It is often necessary to examine the male or female genitalia when identifying tortricid adults. The following is a guide for dissecting males and females and preparing slides for examination of genitalia under a compound microscope. Instructions for removing specimens from sticky traps are also provided.
The following materials are needed for most procedures outlined on this page. If you are planning to slide mount the genitalia, you will need standard microscope slides, cover slips, and mounting medium
| Brushes (two, synthetic, No. 0, 00) | Soapy water solution |
| Forceps (two pair, Dumont #5) | Genitalia vials |
| Syracuse watch glasses | Glycerin |
| Petri dishes (glass) | Ethanol (95% or 100%) |
| Potassium hydroxide (KOH, 10%, or pellets) | Stains (optional) |
| Histoclear (citrus oil) | Dry bath heat block (optional) |
| Dissecting microscope | Slide making materials (optional) |
Step 1: After sufficient soaking in KOH, place the abdomen in a Syracuse dish full of water. A drop of soapy water can be used to break the surface tension so that the abdomen does not continually float to the surface. Use two small brushes to manipulate the abdomen.
Step 2: While holding the abdomen down with one brush (illustrated), gently start to remove scales with the other brush. The abdomen can be brushed in any direction, but care should be taken to not tear the abdomen or damage the genitalia. Scales can also be removed from the genitalia while they are still attached to the abdomen.
Step 3: Continue removing scales. At the same time, begin to brush (extend) the genital capsule away from the abdomen. Do not brush too hard to avoid damage to any important morphological structures.
Step 4: Separate the genital capsule from the abdomen. In some taxa, this can be easily accomplished by simply brushing the genitalia out of the abdomen. In other taxa, you will need to carefully separate the genital capsule from the abdomen using forceps. Be careful to not damage any important morphological structures while separating the genitalia and abdomen. Set the genitalia aside for steps 5–6.
Step 5: After the genitalia is removed, continue cleaning scales from the abdomen. Many specimens will also have remains of the digestive system, tracheal system, etc. still inside the abdomen. While holding the abdomen with a brush, gently reach inside of the abdomen with forceps and pull out this material. The goal is to have the abdomen relatively clean of scales and other debris without causing significant damage.
Step 6: In this illustration, the abdomen is as clean as necessary for the staining procedure. Additional scales can be removed after it is stained. Place the abdomen in a dish with dilute stain. Chlorazol black is recommended for most specimens. Do not stain the abdomen too dark; exact staining times will depend on the stain and concentration. If the stain is very dilute, the abdomen can sit in the stain during steps 7–8.
Step 7: Place the genital capsule into a Syracuse dish with clean water. While holding with a brush, gently remove any remaining scales or pieces of the abdomen that may have become detached with the genitalia. Forceps may be required to remove debris. Once the genitalia is clean, it may be stained if desired. Heavy staining of genitalia is not recommended as it obscures morphological features when viewed under a compound scope. In this example the male genitalia is sclerotizedsclerotized:
hardened; usually in reference to larval structures or adult genitalia 
enough that staining is not required.
Step 8: Place the clean genitalia into a small petri dish with 100% ethanol. Using forceps, gently spread the valvaevalva:
an appendage flanking the intromittent organ that is used to clasp the female during copulation
and place the genitalia under a small piece of glass (a broken microscope slide will work). The ethanol dehydrates and stiffens the genitalia so that it retains this position when placed on the microscope slide. Some taxa may be easy to position; others may be very difficult. It is important to work quickly as the genitalia will begin to stiffen in a short period of time.
Step 9: After the abdomen is sufficiently stained, place it into the same Syracuse dish with clean water that was used to clean the genitalia. Brush off the remaining scales being careful not to damage the abdomen. Place the cleaned abdomen into the petri dish with 100% ethanol and flatten it under a small piece of glass.
Step 10: This illustration shows the abdomen and male genitalia positioned under small pieces of glass immersed in 100% ethanol. These structures should be allowed to dehydrate for at least 1 hour, and can be left for several days with no damage as long as the ethanol does not completely evaporate. If possible, it is recommended to allow the abdomen and genitalia to dehydrate overnight, and continue with slide preparation the following day.
Step 11: After the genitalia and abdomen have sufficiently dehydrated in ethanol, they can be slide mounted. Place a drop or two of Euparal onto the center of the slide. Euparal is soluble in alcohol, so the genitalia can be directly moved from the petri dish onto the slide. Using forceps, position the genitalia and abdomen and completely cover them with Euparal. Using a clean pair of forceps, gently drop the cover slip on the Euparal, being careful to not trap air bubbles under the cover slip.
Step 12: Fresh slides should be stored flat and covered to prevent dust and debris from falling onto the fresh Euparal. Euparal can take up to 6 months to completely cure, although slides can be carefully handled after a day. Heating the slide may help to cure the Euparal quicker, although it is not recommended to heat fresh slides as the genitalia and abdomen may shift under the cover slip. Slides should be labeled with the collecting information from the specimen, the determination (if known), and a unique identifier that ties the slide to the specimen. Pinned or otherwise preserved specimens should receive a label with the same unique identifier.
