Hello,
I need to make organotypic slices using a Leice VT1200S vibratome. I cannot add antibiotics to the culture medium, so the slicing procedure needs to be done under sterile conditions. The vibratome is too big to fit inside a tissue culture hood.
All ideas welcome!
When we got our vibratome I also was looking for a protective hood (https://www.researchgate.net/post/Protective-hood-for-vibratome-work-with-patient-material). Since we could use antibiotics, we decided not to use one in the end. The problem was the dimension of the device. But, as far as I remember, these were the discussed options:
- Customized PCR hood from Mystaire (the quote was 5000$, so quite pricey)
- One of these cabinets had the right inner dimensions (barely): https://www.nuaire.com/biosafety-cabinet-nu-545 (but you need a lot of space)
- Self-made with HEPA air filter. Drawback: no real air barrier, so it would not be 100% sterile. And you need a skilled person for constructing this.
None of those ideas were ideal, which is why we did not pursue this further... But perhaps there are some new biosafety cabinets out now, which have the right dimensions.
Good luck!
PS: As a side not, since the organs for the slices come out of animals, they will not be sterile anyway
Hi Tamara
I used to make organotypic brain slices on a vibratome, also sometimes without antibiotics. I would use the same equipment and procedure as when using antibiotics, and surprisingly few cultures would get contaminated (I would say less than 10%).
Compared to how disruptive/expensive/impractical it may be to place the vibratome in a designated hood (As Suckert Theresa mentions above), I would suggest you try your normal approach first. Probably the use of a face mask during preparations is a good idea, in case you need to talk or cough.
Can I ask why you cannot use antibiotics; I know many people discourage it for various reasons, but in many cases you can readily get around possible confounders with easy control experiments? Also, while I did not do a systematic comparison, I never noticed obvious differences (in electrophysiological recordings) between cells from cultures with or without antibiotics (Pen/strep, which I would commonly use at 0.5x of recommended).
Best regards
Jan
Jan Tønnesen , thank you for your response. It is good to know that you did not get contamination often!
The reason I cannot use antibiotics is because they disrupt the spontaneous activity of developing cortical networks.
Best wishes,
Tamara
Hey Tamara,
I hope this answer isn't too late. Although we use antibiotics in our medium, but we also tried to reduce contamination by all different means. We always wear masks when dealing with the brains. We also dip the whole head before harvesting the brain in ethanol. We put the vibratome in a room where not so many people get in, and we clean everything we use of course with 70 then 100% ethanol.
Aya Alserw Thank you for your answer. I will try dipping the head in ethanol as well.