HISTOLOGY & EMBRYOLOGY

Teaching PPT

Dept. of Anat., Hist. & Embry. School of Medicine Xian Jiaotong University

HISTOLOGY
Qiu Shudong

INTRODUCTION
Definition: A science: study normal microstructure & its related function of human body. 4 structural levels: Cell: the smallest structural & functional unit. Tissue: groups of cells (similar in morphology or related in function)intercellular materials 4 types of fundamental tissue epithelium connective tissue muscular tissue nervous tissue

Organs: organizations of various kinds of tissues in particular ways & perform a specific function.
System: formed by several functionrelated organs which together perform a continuous physiological function. For example: digestive system

Why to study histology ?

To complete the knowledge of human bodys structures----from gross to microscopic Be able to understand how the different tissues function----the basis of physiology Can find the diseases only after the normal is known----the basis of pathology It is related to some modern science fields: cell apoptosis, cell recognition, implantation of embryo stem cells, eugenics and etc. It is also a foundation of clinic sciencesfor a good doctor needed in futrue

Unit used in microscope 1m=1/1000 mm 1 n m=1/1000m Maximum resolution Light microscope: 0.2m Transmission electron microscope: 0.2nm Scanning electron microscope: 5nm

Investigative methods of histology

I. Light microscopy

1. Tissue preparation
A. paraffin section preparation Specimen: as fresh as possible Fixation: fixative: formalin solution; purpose: to preserve the structural organisation l Dehydration: replace the water in the tissue by alcohol Clearing: replace the alcohol by xylene Embedding: replace the xylene w/ melted paraffin Sectioning & mounting

B. Frozen section: Better for preserving chemical components (e.g. enzymes) Freezingcryotomystaining C. the others: Smear preparations: for blood etc; Grind preparations: for bone

2. Staining

Purpose: To make tissue section pigment for observation. H-E Staining: Hematoxylin: basic dye, purple-blue Eosin: acid dye, pink color Basophilic: components bonded by basic dye (H); pruple-bluenuclear chromatin & basophilic substance in cytoplasm Acidophilic: components bonded by acidic dye (E); pink (cytoplasm & collagenous fiber)

Neutrophilic: do not stain w/ both basic and acid dyes Metachromasia: a dye stains tissue a different color from that of dye solution, e.g. toluidine blue stains mast cells in purple color

Special staining: Argyrophilia Fluorescent staining

Silver staining of the neuron and the bile canaliculi

HO (Hoechst 33258)-PI staining shows the apoptosis in human HL-60 cells

II. Electron Microscopy

1. Transmission Electron Microscope (TEM) Using a beam of electrons (short wave-lengths) instead of visible light. Section preparation: similar to those for L.M mainly, plastic instead of paraffin, 50-70nm thick, heavy metal salts instead of HE. Resolution: 0.1-0.5nm (0.2nm) Ultrastructrue: The structure in EM Electron-dense / electron-lucent

Transmission Electron Microscope

Diagrams of TEM

2. Scanning Electron Microscope (SEM) Sowing the 3 dimensional surface Architecture of cells and tissues Resolution: 5nm

III. Histochemistry & Cytochemistry

Reveal the chemical composition in situ (e.g. proteins, a.a., nucleic acid, lipids, enzymes etc.) w/ chemical, biochemical methods. The product of chemical reaction should be insoluble / colored / electron- scattering, & be seen in LM or EM

For instance:
PAS(Periodic Acid

Schiff) reaction: for manifesting polysaccharide and proteoglycan (e.g. glycogen).

polysaccharide + HIO4
(hydroxyl group) (oxidise) Aldehyde group + Shiffs reagent (colorless) Purplish red depositor

PAS reaction in the hepatocytes

Immunocytochemistry
Based on antigen binds to specific antibody. Tissue section w/ Antigen + labelled antibody labelled Ag-Ab complex
Fluorescein labelling enzyme labelling colloidal gold labelling

NOS & GnRH positive Neurons in hypothalamus of rat

NOS positive neuron in hypothalamus of rat

m-ATPase activity in skeletal muscles

IV. tissue culture V. isotopic tracing VI. in situ hybridization (ISH)

nucleic acid molecular hybridization

Use nucleotide probe to check target fragment of intracellular DNA or mRNA in situ, in order to study the gene expression.

Expression of PSA and PSAmRNA in human prostate (histochemistry & ISH)

The method in learning histology

Combination of

the 2 dimentional structure

with 3 dimentional

Combination

of the theory with

practice
Combination

of the structure

with function
Concern

the dynamic change