August 28: Count twice treat once

First round of experiments. Fingers crossed that I:

#1: Plates the right number of cells

#2: Managed not to contaminate them

#3: Calculated the right concentration of drugs 

#4: Treated for the right length of time

So many ways for the Clinical Fellow Factor to duck my results 

August 26: FOILED

So today Leah and I spent an hour working out my first round of treatments. If we want to finish them at the same time they need to be staggered 12, 18

And 24 hours apart. They also need to be read when she's there to help use the machine then back track to count/plate the cells 48hrs before treatment. After all that work we settle on split today, treat

At noon, midnight and 0600 on Fri-Sat. Just to realzie oops it's 72 hours of treatments for the MTT and we didn't plate enough cells for the western. 

August 24: bug juice

After 2 contaminations in as many weeks, I found out we were supposed to be using pen-strep (1% penicillin-streptomycin) antibiotics in my media. Communication fail. 

August 17: gross contamination

Sacrificing my first cells in bleach 

August 14: My babies

It's my 32nd birthday and these are my children: fresly split and tucked in for the weekend!

August 13: Making omelets

Working on a combined project with Boston Children's. We are looking at the effect of omega-3 on prolonging duration of fertility in aged mice. We get between 0-9 eggs per retrieval. As I am blinded to the mouse's diet, I pretend the copious eggs are treatment and zero eggs are not (helps me stay positive). 

August 6: Adventures in peppers

The best and worst thing about being a fellow is teaching the residents... Maybe next time don't bend the resectascope

August 5: Don't throw your cells away!

This sounds rather obvious but once done splitting the extras aren't trash their experiments (it's like when you clamp cut tie but cut on the wrong side of the clamp). 

August 1: Fellowship complete: 3%

So I am 4 weeks old, so what have I learned thus far?
Then: I thought basic research worked like this: you get cells (in this case provided by the NIH), you run experiments (test for receptors, DNA/RNA etc) you publish data.

Now: you get a sample (NIH or from your patient), you grow them (in the right media, on the right plates, careful not to kill them), infect them with retro/lenti virus (after figuring out how much virus you need), select for the ones who grow the right genes (test by looking for GFP positive and then run western to confirm), split them make them grow more, freeze some (in case you kill them), grow and split again.  Use the same that you dont need to run experiments (in triplicate).  Keep growing them (increasing number passage) so you can do more experiments: proliferation assays, PCR or westerns.

Wow.  That's way more exhausting.
And that's without the CFF.  This is the term the ONC team coined for  the Clinical Fellow Factor (ie when I fuck it all up and throw cells away instead of doing PCR on them).

July 30: Utah SREI Retreat

July 21: We're #1