Histopathologic Techniques

The art and science performed by the histotechnologist that will aid the pathologist in order to diagnose or rule out disease

Biopsy Excision entire organ Incision part of an organ

Autopsy

External Growth Surgical growth growth of an organ Endoscopic growth unusual growth

Stages of death 1. 2. 3. 4. 5. Algor mortis change in temperature cold Rigor mortis hardening of the tissue Livor mortis Change in color Autolysis Putrefaction

Methods of Fresh Tissue Examination

1. Teasing or Dissociation dissected or dissociated within a watch glass containing isotonic salt solution

2. Squash preparation or Crushing small pieces of tissues not more than 1 mm in diameter are forcibly compressed using slide or coverslip 3. Smear Preparation a. Streaking using applicator stick or loop. Direct or zigzag line with a relatively uniform distribution of secretion b. Spreading selected portion is placed on a slide and gently spread throughout the slide c. Pull apart using a slide, a drop of sample on the end, another slide will be placed on the specimen, using a gentle motion, let the specimen to spread to both slides, and with uninterrupted motion pull apart the slides. d. Touch Preparation or Impression Smear the tissue will be lightly pressed on the slide and be removed 4. Frozen Section rapid diagnosis especially for lipids and nervous tissue

Processing of Tissue 1. Fixation 2. Dehydration 3. Clearing 4. Infiltration 5. Embedding (Impregnation) 6. Trimming 7. Section-cutting 8. Staining 9. Mounting 10. Labeling

Fixation The first and crucial step in tissue processing Primary goal to maintain the integrity of the specimen as if it is in a life-like manner Secondary goal to protect the tissue from the damage of further handling

Two Mechanisms of action of Fixation

1. Additive it is where the fixative solution itself forms cross-linkages to prevent autolysis furthermore making it unsuitable for putrefaction 2. Non-Additive it is where the fixative solution is not incorporated on the tissue, rather, it alters the chemical composition of the tissue such as removal of water and forming of cross-linkages that would stabilize the specimen