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Ultrastructural Effects of
Matricharia chamomella
Extracts on Cuticle of 3
rd
Larval Instar of
Cx. quinquiefasciatus
73
Figure 4 After 24 hours of treating
Figure 5 After 24 hours of treating
in disruption by separating of basement membrane
from the cell line, or from epidermis cell layer.
By increasing time of expose period (48 hours, Figure
6 and 7) the effect of treatment reach its higher rate
causing damage in cell layer with interruption and the
cuticle space decrease with big change in the structure
of it shown in its different appearance from control
one (the surface of cuticle being edges, sub cuticle
disappeared, separate of En from Ep). These dramatic-
ally changes causing abnormal state of insect life.
Insect cuticle remains the second most widely
distributed material (the first is plant cell wall and
wood). Under experimental conditions the cuticle can
be shown to be extremely sensitive to water content,
suggesting that an important source of stabilization is
H-bonding (Vincent and Ablett, 1988).
Figure 6 After 48 hours of treating
Figure 7 After 48 hours of treating
2 Materials and Methods
Histological study of the cuticle of the third larval
instar of
Cx. quinquefasciatus
.
Lotions and dyes used in the preparation of electron
microscopy samples.
2.1 0.1 M Na-cacodylate buffer (Glauert, 1991)
Solution (a): Dissolve 21.4 grams of in liters of
distilled water.
Solution (b): Add 8.5 mL of concentrated hydrochloric
acid in a liter of distilled water.
Add 500 mL of solution (a) to 41.5 mL of solution (b)
and adjust the pH at 7.2.
2.2 Prepare glutaraldehyde 3% (Hayat, 1970)
Add 12 mL of glutaraldehyde concentration of 25% to
88 mL Organizer 0.1 Molar sodium cacodylate record
according to the previous method.
2.3 Prepare osmium tetroxide 1%
Prepare 1% of the fourth osmium oxide Osmium
tetroxide 1%. Melt 1 gram of crystals fourth osmium
oxide in 100 mL of solution Organizer 0.1 Molar
sodium cacodylate record according to the previous
method.
2.4 Toluidine blue (Bancroft and Steven, 1982)
1. Dissolve 1 gram of borax (Sodium tetra borate) in
100 cm
3
of distilled water.
2. Add 1 gram of Toluidine blue with continuous
stirring.
3. Filter the dye before use.
2.5 Uranyl acetate (Bozzola and Russel, 1992)
1. Dissolve 2 grams of powder uranyl acetate in 100 cm
3
of distilled water.
Journal of Mosquito Research