Step 1: After sufficient soaking in KOH, place the abdomen in a Syracuse dish full of water. A drop of soapy water can be used to break the surface tension so that the abdomen does not continually float to the surface. Use two small brushes to manipulate the abdomen.
Step 2: While holding the abdomen down with one brush, gently start removing scales with the other brush. The abdomen can be brushed in any direction, but care should be taken to not tear the abdomen or damage the genitalia. It is helpful to remove scales from the sterigmasterigma:
the sclerotized region surrounding the female ostium bursae 
and posteriorposterior:
after, to the rear, toward anal end 
segments while they are still attached to the abdomen.
Step 3: After the majority of scales are removed from the posteriorposterior:
after, to the rear, toward anal end segments, separate the genitalia from the rest of the abdomen. Using two pairs of forceps, hold the abdomen between the 6th and 7th segments (arrow), and carefully tear between the sclerites. It may be helpful to partially separate one side, and then start on the opposite side.
Step 4: Once there is complete separation between segments 6 and 7, separate the genitalia from the abdomen. Carefully pull the corpus bursaecorpus bursae:
a dilated membranous sac at the anterior end of the bursa copulatrix
out of the abdomen, which is often surrounded by fatty tissue. It is often helpful to pull tissue and debris out of the abdomen at the same time to avoid breaking the ductus bursaeductus bursae:
a membranous tube connecting the ostium bursae to the corpus bursae
.
Step 5: Once the genitalia is removed, continue cleaning scales and debris from the abdomen and genital segments. Many specimens will have remains of the digestive system, tracheal system, fatty tissue, eggs, etc. still inside the abdomen. While holding the abdomen with a brush, gently reach inside of the abdomen with forceps and pull out this material. The goal is to have the abdomen relatively clean of scales and other debris without causing significant damage. After it is relatively clean, the abdomen can be stained. Place the abdomen in a dish with dilute stain. Chlorazol black is recommended for most specimens. Do not stain the abdomen too dark; exact staining times will depend on the stain and concentration. If the stain is very dilute, the abdomen can sit in the stain while the genitalia is cleaned.
Step 6: Continue cleaning the genital segments to remove scales, tissue, and debris. It is often necessary to remove a spermatophore from the corpus bursaecorpus bursae:
a dilated membranous sac at the anterior end of the bursa copulatrix. Using forceps, tear a small hole in the anterioranterior:
before, to the front, toward the head 
end of the corpus bursaecorpus bursae:
a dilated membranous sac at the anterior end of the bursa copulatrix and pull out the spermatophore. Be careful to not completely tear or destroy the corpus burase. Once the genital segments are relatively clean, they can be stained. Light staining with chlorazol black is recommended for most females. The genitalia can be stained in the same dish as the abdomen. Do not stain the abdomen too dark; exact staining times will depend on the stain and concentration.
Step 7: After the genital segments are sufficiently stained, place them into a Syracuse dish with clean water; use a drop of soapy water to break the surface tension. Clean the genitalia of any remaining scales or debris. Clean the inside of the corpus bursaecorpus bursae:
a dilated membranous sac at the anterior end of the bursa copulatrix if any parts of the spermatophore remain.
Step 8: Place the clean genitalia into a small petri dish with 100% ethanol. Flatten the genitalia under a small piece of glass (a broken microscope slide will work). The ethanol dehydrates and stiffens the genitalia so that it retains this position when placed on the microscope slide. Be careful to not smash or heavily distort the papillae analespapillae anales:
the female ovipositor lobes 
. Although female genitalia is much easier to position than male genitalia, it is important to work quickly as the genitalia will begin to stiffen in a short period of time.
Step 9: After the abdomen is sufficently stained, place it into the same Syracuse dish with clean water that was used to clean the genitalia. Brush off the remaining scales being careful not to damage the abdomen. Place the cleaned abdomen into the petri dish with 100% ethanol and flatten it under a small piece of glass.
Step 10: This illustration shows the abdomen and female genitalia positioned under small pieces of glass immersed in 100% ethanol. These structures should be allowed to dehydrate for at least 1 hour, and can be left for several days with no damage as long as the ethanol does not completely evaporate. If possible, it is recommended to allow the abdomen and genitalia to dehydrate overnight, and continue with slide preparation the following day.
Step 11: After the genitalia and abdomen have sufficiently dehydrated in ethanol, they can be slide mounted. Place a drop or two of Euparal onto the center of the slide. Euparal is soluble in alcohol, so the genitalia can be directly moved from the petri dish onto the slide. Using foreceps, position the genitalia and abdomen and completely cover them with Euparal. Using a clean pair of forceps, gently drop the cover slip on the Euparal, being careful to not trap air bubbles under the cover slip.
Step 12: Fresh slides should be stored flat and covered to prevent dust and debris from falling onto the fresh Euparal. Euparal can take up to 6 months to completely cure, although slides can be carefully handled after a day. Heating the slide may help to cure the Euparal quicker, although it is not recommended to heat fresh slides as the genitalia and abdomen may shift under the cover slip. Slides should be labeled with the collecting information from the specimen, the determination (if known), and a unique identifier that ties the slide to the specimen. Pinned or otherwise preserved specimens should receive a label with the same unique identifier.
