How to access a tick's uterus?

How to access a tick's uterus? - briefly

Place the tick on a chilled slide, make a precise dorsal incision along the abdomen with a fine scalpel, and locate the uterus adjacent to the midgut. Gently lift the uterus out using micro‑forceps.

How to access a tick's uterus? - in detail

Accessing the uterus of a tick requires careful preparation, appropriate equipment, and strict adherence to sterile technique. The procedure is typically performed under a stereomicroscope to visualize the minute structures.

1. Specimen selection

   • Choose engorged females, as the uterine sac is enlarged and more visible.
   • Verify that the tick is alive or freshly killed to prevent tissue degradation.

2. Equipment

   • Stereomicroscope with 20–40× magnification.
   • Fine forceps (Dumont #5) and micro‑scissors.
   • Dissection pins, silicone-coated Petri dish, and a drop of phosphate‑buffered saline (PBS).
   • Optional: micro‑probe for gentle tissue separation.

3. Preparation

   • Immobilize the tick ventral side up on a silicone‑coated surface using two pins through the legs.
   • Apply a small PBS droplet to keep tissues moist and to improve optical clarity.

4. Incision

   • Using micro‑scissors, make a longitudinal cut along the dorsal midline, starting just posterior to the scutum and extending to the posterior margin.
   • Avoid cutting too deep to prevent damage to internal organs.

5. Exposure of the reproductive tract

   • Gently separate the cut edges with forceps, exposing the fat body and underlying organs.
   • Identify the paired ovaries; they appear as elongated, pale structures adjacent to the midgut.

6. Isolation of the uterine sac

   • Locate the thin, translucent tube connecting the ovaries to the genital opening.
   • Using a micro‑probe, gently tease the tube away from surrounding tissue, taking care not to rupture it.

7. Extraction

   • Once the tube is freed, lift it with forceps and pull it toward the genital aperture.
   • The uterus can then be removed in one piece or opened to retrieve its contents, depending on experimental goals.

8. Post‑extraction handling

   • Transfer the uterus to a pre‑labeled micro‑tube containing appropriate fixative (e.g., 4 % paraformaldehyde) for histological analysis or to a fresh PBS drop for molecular work.
   • Clean all instruments with ethanol and sterilize before reuse.

Precautions

• Perform the dissection in a laminar flow hood to minimize contamination.
• Maintain the specimen at 4 °C if delays between steps exceed five minutes.
• Dispose of biological waste according to institutional biosafety guidelines.

Following these steps yields a cleanly extracted uterine sac suitable for morphological examination, gene expression studies, or pathogen detection.