Microscopic examination of fungal smear

Jing Wu

Dermatology Department, Huangshi Central Hospital

wujingmed@163.com
Mycology concepts

Fungi belong to the biological kingdom and are an important

part of microbiology. There are many kinds, the number is up to tens o
f thousands
The big ones are like mushrooms,the small few micrometers c
an be seen with the help of a microscope.
Fungi include medical fungi, Medicinal fungi, industrial fungi,
edible fungi, Our main research field is medical fungi.
 Superficial fungus-cause various dermatophytes
Deep fungus-cause systemic fungal disease
Fungal allergy (asthma, allergic pneumonia, Allergic rhinitis,
allergic conjunctivitis, over Allergic dermatitis, etc.)
Fungal poisoning (fungal contamination or stored in food The
metabolites produced in the substance are mycotoxins, Can cause acut
e or chronic poisoning of people).
Aspergillus flavus and other fungi can produce carcinogens
♦ eukaryote
♦ cell wall

♦ no chloroplast
♦ saprobiotic or

parasitic
Definition of fungi

eukaryote
genetic material (chromosomes) enclosed within
a membrane-bound nucleus
cell wall
a rigid cell wall enclosing the whole cell
no chloroplast
saprobiotic or parasitic
The basic form of fungi: hyphae and spores

Hyphae: It is a fine tube with a smooth surface ,Consistent thickness and Bend
naturally.
The ends of mycelium are neat, Colorless, or colored (dark silk cells).
Separated, or not separated (Algae), Branched or unbranched,Multicellular
tubular structure.
There are 9 types of hyphae.
Simple hyphae and branched hyphae (mycelium)
Racket hyphae: the hyphae are enlarged at one end of the diaphragm, and
several sections are connected in a regular arrangement
Pectinate hyphae: One side of the mycelium is irregularly raised, like a long-
used comb, uneven
Spiral hyphae: Mycelium rotates regularly like a spring
Nodular hyphae: The hypha bends into clumps, like nodules
Articular hyphae: Articular spores arranged in clusters
Staghorn hyphae: The top of the hyphae has irregular protrusions (the
protrusions are not separated) like antlers
Pseudohyphae: The spores are arranged like hyphae, Constriction ring formed at
the junction of spore and spore
Arcuate hyphae: The hyphae are short and unbranched, arc-shaped, sausage-
shaped
Spores: It is the smallest unit of fungus reproduction and an important
basis for fungi classification and identification.

Spores: Is formed by a cell budding, When the bud grows to the size of the
mother cell, Break away from the mother cell or connect with the mother cell
and then regenerate buds, So repeatedly. Pseudohyphae. The width of the two
cells in the fungal filament septum is consistent, and the blastospores in fungi
are more common in Candida.
Thick-walled spores: It is a kind of dormant cell of fungus. The spores
are divided into free spores, terminal spores and interstitial spores.
Articular spores: It is formed by the breakage of articular hyphae,
short column or tube
Macroconidia: It is multicellular, spindle-shaped, rod-shaped or pear-shaped,
with smooth or thorny walls, and the number of separations is related to the
bacterial species. The morphology of macroconidia is the basic feature for
identifying fungi.
Microconidia: Unicellular, clustered or arranged in clusters, round or oval,
Separated from the conidiophore
Classification of Fungi

Dermatophyte
Yeast
Mould
Dermatophyte

Trichophyton:
Trichophyton rubrum
Trichophyton mentagrophytes
Flavus
Trichophyton violaceum
Trichophyton rubrum
Dermatophyte

Microsporum:
Microsporum canis
Microsporum gypsum
Microsporum rust
Epidermophyton:
Epidermophyton floccus
Yeast

Candida spp (Candida albicans, tropical, kerou, Smooth)

Malassezia
Mould

Aspergillus (Aspergillus flavus, Aspergillus fumigatus)

Penicillium (penicillium marneffei)
Fusarium (Fusarium moniliforme, Fusarium solani)
 Laboratory methods

 Microscopic examination of fungal smear

 Culture

 Pathological examination

 Wood’s light

 Reflexion confocal microscope, RCM

 Serological examination

 Biomolecular tests
Significance of direct inspection:

1. A positive microscopic examination indicates a fungal infection,

However, a negative microscopic examination cannot exclude the
diagnosis.
2. A positive microscopic examination can identify a few pathogenic
bacteria, such as tinea versicolor and yellow tinea Cryptococcus
neoformans, rhinosporum, etc.
Tinea pedis( 足癣）
Onychomycosis
( 甲真菌病 )
Pityriasis versicolor
( 花斑糠疹 )
Experimental procedure of fungal microscopy

Collect specimens

Specimen processing

Fungal microscopy

Audit Report
Principles of specimen collection

in lesions
Selection of general sk
Trunk Size New and old Location

Limbs
Small New Relatively i
nside
Soft and del
Scalp and hair
icate
Avoid fricti
Traumatic collection on
 Collect specimens

Secondary disinfection (be

fore and after collectio
Collecting knife
n）
Collection site 1. Prevent infection of the
collection site
First step ： disinfect 2. Prevent cross infection
3. Ensure the fungus cult
ure environment
Special parts Normal saline (mucosal area,
large trauma area)
The skin lesions are clearly d
Second step ： collection emarcated
Blurred boundaries of （ Groin, palms, soles of feet
skin lesions finger/toenails ）
Scalp and hair

1 、 Cut hair

2 、 Scab
Inner and outer spores, hyphae, pityrospor
es
3 、 Scales

4 、 Pus
 Traumatic collection

1 、 Scabies Hand stitches, palms, flexed wrists (corresp

onding to feet)
2 、 Vesicular papular lesions Inside skin lesions (blister type), secretio
ns (folliculitis)
3 、 Trichocystis Face (nose) and back (next to spine)
 Specimen processing

10%-15% KOHθ dissolves keratinocytes 压片厚

角质未溶

Cover glass, thin smear

If necessary, heat it on an alcohol lamp to pro

mote dissolution
 Fungal microscopy

欠佳
不合
格
Audit Report

The report of fungal microscopy is a clinical qualitative experiment

Specimen with only aseptic filaments of pityriasis spores under the

microscope should be cautious

Sputum specimens should be judged "whether the sputum specimen i

s qualified"

Oral specimens should exclude the interference of bacillus and actino

mycetes
 Microscope

10% KOH

blade

Alcohol lamp

glass slides
Suitable sampling position can increase the positive rate

 Tinea corporis:The edge of skin lesions

 Tinea capitis:Broken hair
 Onychomycosis:nail scraps
 The end

Thanks for your attentions