STANDARD OPERATING PROCEDURE FOR SECTIONING, STAINING AND PROCESSING TISSUE SAMPLES FOR HISTOPATHOLOGY EVALUATION

1.0 PURPOSE

To cut thin sections from tissue block. Good quality cutting of section and staining is a prerequisite for satisfactory morphological examination of tissue without good quality section chances of missing out on findings and erroneous reporting by the pathologist increase.

2.0 SCOPE

This SOP shall be applicable to all biopsy specimens received. This contains the procedures for all the technical staff in the section.

3.0 RESPONSIBILITY

3.1 Histopathology Department Personnel, Head and Quality Assurance Person. To execute review processes for SOP. To maintain the original copy of the SOP. To issue a control copy of the SOP. To maintain the issuance record of SOP and to retrieve the control copy of the SOP & maintain its record.

3.4 Quality Assurance Head: To approve new or revised SOP. To ensure the implementation of the defined system. To assign work to QA executive & department.

4.0 DISTRIBUTION

4.1 The Quality Assurance department is responsible for keeping SOP ‘Master Copy’ approved through the Quality Assurance Head.

5.2 The copy of ‘Control Copy’ of SOP is being distributed in Histopathology departments and placed near related Equipment/Instruments.

6.0 ABBREVIATION(S)

6.1 Abbreviation(s)

6.1.1 SOP: Standard Operating Procedure

6.1.2 No. : Number

6.1.3 Dept. : Department

6.1.4 QA: Quality Assurance Person or Head

6.1.5 Next Rev: Next Revision

6.1.6 Rev. : Revision/Review

7.0 PROCEDURE

· INSTRUCTION	· RESPONSIBILITY
· The sections are now ready for further staining.	· Technical staff
· Daily temperature log of tissue floatation water bath is maintained.	· Technical staff
· At the beginning of the daily run, one tissue section is reviewed for quality of processing, embedding, cutting, and staining. The rest of the section is stained after results are found acceptable or after performing required corrective action.	· Technical staff& · Technical staff · Pathologist.
· Paraffin block & slide are retained for 9 years at room temperature.	· Technical staff

7.1 Requirements

7.1.1 Slide warming table

7.1.2 Tissue float bath

7.1.3 Disposable blades

7.1.4 1% Hydrochloric acid

7.1.5 10% Formic acid

7.1.6 Egg albumin

7.1.7 Distilled water

7.2 Trimming of blocks

7.2.1 Trimming knife is fixed to the knife holder, in such a position that there is a 3-6 degree clearance angle.

7.2.2 The cutting surface of the block is placed parallel to the edge of the knife.

7.2.3 The thickness indicator of the microtome is adjusted to 10-30 microns.

7.2.4 Trimming of the block is done till the entire surface of the tissue to be cut is exposed. The block is removed and placed on ice, with the cutting surface downwards for cooling.

7.3 Section Cutting

7.3.1 The microtome thickness adjusts to 3-4 microns.

7.3.2 The block is removed from the ice and fixed to the block holder of the microtome.

7.3.3 Ribbons of sections are cut usually by about 3-4 microns for

7.3.4 The free end of the ribbon is held with a fine brush and then transferred into the pre-warmed floatation bath.

7.3.5 The tail end of the ribbon is placed in contact with the water in the bath. The section should be floated with the smooth surface facing down.

7.3.6 30 sec-1 minutes is allowed for the section to flatten out.

7.3.7 The flattened sections are drawn onto adhesive-coated pre-numbered slides.

7.3.8 The slides with the sections are placed on the pre-warmed slide warmer for drying for 10-15 minutes.

8.0 PRECAUTIONS

8.1 Handel knives with care to prevent injury.

8.2 Blot the surface of the water n the floatation bath after each block to prevent floaters.

9.0 REFERENCE(S)

9.1 User Manual Microtome

END OF THE DOCUMENT

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