TITLE: Post-Surgical Dissection of Cervix
AUTHORS: Stephen Fisher, Marielena Grijalva, Rong Guo, sarahjoh, Hieu Nguyen, John Renz, Jean G Rosario, Steven Rudich, Brian Gregory, Junhyong Kim, Kate O'Neill

[DESCRIPTION]
This protocol describes dissection of the ectocervix and endocervix in preparation for 10X Visium, 10X Multiomics, pathology review, and biobanking.  The cervix is the cylindrical lower part of the uterus which connects the uterus to the vagina.  It is typically 3-4cm in length is largely fibromuscular in structure.  The endocervical canal is lined with a thin layer of glandular epithelium.  In this protocol we debulk the fibromuscular portion of the cervix for endocervical samples in order to ensure cells lining the endocervical canal are represented.

[BEFORE_START]
Dilute phosphate buffered saline (Gibco; 14200-075) to 1X with nuclease-free water.

[STEPS]
SECTION: Dissection of Ectocervix
1. Remove uterus from ice and dry the cervix.
SECTION: Dissection of Endocervix
12. Remove uterus and remaining cervix from ice and dry.

SECTION: Dissection of Ectocervix
2. Using a large grossing knife with a fresh blade, remove the most distal portion (1cm length) of the cervix which contains the ectocervix
SECTION: Dissection of Ectocervix
3. Place the remainder of the cervix and uterus back on ice.
SECTION: Dissection of Ectocervix
4. Weigh and measure this distal portion of cervix.
SECTION: Dissection of Ectocervix
5. Lay down cervix with the external cervical os facing up, with the 12 o’clock anterior position furthest away from you.

  

SECTION: Dissection of Ectocervix
6. Dry the ectocervix and label with marking dyes at 12 o’clock, 3 o’clock, 6 o’clock, and 9 o’clock.
SECTION: Dissection of Ectocervix
7. Moving clockwise, slice the cervix vertically from 12 to 6 o’clock and horizontally from 9 to 3 o’clock so there are 4 equally sized pieces.
SECTION: Dissection of Ectocervix
8. Divide each piece into four additional sections radially.
SECTION: Dissection of Ectocervix
9. Using disposable weigh boats, weigh each tissue specimen.
SECTION: Dissection of Ectocervix
10. For later 3D anatomical positioning, track the quadrant and radial section of each piece of tissue.
SECTION: Dissection of Ectocervix
11. Tissue can be processed with protocols OCT-Embedded Tissue Preparation, Tissue Fixation Preparation, or Snap-Frozen Tissue Preparation depending on the desired downstream processing.
SECTION: Dissection of Endocervix
13. Using a large grossing knife with a fresh blade, remove the most distal 1cm of the cervix (previously the middle 1cm of cervix, prior to removal of ectocervical sample).
SECTION: Dissection of Endocervix
14. Return uterus to ice.
SECTION: Dissection of Endocervix
15. Trim ~1/2 of the eccentric cervical stroma.
SECTION: Dissection of Endocervix
16. Dry the endocervix and mark using marking dyes at 12 o’clock, 3 o’clock, 6 o’clock, and 9 o’clock.
SECTION: Dissection of Endocervix
17. Lay the cervix down with the external cervical os facing up, with the 12 o’clock anterior position furthest away from you.
SECTION: Dissection of Endocervix
18. Moving clockwise, slice the cervix in sections as was done above with ectocervix.
SECTION: Dissection of Endocervix
19. Divide each section into 4 additional sections radially.

  

SECTION: Dissection of Endocervix
20. Using disposable weigh boats, take the weight of each tissue specimen.
SECTION: Dissection of Endocervix
21. For later 3D anatomical positioning, track the quadrant and radial section of each piece of tissue.
SECTION: Dissection of Endocervix
22. Tissue can be processed with protocols OCT-Embedded Tissue Preparation, Tissue Fixation Preparation, or Snap-Frozen Tissue Preparation depending on the desired downstream processing.