FROZEN SECTION PROCEDURE
(INTRAOPERATIVE CONSULTATION).
DRAFT COPY ONLY.
(Procedure 25).

Gladys L. G. Alonsozana, MD.
Chief, Surgical Pathology Section.
Chief, Gynecologic Pathology Section.
http://www.netautopsy.org/axsop/axsop025.htm


NEXT PAGE
PREVIOUS PAGE
RETURN TO TABLE OF CONTENTS

United States Government Work, uncopyrighted, public-domain, DRAFT COPY ONLY. This document does not necessarily represent the views or policies of any United States Government agency. This document is provided "as is", without warranty of any kind, express or implied, including but not limited to the warranties of merchantability, fitness for a particular purpose and non-infringement. In no event shall the authors be liable for any claim, damages or other liability, whether in an action of contract, tort or otherwise, arising from, out of, or in connection with the document or the use or other dealings made with the document.

See also: Frozen Section Policies: 24, 26, 27, 41, 86.



PRINCIPLE OF THE TEST.

Frozen sections are intraoperative consultations used to establish a rapid histopathologic diagnosis of a pathologic process. Other indications for this special consultation include assessment of surgical resection margins, apportioning tissue for special studies, and harvesting fresh or snap-frozen tissue for research studies.

Fresh, unfixed tissue is frozen to make it hard enough to cut thin sections. A cryostat, a special microtome refrigerated to -20oC is then used to cut a frozen section, which is placed onto a glass slide, stained, and cover-slipped for examination under the microscope.



SPECIMEN REQUIRED.

All fresh human tissue excised at surgery, along with a filled-out Tissue Examination Form (U. S. Standard Form 515, USSF515), with a request for frozen section on the form, and personally handed to a histology technician, a resident, or an attending pathologist. The specimen may be frozen or occasionally examined grossly only, and submitted for permanent sections (see Procedure 24, step #9).



REAGENTS, INSTRUMENTATION.

Cryostat. O.C.T. compound. Hematoxylin and Eosin stain. Glass slides. MM24 (mounting media).



STEP-BY-STEP DESCRIPTION.

1. When the specimen arrives, it is immediately assigned an accession number, which is entered on the Tissue Examination Form, and into the HISTOLOGY LOGBOOK in the histology laboratory, 4D-124. The accession number is also entered into the FROZEN SECTION LOGBOOK in the anatomic pathology signout room, and the time of receipt is recorded.

2. The frozen section pathologist must be informed of receipt of the frozen section specimen (i.e., if the frozen is received by either the resident or histology technician), before the specimen can be processed.

3. Under the guidance of the pathologist, the specimen is measured, described, and dissected (if necessary).

4. Select the appropriate size Cryomold, and fill the Cryomold cavity halfway with O.C.T. Compound.

5. Orient the tissue so that it is positioned in the center of the mold. Add O.C.T. Compound flush with, or just slightly over, the second level of the Cryomold cavity.

6. Place the Specimen Holder onto the Cryomold, and press down lightly.

7. Place the Specimen Holder/Cryomold assembly, with the Cryomold down, into one of the wells on the Cryobar. If desired, place the Heat Extractor on the Specimen Holder, to accelerate the cooling process.

8. Allow the O.C.T. Compound to freeze completely. ( Note: Complete freezing of the O.C.T. Compound can be checked by removing and inspecting the Specimen Holder/Cryomold assembly, to see if the O.C.T. Compound is completely white in color. The time it takes to freeze completely depends primarily upon the size of the Cryomold.)

9. When the frozen specimen block is ready for use, lift the Heat Extractor (if used) from the Specimen Holder, and slide it out of the way. Remove the Specimen Holder/ Cryomold assembly from the cooling unit.

10. Grasp the tab of the Cryomold with the thumb and forefinger, and pull it away from the specimen block. Set aside or discard the Cryomold.

11. Place the specimen block (frozen specimen on the Specimen Holder) into the Microtome Chuck, and turn the Specimen Holder Knob clockwise (toward you) to secure the specimen block.

12. If necessary, position the Blade Holder and/or the Chuck to the desired angle.

13. Lift the Blade Guard away from the blade.

14. Press the Motorized Retraction key (up arrow, ↑) to retract the specimen block (move the block away from the blade); or Press the Motorized Advance key (down arrow, ↓) to move the specimen block toward the blade. (Note: Pressing and holding the (Motorized retraction (↑)/ Motorized advance (↓)) key will continuously retract/forward the specimen block away/toward the blade, respectively.)

15. Perform AUTOMATED TRIMMING by pressing the intermittent motorized sectioning or continuous motorized sectioning key. Once motorized sectioning is activated, press and hold the (Trim) key on the left control panel to begin sectioning. Each section is one trim thickness. Release the (Trim) key to resume regular sectioning.

16. Cut and discard several sections.

17. Use a small brush to apply light pressure to the leading edge of the section, then gently pull the section forward onto the Clamping Plate. (Note: Alternatively, use the Anti-Roll Rake or Anti-Roll Plate to help obtain flat sections).

18.Examine a section to determine if the section quality is acceptable.

19. Mount the specimen section by touching it with a clean and properly labeled microscope slide.

20. Stain the slide manually, following the H&E Frozen Section Staining Procedure, posted above the staining solutions. Coverslip.

21. Deliver the completed slides to the signout room, 4D-139, for the attending pathologist to read.

22. When a diagnosis has been rendered, the attending pathologist records the diagnosis on the Tissue Examination Form, initials the diagnosis, and the Operating Room is immediately called with the report, either by the resident pathologist or by the attending pathologist. It is expected that completion time will occur within 20 minutes per slide from the time of receipt of the specimen.

23. The patient's identification is checked and confirmed before delivery of any verbal report. At the start of the call, the pathologist repeats the name of the patient to the surgeon.

24. The pathologist should asked the surgeon to read back the report, and a READ BACK statement must be entered in the requisition form along with the date and time the diagnosis was rendered. Read-back statistics are reported monthly to the Invasive Procedures Review Committee. See: Procedure 138. Invasive Procedures Review Committee. and Procedure 238. Read Back.

25. Record the completion time in the FROZEN SECTION LOGBOOK in the pathology sign out room, 4D-139.

26. Once the diagnosis is reported, and with approval of the attending pathologist, the frozen section specimen is processed for permanent section. The original handwritten report is kept in the office along with the written submission sheet. In addition, the report is added to the permanent GROSS DESCRIPTION section of the final (permanent) report that appears in the patient's chart.

27. FROZEN SECTION TURNAROUND TIME PILOT STUDY. A Frozen Section Turnaround time pilot study was performed for the period March 1, 2001, through December 31, 2001. During this time, there were 360 frozen section reports recorded in the frozen section. In the same period, there were 4,134 cases from year 2000 and 4,920 cases from year 2001. Thus, frozen section reports were issued for 4% of cases. For a single case (0.3%), the report was available only after 22 minutes of receiving tissue from the operating room.



REFERENCE.

1. Prophet EB, Mills B, Arrington JB, Sobin LH.
Laboratory Methods in Histotechnology, pp. 67-70.
1992: Armed Forces Institute of Pathology, Washington, DC. 20306-6000.
ISBN: 1-881041-00-X 1992.