To perform this evaluation I used a quite easy and cheap method, I got negative impressions applying nail polish as Phillip Glombik said, then after some seconds I recovered those polish layers with adhesive clear tape and visualized them in an inverted microscope. I hope this helps you.

I have also used the nail polish method with an inverted microscope. 

Article Nocturnal transpiration in riparian Tamarix thickets authent...

There are several ways to estimate stomatal density.  1) Take a digital image of the microscope field, if it has a scale for reference. Then you can use image processing software. 2) if microscope has a grid, you can count the number in each square.

its important to know the scale of magnification for accurate density estimates.

I agree with the nail varnish method, but you can also make impressions with dental putty first, then paint those with nail varnish, peel them off and use the microscope to see. This way you get to keep intact impressions and can redo them easily.

Thank you all for valuable suggestions, I work with Arabidopsis thaliana

Hi James,

We have recently published a boo chapter on guard cell isolation- very extensive and easy to follow.

Zhu, M., Jeon, B. W., Geng, S., Yu, Y., Balmant, K., Chen, S., & Assmann, S. M. (2015). Preparation of Epidermal Peels and Guard Cell Protoplasts for Cellular, Electrophysiological, and-Omics Assays of Guard Cell Function. Methods in molecular biology (Clifton, NJ), 1363, 89-121. Link:http://download.springer.com/static/pdf/118/chp%253A10.1007%252F978-1-4939-3115-6_9.pdf?originUrl=http%3A%2F%2Flink.springer.com%2Fprotocol%2F10.1007%2F978-1-4939-3115-6_9&token2=exp=1449164544~acl=%2Fstatic%2Fpdf%2F118%2Fchp%25253A10.1007%25252F978-1-4939-3115-6_9.pdf%3ForiginUrl%3Dhttp%253A%252F%252Flink.springer.com%252Fprotocol%252F10.1007%252F978-1-4939-3115-6_9*~hmac=ea19cba29c7d5406f5b601ed0123fd23760d3843578a182e8f48b1170fe84d69

Thanks,

Biswa

Hi, is there non-destruction method (s) of measuring stomatal density in the maize leaf? please suggest.

Try this tool that I and a few collaborators created. It's called StomataCounter and you can use it at http://stomata.science. It's a fully automated method based on a deep learning network trained on about 700 species and several thousand cuticle images.

Thank you, Mr. Karl Fetter, to give me an excellent resource.

Karl Fetter Hi . I am trying to run StomataCounter on a sample image here and it's been "Processing" for almost 40 minutes. Did I do something wrong? I tried deleting and uploading it again, but it's still the same thing. Any advice?

I'd also recommend the Multi-Point tool on ImageJ, although it seems very laborious!

I used it to count bacterial and fungal colonies on 15mm petri dishes; we got up to 1000 colonies per plate and it was not that terrible as it sounds, because it's really quick to do! haha

There are different methods, some people use nail paint cover to elaborate the content of stomata and clear for count.

Thank you!

I have written few scripts to count the stomata automatically regardless of the image quality.

https://github.com/sivkri/Count-stomata-and-pavement-cells