LAVALLE, Jestin B.

BMLS Intern Histopathology

Topic 2: Fresh Tissue Preparation

1. Purpose of fresh tissue preparation.

To examine the tissue in the living state, allowing protoplasmic activities such as motion, mitosis, phagocytosis and pinocytosis to be
observed

Limited use because of changes after death (temporary)

2. 7 methods of fresh tissue preparation? Explain each.

Teasing / Dissociation – Immersed in a watch glass w/ NSS, carefully dissect or separate, examine:
o Phase contrast
o Bright field – may stained w/ methyl violet or methylene blue
Squash Preparation (Crushing) – small piece (1 mm) is placed in a slide and forcibly compressed with another slide or cover
glass
o Vital stain may be placed at the junction of the slide and cover glass, absorbed through capillary attraction
Streaking – Use of applicator stick or a platinum loop, rapidly and gently applied in a direct or zigzag line in a relatively uniform
distribution
Spreading – Selected portion is transferred and gently spread into a moderately thick film (circular motion)
Pull-Apart – A drop of a secretion or sediment is placed upon the slide Another clean slide is placed over allowing to disperse
evenly over the surface of the two slides. Slide are the pulled apart with a single uninterrupted motion
Touch Preparation (Impression smear) – Surface of a freshly cut piece of tissue is brought into contact and pressed on to surface
of a clean glass slide
o Phase or Light
o Cells are examined without destroying their intercellular relationship, and without separating them from their normal
surroundings
Frozen Section – Slices (10-15um) are cut from a fresh tissue frozen on a microtome with CO2 or a cold chamber
o Recommended in lipid and nervous tissue demonstration

3. Enumerate 9 cellular degenerative changes.

4. Explain each.

o Hypoplasia – incomplete growth of an organ

o Atrophy – when the mass of the functioning cells in a tissue is reduced
o Agenesia – complete failure of growth of an organ during embryological development
o Metaplasia – cells adapt to change in environment by altering their morphological appearance
o Dysplasia – increased rate of cell division is coupled with incomplete maturation of resultant cell (High N:C)
o Neoplasm – abnormal proliferation of autonomous cells serving no purpose of growing
o Aplasia – failure of differentiation of embryonic tissue into a developed organ
o Anaplasia – regressive change in adults cells toward more primitive
o Atresia – failure to form an opening

5. What are the 5 Cardinal Signs of Inflammation?

 Redness (Rubor) – Blood vessel dilate
 Swelling (Tumor) – Plasma exists vessel fluid exudation
 Heat (Calor) – BV dilate
 Pain (Dolor) – release of bradykinin
LAVALLE, Jestin B. BMLS Intern Histopathology
 Loss of function (Functio laesa) – combined effect of all

6. What is somatic death?

Irreversible unconsciousness, Death of entire body

7. Enumerate the primary changes during somatic death.

 Cessation of cardiac function – flat ECG
 Cessation of pulmonary function
 Cessation of nervous function – flat EEG

8. Enumerate the secondary changes after somatic death.

a. Algor mortis – Cooling of the body; Absence of circulation

b. Rigor mortis – Stiffening of the body; Soluble to insoluble muscle protein (actin, myosin)
c. Livor mortis – Purplish discoloration
d. Post portem clotting – Clotting (blood flow stops)
e. Drying – Dryness and wrinkling
f. Putrefaction – Softening of body, lysing of cells (foul odor, decay, purplish); bacteria accumulate
g. Autolysis – Lysis of cells; Lysosome (suicide sac)

9. What is autopsy? Purpose?

Examination of a dead body to find out the cause of death; “Autopsia” – to see with one own’s eye

o To interpret and correlate facts surrounding death

o Establish time of death
o Evaluate any injury or disease present

10. What are the 2 types of autopsy? Differentiate.

Forensic – determine the cause and manner of death, obtains biological specimen
Ex. Police investigation, Toxicology if poisoned

Clinical – more insights in pathological process and can be prevented in future

Ex. Research and educational

11. What are the four autopsy techniques? Differentiate.

o Virchow technique
o Rokitansky technique
o Le tulle (En masse) technique
o En bloc (Ghon/Zenker) technique