@@chankingming8964 Great to hear from you and thank you for responding to my question! Cancer cell environment can be really acidic, I hope I can find papers about the stability of alamar blue and related methods. DMSO sounds good!
Thank you for your lecture! could you help to explain the MTT VS Alamer blue part. you said Alamer shows better result because copper sulphate has strong reductive power, so MTT approach may not be able to counteract the effects of the copper ions. i am so confused about this, so basically, live cells can produce a reductive items which can reduce MTT to from a purple product. copper sulphate has strong reductive power as well, so it should reduce MTT to from a purple product, that is why the cells all died out shown on the graph. (but actually there should still have cells alive just because MTT reduced by copper sulphate rather than the real reductive chemicals metabolised by live cells right?) so i am wondering, Alamer blue show better result, does it mean the Alamer blue is not sensitive to reduced by copper sulphate, so Alamer blue still get reduced by real reductive chemicals metabolised by live cells, namely NADPH or NADH. if it is true then why? and also for the reducing agents for MTT, are they NADPH or NADH as well? Thank you very much if you can help me !
Another question is , i noticed that when they do the Alamer Blue test: they add cell and copper sulfate first (4 h), then add Alamer Blue for 2h (for 6 hours group) and 20h (for the 24 hours group). But for the MTT test: they add cell and copper sulfate first (4 h) then add MTT for 2h for 6 hours group. but for the 24 hours group, they left cell and copper sulfate for 22h and then add MTT just for 2 hours. why they dont use the same time method as Alamer Blue test? is that the same reason that ALamer blue is not sensitive to be influenced by the copper sulfate, so we can add it earlier? thank you!
Exactly that alamer blue is more sensitive than MTT. MTT uses absorbance to detect the colour changes which could be affected by other situations like precipitation or colour from cells or the cultures
Exactly that alamer blue is more sensitive than MTT. MTT uses absorbance to detect the colour changes which could be affected by other situations like precipitation or colour from cells or the cultures
Exactly that alamer blue is more sensitive than MTT. MTT uses absorbance to detect the colour changes which could be affected by other situations like precipitation or colour from cells or the cultures
Exactly that alamer blue is more sensitive than MTT. MTT uses absorbance to detect the colour changes which could be affected by other situations like precipitation or colour from cells or the cultures
Exactly that alamer blue is more sensitive than MTT. MTT uses absorbance to detect the colour changes which could be affected by other situations like precipitation or colour from cells or the cultures.
Super relevant and easy to understand! Thank you so much sir! 🥰💕 More power!
Indeed this is useful
Thanks for watching!
Super well made video! Thank you!
Thanks for your complement
Great Presentation!
Thank you so much. Wonderful
Clear and well-explained! I wonder if these techniques will work in my experiment using a chemical to inhibit LDHA in tumors, especially alamar blue!
Why not? If the chemical is soluble in the culture medium well. If not you need to use an organic solvent such as DMSO to do the experiment.
@@chankingming8964 Great to hear from you and thank you for responding to my question! Cancer cell environment can be really acidic, I hope I can find papers about the stability of alamar blue and related methods. DMSO sounds good!
Very informative!
- Could you expand on how to calculate the LD50 value?
-What do you think of the Celltox green assay?
I use PRISM Graphpad SOFTWARE to calculate the LC value! Celltox green assay is fine
Hi, could you give examples of assays of membrane testing? Thank you in advance
Yes, LDH assay can somehow shows the leakage from membrane
Thank you for your lecture! could you help to explain the MTT VS Alamer blue part. you said Alamer shows better result because copper sulphate has strong reductive power, so MTT approach may not be able to counteract the effects of the copper ions. i am so confused about this, so basically, live cells can produce a reductive items which can reduce MTT to from a purple product. copper sulphate has strong reductive power as well, so it should reduce MTT to from a purple product, that is why the cells all died out shown on the graph. (but actually there should still have cells alive just because MTT reduced by copper sulphate rather than the real reductive chemicals metabolised by live cells right?) so i am wondering, Alamer blue show better result, does it mean the Alamer blue is not sensitive to reduced by copper sulphate, so Alamer blue still get reduced by real reductive chemicals metabolised by live cells, namely NADPH or NADH. if it is true then why? and also for the reducing agents for MTT, are they NADPH or NADH as well? Thank you very much if you can help me !
Thank u so much Sir... :)
Dear sir,where can i find this lecture as a PPTs ?
Another question is , i noticed that when they do the Alamer Blue test: they add cell and copper sulfate first (4 h), then add Alamer Blue for 2h (for 6 hours group) and 20h (for the 24 hours group).
But for the MTT test: they add cell and copper sulfate first (4 h) then add MTT for 2h for 6 hours group. but for the 24 hours group, they left cell and copper sulfate for 22h and then add MTT just for 2 hours. why they dont use the same time method as Alamer Blue test? is that the same reason that ALamer blue is not sensitive to be influenced by the copper sulfate, so we can add it earlier? thank you!
Exactly that alamer blue is more sensitive than MTT. MTT uses absorbance to detect the colour changes which could be affected by other situations like precipitation or colour from cells or the cultures
Exactly that alamer blue is more sensitive than MTT. MTT uses absorbance to detect the colour changes which could be affected by other situations like precipitation or colour from cells or the cultures
Exactly that alamer blue is more sensitive than MTT. MTT uses absorbance to detect the colour changes which could be affected by other situations like precipitation or colour from cells or the cultures
Exactly that alamer blue is more sensitive than MTT. MTT uses absorbance to detect the colour changes which could be affected by other situations like precipitation or colour from cells or the cultures
Exactly that alamer blue is more sensitive than MTT. MTT uses absorbance to detect the colour changes which could be affected by other situations like precipitation or colour from cells or the cultures.