I really enjoy benchfly videos :) Thank you so much! Though for this particular video I would like to note that the calculated surface areas are different. 10 cm dish will be pi*diameter^2/4, so 78 cm2 and 15 cm dish will be 176 cm2, so the ratio will be 1: 2.25, so to get 1:6 final dilution, you need 1: 2.67 of your cells from dish10cm -> so you need to take 10/2.67=3.8 ml cells to the dish15cm and not 5 ml.
So to simplify this video, you could use the equation: (Original confluence)(Original area)(Unknown Muliple of Cell Suspension)=(Target Confluence)(Target Area) Fill in the blanks. I think that works.... Let me know if it's wrong.
I had to watch this video twice to understand. I'm happy that you taught this slowly and clearly. The part where it got me a little bit confused since from 55cm2 to 148cm2 gives a 1 to 2.69... so I guess the numbers are rounded whole (2.69 -> 3)? Well anyway, the math seemed to work after that. However, the other confusing part is that do I absolutely have to make my cell suspension 10 mL? I mean, so long as a I make an appropriate suspension and be able to pipette exactly half of that is ok?
~ 10cm dish 15cm dish To Split 6 to 1 Surface area 1 to 2.25 3.14 X 5 X 5 to 3.14 X 7.5 X 7.5 (Assumption) This means I would have to transfer (2.25/6 =) 0.375 times the cells to the 15cm flask after adjusting for the difference in surface areas. Now if I made up the final volume in the 10cm flask to 10ml, I would transfer (10 X 0.375 =) 3.75ml of its evenly suspended contents to the 15 cm dish to achieve the desired result.
Pls can anyone explain how the person in the video calculated the surface area? As i know the simplest equation is pi*r*r. But according to this equation the surface area seems to be different compared to the person in video. So I do wounder whether the person used different equation?
This is wrong on so many levels. The initial size of the dish doesn't matter. Confluence is a subjective take on density. You need a hemocytometer to get the actual density and go from there.
I really enjoy benchfly videos :) Thank you so much!
Though for this particular video I would like to note that the calculated surface areas are different. 10 cm dish will be pi*diameter^2/4, so 78 cm2 and 15 cm dish will be 176 cm2, so the ratio will be 1: 2.25, so to get 1:6 final dilution, you need 1: 2.67 of your cells from dish10cm -> so you need to take 10/2.67=3.8 ml cells to the dish15cm and not 5 ml.
Samantics
Thank you 😊🙏
Remember adherent cell splitting is based on surface area (SA), suspension cells are not :)
So to simplify this video, you could use the equation:
(Original confluence)(Original area)(Unknown Muliple of Cell Suspension)=(Target Confluence)(Target Area)
Fill in the blanks.
I think that works.... Let me know if it's wrong.
I had to watch this video twice to understand. I'm happy that you taught this slowly and clearly. The part where it got me a little bit confused since from 55cm2 to 148cm2 gives a 1 to 2.69... so I guess the numbers are rounded whole (2.69 -> 3)? Well anyway, the math seemed to work after that. However, the other confusing part is that do I absolutely have to make my cell suspension 10 mL? I mean, so long as a I make an appropriate suspension and be able to pipette exactly half of that is ok?
~ 10cm dish 15cm dish
To Split 6 to 1
Surface area 1 to 2.25 3.14 X 5 X 5 to 3.14 X 7.5 X 7.5 (Assumption)
This means I would have to transfer (2.25/6 =) 0.375 times the cells to the 15cm flask after adjusting for the difference in surface areas.
Now if I made up the final volume in the 10cm flask to 10ml, I would transfer (10 X 0.375 =) 3.75ml of its evenly suspended contents to the 15 cm dish to achieve the desired result.
Excellent....I was going nuts in my lab today ha ha....this helped... thanks!!
If the video editor didn't realize that the tutor has been using color marker. Trying to make fancy doesn't help the view at all.
You basically need to take a 6th of the cells and multiply it by the scaling factor
Pls can anyone explain how the person in the video calculated the surface area? As i know the simplest equation is pi*r*r. But according to this equation the surface area seems to be different compared to the person in video. So I do wounder whether the person used different equation?
No, he simply wrote down casual areas.
Surface area's of dishes or flasks can be found by googling the info. They usually have tables that correspond to flask size and surface area.
This is wrong on so many levels. The initial size of the dish doesn't matter. Confluence is a subjective take on density. You need a hemocytometer to get the actual density and go from there.
it's really not so complicated