On pregnancy day 18 we sacrificed one pregnant rat and we isolated the Hipothalami of the embryos (there were approximately 15 pups). All the Hypothalami were collected in one test tube. They were then further processed.
The isolated primary neurons were then seeded on a 6-well plate (3 Mio/well).
What is the N-number in this case?
Is it 1, since all the embryos derived from the same mother?
Or is it 6, since there are 6 wells?
The n = 1 with 6 technical replicates because you processed all of the embryos into a single tube before plating. If you had processed each embryo and plated the resulting cells separately your 'n' would be 15.
This will be equivalent to an experiment if you buy a cell line from a company (which majority of people conduct experiments--- People do not buy 10 cell lines of same type) and use the the same cell line in 15 wells, will that be n = 15 or =1 (since cell line is 1).
Hello Muriel Zanicolo
You should not have pooled the hypothalami because each embryo is different from the other.
In this case N is definitely not 6 because as John Hardy Lockhart mentioned, they would be technical replicates since you have pooled the hypothalami and from the pool you have plated 6 wells. So, N will be 1 not because it is derived from the same mother. Each embryo has its own identity. The primary culture you will obtain will be heterogeneous as they are from different embryos.
As John Hardy Lockhart mentioned, if you had processed each embryo separately and plated each embryo in 6 wells then N factor would be15. So you would have had 15 (6-well plates) with each plate dedicated for one embryo.
As Subhash C. Juneja mentioned, when 10 cell lines of the same type are used in 15 wells, the n factor will be 1 because the origin of the cell line is the same.
Best.
- Badges
- Science topic
- Similar topics
- Agriculture
- Horticulture
- Seeds