Biology Forum Cell Biology Measuring OD

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    • #14258
      pab94055
      Participant

      Hello,

      I want to measure the OD of various bacteria. the plate reader I have can only read at OD595. Will that be ok if its not exactly OD600 as the literature suggest?

      Thanks

    • #102708
      JackBean
      Participant

      I would try to measure full spectrum first, but I think, it should be OK (although you will have smaller optical density!).

    • #102716
      canalon
      Participant

      Actually the wave length does not matter at all.
      When you are measuring cell density with a spectrophotometer, you are not really measuring the absorption of light, but its scattering by the particles (cells). And it is the same at any wavelength. The only thing that you might want to recheck is the equivalence OD=number of cells, as the values ate different wavelength will vary with many parameter (but that should be true even at 600nm, because the geometry of the different species of bacteria will influence the equivalence between OD and cell numbers). Yet honestly I am ready to bet that there will be no difference between the 2 measurements.

    • #102727
      jwalin
      Participant

      hmm that is an interesting point…
      hmm wait but isnt OD equivalent to absorbance?

    • #102759
      canalon
      Participant
      quote jwalin:

      hmm that is an interesting point…
      hmm wait but isnt OD equivalent to absorbance?

      No they are not. The bacteria are not absorbing the light. You are truly measuring absorbance when you measure concentration of molecules in solutions, like DNA (260nm) or proteins or pigment. But for particles like bacteria, it is just the scattering of the light (% transmission) that is measured, so the wave length is not really important as long as the blank is correct.

    • #103011
      JackBean
      Participant

      canalon: it’s not exactly the same, I think up to ~300 (maybe 400), there is no OD, but then it grows up.

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