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For the cadaveric validation, fifteen fresh human thigh/knee specimens from 10 cadavers (2 females, 8 males) were dissected.
When available, up to 50 males were dissected for the examination of genital structures relevant to identification.
The embryonic kidneys and adult kidneys (from either pregnant or non-pregnant females or males) were dissected in ice-cold Phosphate Buffered Saline (PBS) and fixed overnight in fresh 4% paraformaldehyde in PBS at 4°C.
Around 8 10 males were dissected in ice-cold PBS, treated with collagenase, fixed with 4% paraformaldehyde at room temperature, rinsed and washed with PBS containing 1% Triton-X-100.
A. gambiae adult males were dissected in 1 × PBS (Ambion).
Males were dissected in a small amount of phosphate buffered saline on a microscope slide.
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Bees were chilled on ice and the antennae of each torpid male were dissected by sterile forceps and immediately shock-frozen on liquid nitrogen.
The reproductive tract of each male was dissected into phosphate saline buffer.
If recaptured (37 males), SEVs were dissected out, and the number and robustness of sperm were estimated.
Two-month-old mouse mandibles (four wild-type male and four affected male mice) were dissected following cervical dislocation and fixed in 2% paraformaldehyde/2% glutaraldehyde prepared in 0.1 m cacodylate buffer containing 0.15 m sucrose and 2 m m calcium chloride (pH 7.3) at 4°C overnight.
Male genitalia were dissected by putting them in 10% warm KOH solution for 10 15 min to macerate the abdomen; the rest of the dissected parts was preserved in 70% ethyl alcohol plus few drops of glycerin.
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