Slide Culture Preparation: Introduction, Requirement, Procedure, Uses and Disadvantages

achaurasiya

5 years ago

Slide Culture Preparation: Slide culture preparation or technique method is also called microculture method. Slide culture preparation is useful in mycology due to following points of views- in order to accurately identify many fungi it is essential to observe the precise arrangement of the copnidiophores and also to know the nature of spore production Riddel's used a simple method of slide culturing but it permits fungi to be studied virtually in situ with as little disturbance as possible. But here, we use a simple modification of his method using a single agar plate as described below.

Introduction of Slide Culture Preparation

Slide culture preparation or technique method is also called the microculture method. Slide culture preparation is useful in mycology due to the following points of view-

in order to accurately identify many fungi
it is essential to observe the precise arrangement of the conidiophores
and also to know the nature of spore production

Riddel’s used a simple method of slide culturing but it permits fungi to be studied virtually in situ with as little disturbance as possible. But here, we use a simple modification of his method using a single agar plate as described below.

Requirements for Slide culture preparation

Sterile Petri dish
Spirit swabs
Sterile Scalpel blade
Filter paper (sterilized)
Media- 1 / 1 cm units ( normally PDA, however, some fastidious fungi may require harsher media to induce sporulation like CMA, Czapek-Dox agar)
Stick or glass rod
Sterile slides
Distilled water
Specimen ( test fungal growth )
LPCB stain
Nail Polish
Cover-slips
95% ethanol

The procedure of Slide culture preparation

All of the processes should be done with aseptic precaution.
Cut 4 pieces of media measuring 1/1 cm (according to the size of the slide)
Inoculate the four sides of the agar block with spores or mycelial fragments of the fungus to be grown and cover with a flamed coverslip on top after inoculation.
Incubate the plate at 26°C until growth and sporulation have occurred.
If necessary, put the filter (wet) on the button of the Petri dish. Prevent the dryness of media but we just added water.
Remove the coverslip from the agar block and apply a drop of 95% alcohol as a wetting agent.
Gently lower the coverslip onto a small drop of LPCB on a clean glass slide.
The slide can be left overnight to dry and later sealed with fingernail polish
When sealing with nail polish use a coat of clear polish followed by one coat of red-colored polish.
Focus at 10X objective and finally observe at 40X objective under the microscope to observe fungal structures ( hyphae, spores, etc.).

Advantage of Slide culture preparation

Preserve the intact fructifying fungus for solidarity.
Attachment of filamentous fungus.
Arrangement and types of spores.

The disadvantage of the Slide culture Technique

As the agar medium used for Slide culture contains a limited nutrient source, fungal pathogens will survive for a short time period.
Slide cultures preparation of slow-growing organisms suspected to be dimorphic pathogens such as H. capsulatum, B. dermititidis, C. immitis, P. brasiliensis, or S. schenckii should not be made because high-risk pathogens may cause laboratory-acquired infections.

Key Notes on Slide Culture Preparation

Make at least 2 blocks per culture.
Some authors suggest the size of the agar block should be 7×7 mm i.e. small enough to fit under a coverslip.
Fungi when grown in a nutrition deficient medium ( e.g. PDA, CMA) develop spores rapidly and adhere to the surface of the coverslip.