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Boys and girls Welcome. About practical lessons. 1 Lab: morphology lab 5 and 6 On fourth floor of east building class one in lab5, class two in lab6 2 Preparation: Textbooks laboratory manual practical paper and so on 3 no food in lab. Exercises after class.

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Boys and girls

Welcome


About practical lessons

1 Lab: morphology lab 5 and 6

On fourth floor of east building

class one in lab5,

class two in lab6

2 Preparation:

Textbooks

laboratory manual

practical paper and so on

3 no food in lab


Exercises after class

1 HE staining

Acidophilc,

Basophilc

2 Features of electron microscopy

Tissue slide in practical lessons:

Spinal ganglion


Self Introduction

Zhao Hongxian

Lecturer

Master graduate

Email:[email protected]

QQ:15673656


Faculty of

the department of histology and embryology


第一章 组织学绪论

Chapter 1 Histological Introduction


2 Significants(why)

3 Technology(how)

Main contents:

1 Definiton、contents

(what)


1 Definiton,study contents

1.1 Definiton of histology(组织学)

Histo = tissue, logos = study or science

A science which studies

the normalmicrostructures of the body

and the basic functions.


1.2 Study contents

Cell(细胞):

Basic unit of structure and function of body

Tissue(组织):

★Definition: made of cells

and extracellular matrix

★Types:(1) Epithelial tissue,Epi

(2) Connective tissue, CT

(3) Muscle tissue

(4) Nervous tissue

3. Organ(器官)and system(系统)


Differences?


2 Significants

Basic science:WHY?

Precondition of studying other medical disciplines, especially physiology

and pathology


Intern (exercitation)

Graduate

Clinic (Medicine, Surgery, Gynecology, Pedology, etc.)

Pathphysiology

Pharmocology

Basic medicine &Surgery

Biochemistry

Physiology

Microbiology

Immunology

Pathology

Parasitology

macrostructure

microscope

First

Year

Histology & Embryology

Anatomy


3 Histological techniques

3.1 Observing tools(your weapon)

光镜(light microscope, LM)

电镜(electron microscope, EM)

3.2 Samples

paraffin section, frozen section,

ultrathin section ,Smear, stretched preparation,

ground section, living cell.

3.3 Treatments of sample

HE staining, special staining,, histochemistry, in situ hybridization, heavy metal staining, autoradiography vital staining


3.1 observing tools

3.1.1 Light microscope

Based on the interaction of

light and tissue components

Compositions:

mechanical and optical parts


3.1.2 Electron microscope

Transmission electron microscope (TEM)

Scanning electron microscope (SEM)

SEM

TEM


TEM


Transmission electron microscope

(1)Features:★

① beam of electrons

② Electromagnetic lenses

③ Ultrathin (40-90 nm )

④ Heavy metal staining

(Uranyl acetate, Lead Citrate, lead nitrate)

(2)Dark areas of an electron micrograph are usually

called electron dense ★,

Light areas are called electronlucent★


Scanning electron microscope

Do not pass through the specimen,interact with surface of the specimen,

and produce reflected or emitted electrons→ → captured by a detector→ → transmit them to amplifiers and other devices→ → projected into a monitor→resulting in a pseudo-three-dimensional black-and-white image

SEM → →Pseudo-three-dimensional ,surface structures

TEM → →Plane, intra structures


3.2 Samples

3.2.1 Paraffin section(石蜡切片):

Classic and main sample

Steps of making paraffin section:For observing clearly

① Obtainthe sample

② Fix

③ Dehydrate, Clear

④ Embed

⑤ Section (5-10 um )

⑥ Stain(increase contrast)

⑦ Mount and label


3.2.2 0ther samples(self-study):

Frozen section

Ultrathin section

Smear

Stretched preparation

Ground section

Living cell


Smear

ground section

stretched preparation


3 Treatments of samples

Treatments

AUSAS

3.1 H E staining:★

① The most commonly used

② H (hematoxylin),blue basic dye

make acid substance blue

③ E (eosin),red acidic dye

make basic substance red

④ Combination of hematoxylin and eosin

is called HE staining


3.1.1 Terms relating to HE staining

a.★ Tissue components with an high affinity for basic dyes are termed basophilic

b.★ Tissue components with an high affinity for acid dyes are termed acidophilic

(Gr. Phileo means love)


3.1.2 Rules of HE staining

A. Nucleus is generally stainedblue or purple

(why);

The more dark the nucleus stains, the more inactive the cell functions

B. Cytoplasm is generally stainedred(why);

RERs or ribosomes are stained blue in cytoplasm, which indicates cells actively synthesis proteins


3.2 Histochemistry

3.2.1 General histochemistry

1)Principles:★

target substances+A reagent→→insoluble colored or electron-dense compounds →→localization of target substances by means of light or electron microscopy

2)Localizing:

Ions

Polysaccharides & Oligosaccharides:PAS reaction

Lipids

Nucleic acids:Feulgen reaction

Proteins (enzymes)


3.2.2 Immunohistochemistry

1)Princple:Specific affinity between antigen(target protein or peptide) and antibody

Lables:

Fluorescent compound,peroxidase,

gold particle


in situ hybridization

3.2.3 In situ hybridization

1)Probe: A known segment of single-stranded DNA or RNA that is complementary to the target nucleic acid. probe must be tagged with a lable.

2 ) Principle:Because of complementary→ probe+target nucleic acid→hybridizing→→detecting the target nucleic acid(DNA,RNA)


3.2.4 Other treatments of samples(self-study)

Special staining,

Vital staining

Heavy metal staining,

Autoradiography


4 About studying histology

1) preparation,lectures and review a lesson

2) systematic study:

Syetems, organs,tissues,cells, organelles

3) pay more attention to study method

4) asking and thinking

5) two dimentional and three dimentional

6) various sections of one structure

7) artifacts in tissue samples


Cell Membrane

Cytoplasm

Nucleus


obtain、fix

immerse

Dehydrate,clear

embed


section


stain

mount

lable


rules


Sliver staining


stretched preparation :demonstrating macrophage


Ground section of bone


This peripheral blood smear is stained with the Wright's stain

Blood smear, Giemsa stain.


PCNA阳性表达情况,免疫细胞化学染色,×400


免疫组织化学(荧光素标记)

毛细血管内皮细胞呈vWF阳性


OVER

TANAKS A LOT


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