Try the slide culture technique -http://www.mycology.adelaide.edu.au/Laboratory_Methods/Microscopy_Techniques_and_Stains/slide.html

Two  methods you can use first slide culture you put cover slip on Fungi colony then take it with care and safety consideration use of mask and bio safety cabinet  put on glass slide put a drop methylene blue from slide margin you can watch directly  Mycelium structure with its spore under microscope in 10 .

Second you must take a sample of colony fix it by alcohol and then staining it  but may be you can watch a portion of Mycelium  and spore in fixed condition but in mycology safety issues must be consider seriously because of opportunistic fungi risk.  

For identification of fungi, its hyphae  that are filaments and make up all structures of multicellular fungi should visualize. Some hyphae have cross-walls called septa (the singular form of septa is septum) between cells, and others do not. A network of hyphae reaching through acres of soil or covering an orange peel, for example, is called a mycelium.Filamentous fungi grow radially in search of nutrients. It is important to

understand the growth zones as you look at growth on a solid nutrient agar plate. When describing a fungus or sub-culturing a fungus or inspecting a specimem

microscopically, you will need to be able to discern the fungus growth stages. The cells

grow apically from the hyphal tips.

 A slide culture is the best method for observing sporulation characteristics of most fungi for identification purposes, though it is still difficult to discern separate fungal .

You can also prepare a direct mount of fungi. This method is not as satisfactory as

slide cultures for fungi identification through sporulation patterns. Direct mounts

are, nevertheless, more quickly prepared and adequate for preliminary observations.

You can try this simple adhesive tape method:

http://mycology.adelaide.edu.au/Laboratory_Methods/Microscopy_Techniques_and_Stains/cellotape.html

http://thunderhouse4-yuri.blogspot.sk/2012/06/adhesive-tape-preparation-for.html

Be sure the culture is pure . With the help of inoculating needle tease some portion of growth of the fungus and place it on the slide. If the fungus is heavy sporing like Aspergillu or Penicillium wash with 70% ethanol., remove ethanol by blotting paper. Then keep a drop of Lacto-phenol cotton blue, spread the mycelium with needles , place a cover slip. Examine . Note morphology of hyphae, spore structures etc. Consult keys given in standard books on mycology and identify.

Whatever may be the method of preparation of slides or samples you have to have through knowledge about the morphology of the fungi you wanted to study. But you must note the following characteristics: (i) Colony morphology and colour in a particular medium.

(ii) Appearance and morphology of hyphae/mycelium

(iii) shape, size and nature of spores.

(iv) shape and size of spore bearing structure and number of spores in spore bearing structure.

Double cover glass method ( Kohlmeyer, Marine mycology)can be used for Permanent slides preparation . for common observation You can take small hyphae from culture with ste rile needle transfer it to a drop of lactophenol cotton blue and covering with overslip. which can be observed under microscope

There are various ways you can identify and differentiate the fungi. We can get many information from morphology of fungi and from its spore and mycelium characteristics. Please refer to the paper i have attached. Hope it will be helpful for you. 

Movies : slide culture technique for fungi

https://www.youtube.com/watch?v=FcW5tHrbAos

There many  methods you can used  to identify the fungi under microscope . Please show my  paper attached with these message .

I hope to help for you . 

In almost all cases, you need conidia and/or sexual spores in order to identify a fungus based on morphology. Personally, I would always back up a morphological identification with a sequence based method (ITS sequence) or a protein based approach (MALDI-TOF).

you have to search for conidia not only the hyphae. If there is no conidia you may need to reculture the fungs on another media

Culture the fungi on potato dextrose agar, which stimulate conidia growth,  in a slide culture.  Use lactophenol cotton blue to see microscopic morphology.

Ensure that after adding the lactophenol cotton blue on slide, you gently lift your mycelium onto the slide and have it neatly teased out for identification

When viewing with low power objectives you need not cover with cover slip for it to be clear, then apply cover slip onto the mounted sample and add a drop of oil immersion and view with x100 objective lens.

