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TECHNICAL EVALUATION OF SECTIONING

1A. TECHNICAL EVALUATION OF SECTIONING

Materials:
Per student 1 paraffin block of mystery tissue X.

Procedure:
1. Using the detailed instructions on the use of the rotary microtome (Appendix 1). carefully clamp your paraffin block in the chuck of the microtome and proceed to expose the full face of the tissue in the block by rough trimming. Set the microtome to 10-15 ɥm whilst trimming. Change to the other end of the disposable blade and proceed until you are ready to cut a ribbon of high quality 4 ɥm thick sections

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2. Carefully cut a ribbon of 10 sections and lay it on the black cutting board. If you cannot manage a ribbon of 10 sections then a shorter ribbon will do.

3. Ensure that the waterbath is between 42-48°C before floating sections.

4. With a scalpel carefully divide the ribbon to produce several groups of 3 sections. Transfer one of these groups on to the water bath. Ensure the sections have expanded fully (do not leave them too long or they will over expand).

5. Float your sections for no longer than 20 seconds Within this timeframe select the 2 best sections and using forceps. separate the sections. mount them on two separate slides.

6. Label your sections with your name, group and tissue type and place them in the slide rack provided where they will subsequently be dried at 60°C for 30min. Make sure you retain your block for assessment with the stained, labelled slide.

7. The laboratory staff will stain these sections, so that you can only assess your cutting skills

8. The following week you will receive your H&E stained slides back from the laboratory technical staff. Label your stained slide appropriately with a permanent label. You are to complete a written report for this exercise focusing on section quality.

Question:
List and describe 5 problems that you may have encountered during this practical which could have lead to poor section quality. Give detailed responses how you may have overcome these issues.

Stained slides must be submitted with written reports for assessment.

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