Histology and Embryology Website：m-learning.zju.edu.cn Histology and Embryology LiDongMei may75119@163.com

Reference books 1. Jun min T, Ji cheng L. Textbook of Histology and Embryology. Peking University Medical Press, 2011 2. William k. Ovalle & Patrick C. Nahirney. Netter’s essential histology, Elsevier Health Sciences,2007 3. Gaetner MP, Hlatt JL. Colour Textbook of Histology. Williams & Wilkins, 2007

Course web http://m-learning.zju.edu.cn (2018 Histology and Embryology)

The composition of final score: Attendance(including Lecture and lab)：10% Lab Quiz: 15% （each quiz 5%） 3. Lab test: 25% 4. Final written examination：45% Histology： 35-40% Embryology：10-15% If you want to pass this course: final written examination score must be more than 50 and final score>60

Preparation of laboratory work Have the tools ready: Pencils (red-blue pencil) pen, rubber, ruler and so on Guide book and homework(lab teacher hand out in the first lesson)

Chapter 1 Introduction

Ⅰ.   What’s histology? What’s Embryology? Ⅱ.   Why we study it ? Ⅲ.   How to study it ? Ⅳ.  Histological methods

I. What’s histology? Histology (Greek words): /histo-tissue /logia-study of ,or knowledge of Histology means the knowledge of tissue, is a branch of Anatomy.

Anatomy: ---gross anatomy ---microscopic anatomy/microanatomy Structures related to function. Histology is a science which study the microstructure and the relationship between the structure and function of human being.

What we will learn in this course Cell Tissue: 4 basic tissues Organ and system

Cell: smallest unit of structure and function of body ↓ tissue: group of cell and extracellular ground substance four basic tissue: ---epithelium ↓ ---connective tissue ---muscular tissue ---nervous tissue organ: made up of tissue, have special shape, structure and function system: organs which have related function get together.

I.  What’s Embryology? Embryology is a kind of science which study the processes and the regulations of the development of human fetus. 1.Preembryonic period: 1-2W 2.Embryonic period: 3-8W 3.Fetal period:9-38W

II. Why we study it ? To recognize normal tissue and cells To acquire basic skill which you will use throughout your career!!!

III. How to study it ? What you need to do Look Think Compare Remember Idealized images

Ⅳ. Histological methods ---Development of histology depends on the development of technique. ---Histology studies the microstructures. So, we should have the aid of microscope to study. Several types of microscopes are available.

Some Instruments for Histologcal Research

Light Microscope（LM） 1. Basic methods of histology ——Observation of histological slides under LM Light Microscope（LM）

Preparation of tissue for LM The most routine one is paraffin section stained with hematoxylin and eosin(H&E) The steps: （1）Obtaining the specimen: fresh, small pieces ( less than 5mm3)-tissue block （2） Fixation: use formalin or Bouin’s to preserve structural organisation （3）Dehydration: use ethyl alcohol to get rid of water of tissue and cell

（4） Clearing: use xylene to get rid of alcohol *alcohol and xylene are embedding mediums （5）Embedding: firstly, heat the paraffin, make it melt, then put tissue block into melted paraffin, allow paraffin harden, the tissue block is embedded in.

（6）Sectioning: use microtome to cut the tissue into 3~8um thick sections, then mounted them on glass slides

（7） H&E staining ---Hematoxylin: basic stain, combines with acidic components, make them appear blue colour- basophilic, i.e. cell nucleus, hyaline cartilage ---Eosin: acidic stain, combines with basic components, make them appear pink colour- acidophilic (eosinophilic), i.e. cytoplasm H&E is the most commonly method in histological study.

Stain methods H.E 伊 红 苏木精 Eosin Hematoxylin Acidic stucture： Cytoplasma lysosome、Mit.fibles Basic stucture： Nucleus RER Free ribosome

Hematoxylin Blue Eosin Pink acidophilic (eosinophilic) basophilic Hematoxylin Eosin H&E

Electron microscope Transmission electron microscope (TEM) Scanning electron microscope(SEM)

TEM

Basophilic granulocyte (TEM) 在血涂片上很难找到…因为数量少。 C核呈“S”形，染色浅，轮廓不清楚。 胞质含有许多大小不等，分布不均的紫蓝色的嗜碱性颗粒。 EM：大量的嗜碱性颗粒，圆形椭圆形。 颗粒内有时可见板层状结构。 Basophilic granulocyte (LM)

Preparation of tissue for EM The steps are same to preparation for LM a.   tissue block: more small, less than 1mm3 b.   plastic materials for embedding c.  ultra-thin sections is about 30-50nm thick( use ultramicrotome) d.   heavy metal salts- increase staining contrast ---lead citrate ---uranyl acatate

SEM

Some methods for Histological Research

Histochemistry & Cytochemistry To combine histological and cytological methods with chemical and biochemical methods and reveal the chemical composition of tissue and cell in situ.

Immunohistochemistry To use labelled antibodies as specific reagents for localising tissue and cell constituents (antigens) in situ

In Situ Hybridization incubate tissues with probe to detect cells expressing gene

Questions Give the names of four basic tissues？ What does basophilic and acidophilic mean?