Hello Professors and Senior Researchers,
I am currently working on primary neuron culture.
I would like to share an issue I have encountered:
These primary neurons are very important samples for my current research 😢. I would greatly appreciate any insights or advice from senior researchers who have already mastered this technique. Any additional tips or responses would also be deeply appreciated. Thank you in advance!
You have axon fragments as debri. You may try gradient centrifuge to get rid of them and / or give the plates more washes after cell attachment. It seems you may have some contamination (small black dots) as well. Your cells look like neurons but to be sure you need to see them extend neurites. The arrow pointed cell is hard to identify.
Dear Seohyun:
Gurkan has given solid advice - make sure your cells are free from debris and you are super gentle with your trituration steps.
Also, you can verify:
1) You need a TC-treated culture dish or plasticware
2) You need a good coating of PDL or PLO, followed with a proper rinse
3) Try an alternative coating that doesn't degrade like dPGA
Source: I work at DendroTEK Biosciences, and our main product, dPGA is designed especially to stop clumping and peeling of neurons (primary and iPSC-derived) in cell culture.
www.dendrotek.ca