How can I quantify the extent of filamentation in C. neoformans?

11 November 2017 3 1K Report

I tried 2 methods:

1) Measuring the radial distance between point-of-origin (on the mating spot) of the hyphae to the tip. The shortcoming of this method, I find, is that it is prone to error on my part, especially when the filaments are slightly older and thus, more tangled and branched.

2) Converting the image to greyscale and measuring the intensity of the image. The idea behind this is that increased filamentation = decreased image intensity. However, it isn't very accurate as far as invasive growth in 3 dimensions is concerned, especially closer to the point of origin.

Does anyone know of a better method? Or can someone suggest an improvement on these 2?

Details that may be of use:

The fungus filaments only on a few particular types of solid media.

I image the periphery of my spots under 10x and stitch together the images of larger patches of filaments.

Neeran Jasim

you can see following link :

Article Blue light negatively regulates the sexual filamentation via...

it help you

Jake Winiski

Hello Bornika,

Dr. David Barry has published a number of papers regarding automated hyphal network analysis via the ImageJ plugin AnaMorf that I think you may find helpful.

Article Automated Analysis of Filamentous Microbial Morphology with AnaMorf

Article Nitrocellulose as a General Tool for Fungal Slide Mounts

Article Morphological quantification of filamentous fungal developme...

Download for AnaMorf plugin: https://bitbucket.org/djpbarry/anamorf/wiki/Home

Jawad K. Abood Al-Janabi

Hello Bornika Roy,

To examine the microscopic morphology of filamentous fungi I advise you to read Dave Barry's publications e.g. Image Analyst, Advanced Light Microscopy, under the title “Morphological Quantification of Filamentous Microbes”. At “https://bitbucket.org/djpbarry/anamorf/wiki/Home

Also check the following references:

1- Barry DJ, Chan C, Williams GA. 2009. Morphological quantification of filamentous fungal development using membrane immobilization and automatic image analysis. J Ind Microbiol Biotechnol 36:787–800.

2- Barry DJ. 2013. Quantifying the branching frequency of virtual filamentous microbes using fractal analysis. Biotechnol Bioeng 110:437–447.

Good luck

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