Fungi are identified by their morphology in culture. Fungi have mycelium and spores which are used in the identification. Therefore you have to search for mycelium (hyphae), the spores, origin of the spores, asexual or sexual; and their structure and morphology.  So you have to see the morphology clearly.  Cut  small piece of fungal growth with an inoculating needle, place it on the slide with a drop of  lactophenol, KOH or water, tease the fungal growth with teasing needles so that hyphae spread sparsely, place a cover slip. If the fungal growth is heavy sporing wash with 70 per cent ethanol so that clumps are removed. Now  examine for hyphae, conidiophores (spore  bearing hyphae) spores.  Note down the charatcers of growth, hyphae spore bearing  hyphae and spore nature. Search the web using your data.. Visit nearby laboratory which deals with fungi. Calcutta School of Tropical Medicine is one such.

You have to look to the following characteristics:

1-   Fungal Morphology

2-   Fungal structures

3-   Reproduction in particular asexual spores: SPORANGIOSPORES and CONIDIA.

·       Sporangiospores:

v ENDOGENOUS - formed and contained WITHIN a  SPORANGIUM, ZOOSPORES (motile) and  PLANOSPORES (non-motile).

·       Conidia:

v EXOGENOUS - often formed at the tip of supporting hyphae called a CONIDIOPHORES, Two main types - THALLIC and BLASTIC

Also there are on line key for some plant pathogenic fungi e.g. see this link http://www.omafra.gov.on.ca/IPM/english/cucurbits/identification-keys/disease-seedling-discoloured.html

Good luck and best regards

Fungus can be identified easily by using Wet mount technique mostly but slide culture technique can be used for more clear morphology. You can little modify the method you are using for making slide by adding one or two drops of Lactophenol Cotton Blue first on slide, then  transfer  little growth of fungi in this drop and then tease this growth gently with the help of two needles to make the mycelium apart and give clear image in microscope.

Wishing you success

To identify fungi  under microscope the best technique is a slide culture, first from the primary plate/tube make slide culture of overnight to 48 hours. Then take a slide add a drop of lactophenol cotton blue, then carefully pick the cover slide from the slide culture and place on the slide that contains the lactophenol with out disturbing. Exam under microscope with high power objective. (note: don't disturb  the connection between  hyphal and conidia structure because it  is important for the identification). If you prepare the slide carefully you can easily observe the association  between hyphae and conidia which is very important and help for the identification.  You can also make another slide for additional preparation, pick the agar from the slide, discard it and add a drop of  lactophenol cotton blue on the slide, cover with cover slip (don't mix) and exam under microscope.

Without the appropriate understanding of classical mycology and its taxonomy, please understand application of the above techniques is unlikely to give you a reliable identification.  You're probably best advised to get the help of a trained mycologist.

Dear Mr 

After added  the mycelium  on slide , don't  move  the mycelium  quickly  because this moved was destroyed the mycelium. 

Pick few distinct hyphae with needle and place on clean glass slide. Add lithe lactophenol blue stain and allow to react for 2 to 3 minutes the drain excess stain and cover with a slip. Observe under microscope and compare the structure observed with diagrams/pictures/drawings of known fungi. You also need to combine this with the characteristic features such as colour shown on solid media like PDA, SDA, MEA, czapex dox agar.

I am also looking for the best and easy way of mounting fungi and algae but have not received any good suggestion.

Place a drop of ethanol on the slide, lift a few strands of the fungal hyphae with the aid of fine forceps and needle and place in it. Add few drops of cotton blue lactophenol onto the preparation.Gently lower a cover slip onto it avoiding air bubbles. If air is traped add a drop of the stain at one edge of the slide and it will flow in to drive it out .

Thank you.

1) before preparing of the slide you should examine the front and rear face of the culture.

2) Use an adhesive tape instead of the cover slide , is better.

3) You should depend on a mycology atlas , (picture key) .

when you observe microscopic examination of fungal culture 1st of all prepare a young culture of fungi then take a mycelial growth with the help of needle (not a inoculating loop) and spread the growth on slide then add a drop of 50% diluted cotton blue stain and cover with the help of cover slip is better

Hello shovon, don't use the coverslip during identification of fungus, because the mycelia will not appear very clear. Try this.

Identification of fungus under microscope is very easy with the helf of lactophenol cotton blue stain but we use ist of all upper mycellial growth with help of needle and tease(spread) the growth with the help of needle and drop a cotton blue stain

This method helps with the identification of Actinomycetes?

