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BSK-H Culture, Darkfield Microscopy, MS Patient

Posted: Sat 23 Apr 2016 1:22
by beneaththeskin
Hey all!

Just thought I would share some personal experimentation with y'all.

Here is a short video from one of my BSK cultures.
Incubated at 94.0F for 1 month, whole blood into BSK culture medium within a Vacutainer.

https://youtu.be/dH3fGVndMYo
Watch in HD 1080p!
Just like to share!

Very respectfully,
Forum member beneaththeskin

edit: Added that patient currently diagnosed with MS.

Re: BSK-H Culture, Darkfield Microscopy

Posted: Mon 25 Apr 2016 1:44
by lou
Gives me the creeps to watch these things swim around.

If a layperson can do this, it can be done. So, why isn't it being done?

Re: BSK-H Culture, Darkfield Microscopy

Posted: Mon 25 Apr 2016 10:24
by beneaththeskin
It can be done by a layperson, yes, however, I am not a layperson.

And it is extremely time/resource consuming. I think that is probably the main reason. It would otherwise be another tool useful to both clinicians and their patients.

Re: BSK-H Culture, Darkfield Microscopy

Posted: Mon 25 Apr 2016 13:15
by duncan
I don't think that the stated reason it is not embraced as a diagnostic tool is because it is time-intensive or expensive, but I could be mistaken.

My understanding is that it is not reliable enough. That there are claims that laymen or even clinicians/researchers using microscopy cannot replicate their findings in blind studies.

I hope I am wrong about this, because having an easy and reliable method for obtaining direct proof of Bb or other TBD's would be wonderful.

Are there published studies showing a high degree of reliability?

Re: BSK-H Culture, Darkfield Microscopy

Posted: Tue 26 Apr 2016 3:18
by beneaththeskin
Yea, not sure. I haven't looked into why it is not.

Also, you would still have to determine the species of whatever bacteria is in the culture, anyways.

Edit: For example, I am not willing to label the bacteria found with a name. I might state what I THINK it could be, based on other factors, but not specifically that this IS BB.

Re: BSK-H Culture, Darkfield Microscopy

Posted: Tue 26 Apr 2016 3:40
by ChronicLyme19
Wow, that's cool and horrifying at the same time. Send your culture to Dr. MacDonald to stain?

I've got some decent scopes in my lab and I've contemplated trying to do something like this to look for persisters. So you basically collect blood in EDTA vacutainer, then place in BSK culture medium and incubate at 94F? Then just take out the culture and place under the scope in darkfield? What was the original mag it was taken at?

Re: BSK-H Culture, Darkfield Microscopy

Posted: Wed 27 Apr 2016 2:09
by beneaththeskin
Yes, blood collected into EDTA vaucatiner.

Then, I inoculate 6.0cc BSK-H with 1.0cc whole blood from the collected samples.

I have two incubators running currently. One at 94.0F and one at 100.0F.

Some cultures are of whole blood. Some are of plasma layer after centrifugation, and some are of erythrocyte layer after centrifugation.

Just experimenting with what works for my culture methods.