bone saw + establishing the bureau of microscopic investigation

Stuart informed me today that I should be keeping a lab book of my protocols, experiments and results, and so this research blog will serve as a type of Lab Book. In fact, this gives me ideas for a more creative presentation of my research report in the end: to organize and aestheticize it like a Lab Book.

For starters, I've created these lab log cards, based on traditional forensic/police cards. The invention of the Bureau of Microscopic Investigation is mine:

These cards will be completed each time I work in the lab and will be included in a final 'lab book' report. 

Photo by Moe Beitiks.

Yesterday I borrowed the electric bone saw from the morgue and used it to cut a huge cow bone in half, so that I could scrape out the bone marrow and some of the compact bone tissue. My tour of the morgue was interesting, including sightings of various dismembered human body parts on steel stretcher tables, some visible only as heaps under sheets and some open for plain viewing. I did see a human head, with sections removed, including the skin. Once I had my bone saw in hand, I was quickly off to the lab to do some living science.

Taking cells directly from a fresh, raw bone versus buying a cell line from a laboratory supply company is called primary sourcing. Primary sourcing is considerably cheaper (most cell lines sell for around $500) albeit fraught with problems. Contamination is one common problem. If a cell culture is contaminated with bacteria, fungus, etc it becomes unusable. I am happy to report, after having Stuart inspect my new bone cell cultures, that they are not contaminated. However, he says he doesn't really see many viable cells, either. That means all of the cells I did get from the marrow have either died or are still alive but not adhering to the flask, meaning they won't be able to grow and will die. I still have hope, though. He says it's hard to tell for at least a few days. I only gave mine 24 hours before I was inspecting the hell out of them. I saw many interesting things under the microscope, including a whole bunch of little tiny black dots, which seemed to have an aura. I was afraid that these were bacteria, but it turns out, Stuart tells me, they are red blood cells. Millions of red blood cells. He says they will eventually all die because they don't have much of a life cycle. So, in the next couple of days, I'll hope for some viable bone cells to adhere themselves to the flask, and then I can grow many from them.

Photo by Moe Beitiks.

I was very lucky to have no contamination, considering I was digging in a dirty old bone after sawing through it with an old morgue bone saw, and it was my first time really getting under the hood in the lab. Burning bone stinks of burning hair, by the way. Ionat was having a hard time with the smell. Stuart told me that I should always test the cell medium (DMEM) for contamination before putting a new culture in it. This means putting the plain medium in a dish by itself for a few days in the incubator to see if any bacteria grows in it. If it does, the medium is contaminated and needs to be filtered before being used. I'm going to do that today before I leave, though it looks like I don't have contamination anyway. However, sometimes contamination can take a few days to show up.

Photo by Moe Beitiks.

Another thing I should have done first, Stuart tells me, is to put the cells not directly into the bone soup that I mixed but instead just directly into plain DMEM. The reason for this is that the bone soup I made is specifically for osteoblast differentiation, meaning I am trying to force the bone cells to become one type of cell (an osteoblast) and if they aren't liking those conditions, the cells will just die. All cells like plain DMEM, so I could just put my primary source culture in that first and just wait and see what grows, then decide if I want to put anything in my precious bone soup.

The eucalyptus bark and the tree that I collected it from.

Last night I gathered eucalyptus bark from the riverside and boiled it in one of Janet's large cauldron-like pots to create my first natural dye bath which I hope to use as a tissue stain.

My eucalyptus dye pot. Kitchen lab!

I asked around today as well, about staining a living culture. Apparently it isn't routinely done - most cultures have to be fixed first with formaldehyde although it is possible with some types of fluorescent stain. I'm specifically interested in using the stains for medicinal purposes, such as with Gentian Violet for its antibacterial properties. It looks like I'm just going to have to go ahead and do some experiments myself with that, because nobody else has used stains for medicine in the living cell cultures.

Here are my new babies in the bone soup, staying warm in the incubator.