I have doubts about how many cells I can seed in different wells or plates. I would like to know some tips for determining this or to know a site that I can find this information.
Invitrogen has a pretty good table that should get you started I have attached the link below...
Invitrogen has a pretty good table that should get you started I have attached the link below...
Hi Priscila
Hidden in my drawer away from the prying fingers of PhD students is my little lab bible, the Roche Lab FAQs (http://www.roche-applied-science.com/PROD_INF/MANUALS/labfaqs/). Hit up your local Roche rep and get a copy - it's gold!
Al
Hi Priscila I'm doing MTTassay and seed 10000 K562 cell and PBMC per well in 96 well pate. but I seed 5000 cell per well of Hela, HEK293T or COS7 cell line in 96 well plate. becouse the size of these cells are different.
It really depend on what type of cell you are working with. How well they survive plating, how fast they proliferate. I work with primary cultures, and depending on what you culture, the plating number change.
not applicable to everyone, but for those with an iPhone/Android? there is a nifty app called "Daily Calcs" available from Life Technologies (Invitrogen) that has information regarding plating/media volumes/trypsin volumes, etc for various types of dishes, flasks and cluster plates. Its probably the same information Mark provided however.
One point i like to keep in mind when plating in very small vessels, e.g. 96 well plate's wells: do not make the layer of media too thick, it seems to interfere with aeration. I try not to exceed 100uL/well of 96 well plate.
Qiagen's RNeasy Handbook has a table (Table #4) similar to the one recommended above from Invitrogen, but I like it more, because it also has 48&96 well plates listed. However, it does not have media/trypsin volumes, just the HeLa cell numbers and surface area, so if you are looking for that information, stick to Invitrogen (personally, I prefer using much smaller volumes of trypsin, than those indicated there).
My gratitude to Alasdair for Roche manual reference!
Good luck with your experiments!
There are couple of handbooks which prescribe the cell density required to seed a flask or plate and it varies according the items (flask/petridishes/96 well plate/roller bottles etc) you use. I am attaching some pdf for your help
and my appreciation for Alasdair for Roche manual reference
All d best for your experiments
There are couple of handbooks which prescribe the cell density required to seed a flask or plate and it varies according the items (flask/petridishes/96 well plate/roller bottles etc) you use. I am attaching some pdf for your help
and my appreciation for Alasdair for Roche manual reference
All d best for your experiments
As i cannot attach two more files at one try therefore you would see two more post addressed by me
Seeding cells in different sized well?. Available from:
https://www.researchgate.net/post/Seeding_cells_in_different_sized_dishes_plates [accessed Aug 1, 2017].
Yousif
Does anybody works with human fibroblast passage 18-25?
what relation you use?
thanks
i was experiencing the same situation..i found all the answer is helpful...and i found this link work good too
https://www.thermofisher.com/us/en/home/references/gibco-cell-culture-basics/cell-culture-protocols/cell-culture-useful-numbers.html
Dear friends,
I came very late as part of this discussion but there was the same doubt to me. I developed a calculator in excel that will help you to estimate the number of dishes you need to subculture cells into if you know the confluency of your cells in old dish. if you have any questions, please ask me
Kind regards,
Ali Nasar
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