=====18.SOP FOR FROZEN SECTION===== 1.** PURPOSE OF EXAMINATION:** To lay down the procedure for Frozen section in Histopathology section at NCHLS Surat. 2.** PRINCIPLE AND METHOD OF PROCEDURE USED FOR EXAMINATION:** When tissue is rapidly cooled down, water contained in it converts into ice which act as fixatives, supportive media and enable the cutting of thin section. 3.** STAFF RESPONSIBLE:** lab. Technician under supervision of resident doctor/tutor. 4.** PERFORMANCE CHARACTERISTICS:** NA 5.** TYPE OF SAMPLE:** Frozen section 6.** PATIENTS PREPARATIONS:** NA 7.** TYPE OF CONTAINER AND ADDITITVES:** Clean labelled containers, jars 8.** REQUIRED EQUIPMENTS AND REAGENTS:** **EQUIPMENTS:** Cryostat **REAGENT:** Normal saline, H&E stain, absolute alcohol, Xylene, DPX. 9.** ENVIRONMENTAL AND SAFETY CONTROL:** Universal safety precautions 10.** CALIBRATION PROCEDURE (METROLOGICAL TACEBILITY):** temperature measurement and calibration for section thickness done by manufacturer. 11.** PROCEDURE (STEPS):** • When frozen section is demanded by clinician respective authorized signatories are informed for frozen section and approximate time. • Cryostat is switched on the previous night(5-6 hrs before) and temperature is noted. • When specimen comes for frozen section, patient’s identity is checked. • The specimen is then given a frozen no.(eg. F-10) which is also tagged to the patient’s requisition form. • If more than one specimen of the same patient is sent for frozen section, they are labelled in alphabetical order (eg. FS1A, FS1B, FS1C...) • Grossing of specimen done and representative sections are submitted for freezing in cryostat. • Place the representative tissue on a metal cryostat chuck after applying 1-2drops of tissue adherent (gung) on the chuck. • Pour 1-2 drops of normal saline on the tissue. • Put the chuck with the tissue on a metal tray of the cryostat for freezing the tissue. • When the temperature of the cryostat attains -250C to -300C, trim the tissue to get full exposure. • Cut the section at 5u thickness and pick up from the knife to put it on previously labelled glass slides. • Sections are fixed in oven for 2minutes at 500C to 600C. • Sections are then stained with H&E stain. • Reporting of frozen section is done by authorized signatories for frozen section(professor/additional professor) • The report is informed telephonically to concerned surgeon after verifying the identity of the patient and the person to whom it is informed. • Same written report is dispatched immediately. • Entry of diagnosis,date,and name of reporting pathologists, technicians, TAT and telephonic conveyance of report to surgeon is written down in the frozen register. • Remaining tissue bit and uncut thawed frozen bits are placed in a container with 10% buffered formalin. A HP no. Is then given to this specimen. • When final diagnosis is made, it is entered against its frozen diagnosis in frozen register. • The frozen section slides are stored separately in storage cabinets. • After completion of procedure, cryostat is cleaned with Hypochlorite solution, trimming and blade are removed and oiling of machine is done. 12.** QUALITY CONTROL PROCEDURE:** Section is examined for proper staining, processing and same sections are reviewed by another authorized signatories. 13.** INTERFERENCE:** temperature of cryostat 14.** PRINCIPLE OF PROCEDURE FOR CALCULATING RESULTS INCLUDING, WHERE RELEVENT, THE MEASUREMENT OF UNCERTAINITY OF MEASURED QUANTITY VALUES:**NA 15.** BIOLOGICAL REFERENCE INTERVALS OR CLINICAL DECISION VALUES:**NA 16.** REPORTABLE INTERVAL OF EXAMINATION RESULTS:** within 30 minute of receipt of tissue. 17.** INSTRUCTION FOR DETERMINING QUANTITATIVE RESULTS WHEN RESULTS IS NOT WITHIN THE MEASUREMENT INTERVAL:** NA 18.** ALERT /CRITICAL VALUES, WHERE APPROPRIATE:** NA 19.** LABORATORY CLINICAL INTERPRETATION:** NA. 20.** POTENTIAL SOURCE OF VARIATION: **NA 21.** REFERENCE:** Histopathological laboratory techniques by Culling’s.