Surgeon's Collection Protocol - TESE & microTESE

Collection PROTOCOL: TESE and microTESE

This protocol outlines the collection process for Testicular Sperm Extraction (TESE) and Microsurgical Dissection Testicular Sperm Extraction (microTESE) performed by a surgeon trained and experienced in male reproductive surgery. These procedures involve the surgical retrieval of sperm from the testis and are typically conducted in a hospital operating room (OR) or an ambulatory surgery center. Prior to the procedure, it is crucial to verify that the required materials and equipment are prepared, including the sperm-washing media, sterile containers, and patient identification labels. This guide details each step, from specimen collection to transport, ensuring proper handling and processing for optimal results.

KIT CONTENTS – Please verify the contents before the procedure

1. Sperm Washing Media – One sealed 12 ml container: Place in refrigerator upon receiving. Leave at room temperature at least one half hour prior to procedure.
2. Two-15 ml Conical Tubes used for TESE specimens from Right and Left Testis
3. Eight Patient Labels – Four labeled LEFT Testis and Four Labeled RIGHT Testis
   1. Have the Patient confirm the information and initial each label
   2. Place a label on each container/conical tube into which a specimen will be placed. Note that one container/conical tube should have a label with LEFT and another with RIGHT.
4. Two sterile glass Petri dishes for mincing the testis tissue. After use, have the OR clean, sterilize, and give back to you for your next case.
5. Slide Holders
   1. Right and Left Gross Biopsy - 2 slides – 10 coverslips
   2. Right microTESE – 4 slides (pre-labeled) - microTESE only
   3. Left microTESE - 4 slides (pre-labeled) - microTESE only
6. Eight 1mL cryovials are included in the small white box- microTESE only
7. Two sterile urine specimen containers as an alternative for mincing the testis tissue. Discard if not used.
8. A specimen bag to hold the conical tubes with specimen and sperm washing media upright for transport. Once media and/or specimens are placed in the conical tube, keep the tubes upright to prevent leakage of the specimen. You should wrap the conical tubes in clean OR towels before placing back in the Transport box prior to transport back to New York Cryo for processing.
9. A second specimen bag to place the rinsed glass Petri dishes after use

DISPOSABLES

• Sterile glass petri dishes - supplied in the Kit
• 1ml Disposable syringes – supplied by the OR - one for each specimen returned to the lab (two for a TESE and 10 for a microTESE)
• Two 15mL Falcon conical tubes - supplied in the Kit
• Patient Identification labels- supplied in the Kit
• Sterile glass slides with cover slips - supplied in the Kit

EQUIPMENT:

1. Operating microscope
2. Phase contrast microscope
3. 37⁰ Heating block for Conical tube and cryovials (if available)

Testicular Sperm Extraction (TESE) and Microsurgical Dissection Testicular Sperm Extraction (microTESE)

 General overview of the procedure

• Surgical retrieval of sperm by microTESE
• The procedures may take place in the hospital OR or an Ambulatory Surgery center
• On the day of surgery, a 12 mL vial of Sperm Wash Medium, sterile 15mL Falcon conical tubes.
• A consent for TESE sperm freezing will be obtained from the patient by the laboratory usually after specimens are received by the laboratory.
• Once in the OR, obtain a hospital-printed identification stickers from the OR staff and confirm the identity of the patient with both the physician and the OR staff.  This Hospital label will be affixed to the microTese Intraoperative Worksheet.

Intraoperative sperm identification

• Multiple small, sterile, Petri dish (‘DISH’) with 500 ul of sperm washing media (SWM) in each dish for microTESE and 1000 ul (1 ml) in each dish for TESE. Sterile glass petri dishes are preferred, as plastic petri dishes can have plastic shavings removed from the bottom surface during the dissection process.
• NOTE: The surgeon’s preference might be to group several regions of the testis together and place and place the specimens obtained into a single 15 cc Conical tube. In this case, 1 ml of SWM should be placed in the petri dish and 1 ml in the Conical tube (2ml total volume of SWM). The surgeon will identify where the specimens in each conical tube have been taken from (Figure 1 & Figure 2) on the log sheet and conical tube.
• The Surgeon will place 1 or more TESE/microTESE specimens in a single SPECIMEN DISH and specify the location the specimen was taken from to be indicated on the microTESE Worksheet (”LOG”) in accordance with the locations as specified in Figure 1 & Figure 2.  The specimen dish should be labeled with the locations also.
•  The specimen is processed by mincing into small pieces. Ideally, the processed specimen should be able to pass through a 24 gauge Angiocath (yellow); A small portion of the specimen will be examined on a clean slide with a small square coverslip (22x22 mm) under 400x with a Phase contrast microscope in the OR. The slide will be labeled with the number that corresponds to the sample on the LOG sheet with the abbreviation for the location of the testis from which it was taken.  The andrologist/surgeon must make sure that when the slide is made that a small amount of the tissue from the sample is transferred onto the slide so it can be viewed.  After a satisfactory slide has been made, the remaining tissue sample should be immediately to the appropriately labeled conical tube using the 1 ml syringe. The LOG sheet should be updated with the microscopic quality of the specimen as viewed in the operating room
• The specimens will be identified on the LOG sheet by Specimen number, testis Location, Quantification of Sperm and sperm motility (Figure 1 & Figure 2).
• The Andrologist/Surgeon will determine when to end a microTESE procedure based on intraoperative findings.

Transport to the Andrology Laboratory

• All specimens should be transported to the laboratory as soon as practical. The conical tube should be closed securely, kept upright to prevent leakage of the tissue and media, and placed in a specimen bag. The specimens should be kept as close to room temperature as possible during transport.They can be wrapped in a towel and placed in a temperature protected box for transport to the Laboratory.

FIGURE 1

Figure 2

Please call the office (516-487-2700) or Dr. Bruce Gilbert's cell (516-595-9475) with any questions.