This depends on 3 things 1) Resolution of your microscope 2) Magnitude of your pores 3) Clear images where you can see all the porous structures one by one

If you can see the clear images and pores together on the nanofibers obtained from microscope you can simply use the scale bar for all settings. But 50 micron scale bar is not promising. http://pubs.rsc.org/en/content/articlelanding/2016/ta/c5ta09166g#!divAbstract In this link you can find an article where you can see the real pores in Fgure 1c obtained by TEM. Therefore your light microscope images possibly dont have enough resolution to see such a detailed information.

For porosity measurement you can go for liquid displacement method rather than image analysis.

Dear Muna

You can install the "DiameterJ" plugin. It calculates pores, fiber diameter and orientation.

If you spin PLA in CHCl3 you might have fibers big enough for nanoCT scanners. Then you have the real scaffold pore sizes rather then 2D ones.

Sucess

Marc

Dear Mamatha M Pillai , is liquid displacement method for porosity measurement an ASTM standard method?

Dear Esrat Jahan,

Total porosity is the overall pore space present in a 3D scaffold. Image J can be used for measuremnt of pore size . Liquid displacement method is one of the simplest methods which is generally used to understand the porosity of a scaffold. There are other standard methods to analyse porosity of scaffolds such as mercury porsoimetry or BET depending on the nature of your scaffolds. You can refer to ISO/ ASTM standards methods for the same.

(https://www.iso.org/obp/ui/#iso:std:iso:15901:-1:ed-2:v1:en)

All the best

Thankig you,

Mamatha

Check out my record for determining fiber's diameter and the total area of pores in an SEM image via ImageJ software.

https://youtu.be/uKTewLu66oI