You will be provided with a DOT kit including everything you need to measure three human mockup samples (HUMOs) in two replicates. After performing the assay, you will upload your data to our portal, where all results will be combined into a population study that will be published in a high impact journal. Participants who successfully complete the measurements will have the opportunity to be listed as co-authors on the final publication.
The HUMOs provided in the kit come in three different phenotypes. These phenotypes are defined by the average number of target cells (HaCaT cell line, EpCAM+/EGFR+) that are dispersed amongst a much larger number of background cells. They are:
Negative HUMO: 0 targets/100,000 cells
Low HUMO: 1 target/100,000 cells
High HUMO: 20 targets/100,000 cells
*Note that these numbers are just averages and that the actual counts will be randomly distributed.
Your kit will contain one of each of the three HUMO phenotypes. You will analyze each phenotype in a single DOT reaction for which you will make 8 replicate measurements. Here is what you can expect to see: Negative HUMOs will return no hits and only a baseline signal for all 8 replicates. Low HUMOs will contain no hits in roughly half of DOT reactions, and half will contain 1 target or more in the 8 replicates. High HUMOs will contain hits in 5 replicates or more out of 8 for 90% of reactions.
For individual participants, the differences between HUMOs will appear statistically insignificant due to the small sample size. Most Low HUMO samples will contain no targets and appear indistinguishable from the negative phenotype. However, when the data from all participants is analyzed collectively, together we will be able to precisely discriminate between the three phenotypes. This is only possible because DOT technology has a false positive rate of zero and a well defined signal to target ratio. To provide a benchmark, it has taken over 100 hours to perform this analysis in our quality controlled laboratory at Sampling Human. Through decentralization and parallel processing, your collective data will show that the same results are reproducible in just 15 minutes of hands-on time.
Pictured In the figure are expected distributions of targets in each DOT reaction for the three HUMO samples. Each DOT reaction is sub-sampled from a total of 107 background cells with the number of target cells being phenotype-dependent (no targets in the Negative HUMO, and at frequencies of 2x10-5 and 10-6 for High and Low HUMOs respectively). The total cells analyzed by a single DOT reaction is 5.6x105.