Posted by
Chloe van Oostende on
Feb 08, 2018; 4:19am
URL: http://imagej.273.s1.nabble.com/Splitting-images-Histograms-tp5019997p5020027.html
Hi,
First, you need to make sure you are in focus, or the best is to do a sum
projection of a z-stack to properly quantify the membrane localisation.
I find weird you get a 60.77 AU for one value. Did you draw the line in a
region where there is no proteins at the plasma membrane?
Otherwise, yes, 103.69 is your A.U, and yes, you need to have several cells
to measure...Usually 20 to 50 cells should be used.
For the distance, you can see that they wrote on the top of the graph. you
can calculate the natural logarithm from your x values if you want, but you
can also leave it in um as well ;)
Best,
Chloe
*Chloë van Oostende, PhD*
Chair,* Plant Science* session
Canadian Microscopy and Cytometry Symposium 2017
Facility manager- Senior Microscopy specialist
*Cell Biology and Image Acquisition Core Facility*
Faculty of Medicine
University of Ottawa
RGN 3171
451 Smyth Road
Ottawa, ON K1H 8M5
Office: 613-562-5800 ext: 8376
http://www.chloevanoostende.netCBIA core facility
<
http://www.med.uottawa.ca/research/corelabs/CoreLabs_CBIA/eng/>
2018-02-07 20:03 GMT-05:00 csadangi <
[hidden email]>:
> I tried what you suggested. First I drew a Vertical line and measured the
> plot profile. I exported it to an excel sheet and looked for the two
> highest
> values:
> X = 0.848; y = 141.41 - vertical
> X = 3.39; y = 106.44
>
> I repeated the same step by drawing a horizontal line
> X = 1.13; Y = 60,77 - Horizontal
> X = 3.10; Y = 106.14
>
> I averaged all the Y values and got 103.69. Can I mention this value as an
> arbitrary unit (A.U.) as they do it in the paper? How can I define an
> arbitrary unit? Also, do you suggest doing similar calculations for a
> number
> of other cells and then doing statistics? In the paper they say "The peak
> values of each line (2 values/line) were used to calculate peak fluorescent
> intensity of KCC2 at the membrane". So, can I say the peak fluorescent
> intensity of the membrane is 103.69 au
>
> Another doubt I have is that I cannot change the X-axis to log value as
> they
> show it in the paper. I changed the X-axis from Set Range option to display
> as log value, but it didn't change and it still shows in microns.
>
>
>
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