coating the place with a thin film of half rinsed ejaculate.
C – Spin to make pellet (natant)
I was going to buy a hand-powered centrifuge but I thought better of it. I tried to make due with what I had brought and immediate materials. Although a DIY handheld grinder centrifuge was beta tested. The results were dangerous. David Bowen and I put the grinder in a vice and stepped out of the shop to plug it in. The attachment took air, collided with the vice and self destructed sending shrapnel in every direction. We were nearly maimed in the face by a projectile syringe.
The DIY box fan centrifuge was the answer. I drilled holes in the plastic fan blades and zip tied the syringes to the blades of the fan. It took a little time to cut them off between each cycle but we did see a thin white line of sperm solids around the plunger after each spin.
Spin for about 3 minutes
D - Throw away supernatant (liquid)
Plunge the seminal fluid supernatant into the bleach bath. A word on DIY Biosafety: When working with human fluids, it is good to have art as a shield from the health department. Art is not usually considered to be stringently regulated, as are federally funded laboratories and biobased economy manufacturing processing plants. Using somewhat dangerous materials in performance art or DIY labs is generally ignored. Not that the health department doesn't have jurisdiction. There are just some funny ideas about art. It is generally considered to be sort of like a party in the ambi-bathroom of a good afterhours club. Art is a sleazy urinal cake. So, the mythos of general consensus art freedom from oversight is sort of a mix between fourth amendment freedom of expression issues and that soldier of fortune motto ‘kill em all, let god sort em out’. This is the attitude towards the arts and artist’s in general society. This is to say that the impact of art is zero to nil in the public eye and the desperate narcissism of most artists doesn’t hinder this vision. Proclaiming yourself an artist is like wearing a classless green translucent plastic masochist visor on your head. Artists signify as frivolous, expendable and hence deregulated. It is sort of an unsaid authoritative death wish hex that translates to freedom from funding and freedom to self regulate. Nonetheless, I used a water purification pill good for 25 liters in a bin as a bleach bath for both used fluids and plastic disposable trash. No harm in sterilizing the shit out of stuff, neh?
Just suck distilled water into the syringe. Use the syringe plunger to fill the syringe with distilled water. Don't forget to leur lock the sealed needle back on the syringe so there is minimal leakage. F – Vortex Resuspend sperm in distilled water by vortexing A vortex is a great home toy to invest in. It is an industrial strength vibrating twiwrler. Its major use in a lab is for resuspending tacky solids stuck to the bottom of tubes after centrifugation. It can also be used a fun drink mixer. It does make that tell tale tornado in fluids pressed onto the nob of the vortex. The strength and RPM is well beyond the standard vibrator so there are other more potent uses for a home vortex related to: massage, deep tissues, and earthshaking stimulation possibilities. Well I left my home vortex at home. The DIY vortex for resuspension was simply to attach two syringe wrappers to a handheld sander and let the sander shake and vibrate the sperm solids back into solution with the saltless distilled water rinse. Sometimes I would press the hypodermics onto the sander surface to see the liquids really shaking and splashing and spinning and waving about. It worked! It was a little foamy, frothy for professional use, but the saucy was scenic. G - Spin to make pellet (natant Spin for about 5 minutes H - Throw away supernatant (liquid) Plunge the distilled water seminal rinse supernatant into the bleach bath I - Replace distilled water with ice cold distilled water J - Vortex Just another jot on foam and happenstance.
There was some leakage of sperm from inside the hypodermics to outside on the surface. Good thing this lab was mostly stag. Gloves were offered and the dangers explained. But, during vortexing, the shaking and vibrating of the lost droplets of sperm appeared as foam. This again confirms that sperm, left out and agitated by waves, does make a foam! This foam is thick, meringue-like, white, bubbly, a shave cream consistency, a lather of leaky fluffed goo spread. Its an epic yuck factor emollient. It’s also proof of concept in the foam as castrato sperm meets ocean vortex theory of Aphrodite’s origin. K - Spin to make pellet L - Throw away supernatant M - Replace ice cold distilled water with ice cold Glycerol I bought glycerine at an uptown health and beauty herb pharmacopeia called Pharmaca in PaloAlto. It was the cheapest item in the storehouse of flower dew remedies and herbal tinctures. Glycerol is 10% glycerine. That is 9 parts distilled water and one part glycerine. A solution of glycerol is often used to keep just to keep mixtures suspended in a smooth, moisturizing solution. It is also a sort of preservative. For vegetarians, there is plant-based glycerol. Glycerol is used in lots of stuff we use every day. You can find it in shampoo, water based sex lubricants and in this case, a suspension of sperm. If well screened, young sperm suspension with a little fragrance, perhaps lavender, might make a very good face and cuticle cream for the boutique beauty product market.
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