Previously, to perform a rheological study on the collagen matrix that we tried to make we used 2000uL of Collagen, 100uL of 10X PBS, and 13uL of NaOH. It formed the gel. For performing a better study we increase the total volume of collagen to 3000uL and added 150uL of 10X PBS along with 19uL of NaOH. The gel didn't form.
We used the same amount of incubation time and temperatures. The only difference is that we have used 24 well plate for the first one and tube with a lid for the second one. As it is tube, I had to mix it for longer time but gently.
I am not able to figure out the issue here. Any ideas, please? Thanks
Hello Haneesha Vishwa Sai
Collagen polymerization goes through two phases namely, nucleation phase in which molecular assembly takes place, and a rapid growth phase in which cross-linking takes place. This polymerization process depends upon several factors like surface area to volume ratio, volume and material used. The large surface area to volume ratio leads to more rapid temperature changes and faster termination of the nucleation phase (1st phase of collagen polymerization).
Also, the type of material used like in your case such as the 24-well plate in the first case and the tube that was used in the second case. Were they of the same material? 24-well plate is made of polystyrene which is a very good insulator because heat energy cannot flow through it.
So, the differences present between the 24-well plate and the tube could be the possible reason for the collagen gel not forming in the tube.
Best Wishes.
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Cell Biology
- Culture Cells
- Cell Line