Hello every body , can any body help me how to identify fungus under microscope is there any reference i could found all fungus in it?

i find a perfect procedure for identification of fungi u i am sharing a web id here https://microbeonline.com/lactophenol-cotton-blue-lpcb-mounts-principle-staining-protocol/#:~:text=The%20lactophenol%20cotton%20blue%20(LPCB,The%20preparation%20has%20three%20components%3B&text=Lactic%20acid%3A%20It%20preserves%20fungal,in%20the%20fungal%20cell%20walls.

Dears, Shovon and Manal Al-musa,

Greetings,

You can not find all fungi in one reference. But you can depend on many different available references to identify the different fungi. This thing depends on the type of fungi that you want to isolate and identify.

Dear, Manal Al-Musa,

If you isolate the fungi from soil, seeds and rhizosphere you can depend on the references below:

1 - Compendium of soil fungi, by: Domsch et al. (1980)

2 - Pictorial atlas of soil and seed fungi: Morphologies of cultured fungi and Key to species, Second edition. By: Tsuneo Watanabe (2002)

Dear, Shovon,

According to my continuous working with isolation of fungi, I see it's very easy to see the mycelium of the fungus when I use lactophenol on the glass slide and coverslip. I take the mycelium from the Petri dish by using sterile needle and put it on the drop of lactophenol which put on the slide, then cover it with coverslip carefully and then see it under microscope with 10X firstly, then with 40X or 100X according to the size of sample.

If you want to isolate Aspergillus spp. or Penicillium spp., you can depend on the following references:

1- The genus Aspergillus, By: Rapper and Fennell(1973).

2 - The genus Penicillium and it's teleomorphic states Eupenicillium and Talaromyces, By Pitt, J. I. (1979)

When you want to isolate fungi from damaged or spoilage food and want to identify the isolated fungi, you can use the bellow references:

Fungi and food spoilage, By: Pitt and Hocking (1997)

Fungi and food spoilage, Third Edition By: Pitt and Hocking (2009)

Dear, Manal

If you deal with Fusarium spp., you can depend on the following references:

1 - The genus Fusarium. By: Booth (1971)

2 - The Fusarium laboratory manual. By: Leslie and Summerell (2006)

They are very useful for Fusarium identification.

To identify the fungi belonging to dematiaceae which contain the colored fungi you can return to the following references:

1 - Dematiaceous Hyphomycetes by: Ellis, 1970

2 - More Dematiaceous Hyphomycetes, By: Ellis, 1976.

These two references deal with the dematiaceous Hyphomycetec which produce colored conidia and conidiophores.

Hi Dr Shovon Lal Sarkar . To identify fungi under microscope the best technique is a slide culture, first from the primary plate/tube make slide culture of overnight to 48 hours. See the link: https://www.researchgate.net/post/What-is-the-best-method-for-the-identification-of-Fungus-under-microscope

LPCB and Slide Culture Technique

Technology of slide culture aside, you need to be trained in fungal identification based on morphology.

based on morphology, you prefer this book Larone's Medically Important Fungi: A Guide to Identification

Dear, Aref Wazwaz,

Yes, slide culture is a good technique, but it's not necessary for all fungi. Most of fungi can be tested easily under microscope by using lactophenol with cotton blue.

With regards.

Dear, Ved Prakash Mishra,

Greetings sir, If you have the reverence that you refer above (Larone's Medically Important Fungi: A Guide to Identification), please kindly, send it to me on this platform.

Fungus identification is a challenging process.

the books suggested above are all v good once you know the genus, but you have to get thst far.

1. Observation on the original plate is useful , colour, mode of growth etc, even differences in colour on top and bottom of the colony.

2. If the fungus is growing very vigorously , try a low nutrient medium such as tap water agar with i minute amount of malt

3. You really need to be able to see the spores in detail. How they are produced. Slide cultures or Tap water agar display this well. Traditional methods of I’d are based on spores not mycelium.

4. If it is a fungus causing disease in humans look at books and papers by Malcolm Richardson, emeritus professor of medical mycology at the uni of Manchester U.K. but remember that fungi are opportunistic pathogens and those not usually infecting humans can cause serious disease in predisposed humans. ( eg black fungus disease caused by Fusarium in Covid patients with diabetes and then treated with steroids.

I encounter challenges in trying to identify fungus on the stage of a microscope. can anyone help me the techniques to go about?

The attached files could be useful