Is there any simple comprehensive way to calculate scale bar in microscopy images? Please mention in detail. Many thanks in advance.
If you do not have the scale slide the easiest method is to take something with known dimension given by the company who produce it, like thin gold wire or AFM cantilever, etc. Do the image at the same magnification and then scale the size of it to your image.
Thanks for your answer. I have dimension in pixels. Can we calculate scale bar from dimensions of image like width etc?
No. If you don't have any indication about the size of your object or the precise size of the field of vision, you cannot conclude the size of your pixels.
Hello Meha,
To be very specific, you can calculate the scale bar in any image by defining the magnifications. We use Nikon Eclipse TS100 and to put a scale bar on your image which is in pixels, you need to go through these simple steps.
Open the image-----Right click om image-----------Image properties----- go to Image fields/caliberation--------In first line where Uncaliberated will be written---Click on the button on the right end ther---You will get magnification option like 10X (0.69um) and so on---------Define your magnification and click-----Apply.
Now your image is ready to put scale bar. You click on the scale bar and it will provide the scale bar in um. You can edit the scale bar color, width and length depending on your choice by going to properties in scale bar. Once scale bar is put, click---Burn the scale and image will be saved with the same..
Hope it helps.
Thanks Ajay. That was comprehensive. We use Nikon TE2000E. NIS element is the software we use. So do you suggest this calculation on NIS element or shall I use image J for this purpose? Many thanks once again.
Hello Meha,
I do use NIS element only and this procedure is followed in that software only..It will work pretty well. Don't worry.
Cheers!
Hi Meha,
Just remember that this is an uncalibrated measurement so they are only "relative units"; at higher magnifications there will be more and more error in your measurements. To make sure that the scale bar is correct you should measure something with known dimensions like Katarzyna suggested, another good thing to measure is the distance between lines on a counting chamber.
As Katarzyna aslo said, most companies will make specific slides to calibrate the digital measurements.
Good luck!
Nick
@ Nick and Meha,
dear that is not an uncaliberated measurement. If you will go into details of the procedure, the scale is calibrated with the help of the standard scales which the company engineer will have on his calibration glass slides. The distance between two points is known on that slide so that distance is set as 10um or 20um depending on how may points are selected. The scale is then assigned values at different magnifications. That's a one man job and one can easily do calibration if you save fotos of calibration scale also.
So in nutshell, that is 100% caliberated and reliable.
Cheers!
Please let me know how we can calculate scale bar of 10 X magnification in image of microscope ?
I am using a Labomed Compound microscope to study the histology of layers of GIT and neurons. I have taken pictures on the magnification of 400X (10X eyepiece x 40X objective lens) and 1000X (10X eyepiece x 100X objective lens). How to calculate the scale bar for these images taken on these magnifications? How to add the scale bars in the images taken on this magnification?
Kindly help me.
I have clicked pics in leica research microscope but settings are not changing while clicking. Values are same for every magnification . How to rectify the error on already clicked images and settings
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