To calculate number of egg masses, roots were washed with clean water and stained in 0.1% acid
fuchsine solution (acid
fuchsine dissolved in a 1:1:1 mixture of glycerol, lactic acid and distilled water) for 1-3 min.
Semithin sections were stained with Epoxy Tissue Stain (EMS, USA), containing toluidine blue and basic
fuchsine. Ultrathin sections were mounted on formvar coated grids and double stained with 0.5% uranyl acetate and 3% lead citrate.
It might consist of a combination of different triphenyl-methane dyes such as
fuchsine and crystal violet, solvents and mordants like ethanol, acetone and lugol's iodine as reagents of Gram staining as the most extensively performed test in microbiology.
The slide-mounting process was as follows: (a) after making a small incision on the back of papaya mealybug, it was heated in 10% NaOH for 10 min; (b) the body contents was expelled and the body was gently flattened with a microspatula; (c) the preparation was stained morphologically in a saturated solution of
fuchsine in a mixture of distilled water, lactic acid and glycerol (1:1:1) for 3-5 h; (d) washed in glacial acetic acid for 1 h to stabilize the staining, transferred to lavender oil, allowed to stand for at least 1 h, and (f) placed in a drop of Canada balsam on a slide and covered with a cover glass.
Transversal freehand sections were made at middle third of the blade, followed by staining with basic
fuchsine diluted to 0.5% in ethanol 50% and Astra blue, to show the lignified walls and the cellulose walls in the leaf cells.
Teeth from both groups were then immersed in 2% basic
fuchsine dye for 8 hours separately.
Slides were stained with toluidine blue and acid
fuchsine and were analyzed using a microscope connected to a video camera interfaced to a computer, where specific processing software was used for measurements (ImageJ 1.34, National Institutes of Health, Bethesda, MA, USA).