For MTT assay, NRU assay and genotoxicity assay which microscope should I chose inverted microscope, inverted fluoroscence microscope or both seperately inverted microscope and fluoroscence microscope...
Dear Roma Lal
I am not sure if I understood your exact question.
As per my knowledge, if you perform MTT assay then you need a spectrophotometer to measure the absorbance at OD at 500-600nm.
So I do not follow your question about microscopy for this assay because you would actually need a spectrophotometer for MTT readout.
There has been a previous discussion thread on RG regarding MTT data analysis which you can also check out https://www.researchgate.net/post/How_do_I_calculate_cell_viability_in_MTT_assay
Also for NRU assay you need a colorimetric multi-plate reader.
Hope this helps you to figure out your experimental plans.
Separate from the assay measurement, I can suggest you to go through this webpage - https://www.keyence.com/ss/products/microscope/bz-x/study/principle/type.jsp
this might help you to get a basic idea of different types of microscope. Inverted microscope is the category based on the structure of microscope whereas fluorescence microscope is categorisation based on optical function.
Best wishes
Thanku Shambhabi Chatterjee mam for your valuable response....We have UV/Vis spectrophotometer....If that can be used for NRU assay.......Further for cell number count before MTT or to check contamination in culture is it required to have inverted microscope......I m performing it first time....And we do not have either inverted or fluoroscence microscope.....So i have to purchase it before that if it necessary.......If we can perform Cell culture without these sophisticated instruments....
Dear Roma Lal
Most UV/Vis spectrophotometer should work for these assays. However, it is best to double check if the spectrophotometer in your lab can detect OD between 500-700nm. This is the range that will work for both MTT and NRU assay.
If you need to check for contamination in cell culture then a normal inverted microscope is sufficient.
If you plan to stain your cells with DAPI or Hoechst to count nuclei only then you would need to purchase a fluorescence microscope. However, as far as I know you cannot use the same wells for MTT assay and cell counting. If you want to normalize the data with cell count then you should perform the entre experiment in 2 sets - one set to be used for MTT assay and the second set you will need to fix when you start the MTT test. Later on you can stain your fixed cells and count cell numbers.
So the instruments you will need to purchase will depend on the experimental steps that you will perform.
If you are doing it for the first time then I recommend to watch these youtube video - https://www.youtube.com/watch?v=hhAs1z4NeUE
https://www.youtube.com/watch?v=yPUUhJP2vwI&t=147s
This might give you a better idea about all instruments and reagents you will need to arrange before you can really perform this experiment.
Good luck !
- Badges
- Science method
- Similar topics
- Biological Science
- Histology
- Staining