UCLA Part 1) Introduction To Histological Tissues

Introduction to
Histological Tissues
Elena Stark, MD, PhD

Stephen Schettler, PhD
Department of Pathology & Laboratory Medicine
David Geffen School of Medicine at UCLA

Contributor: Paul Frank, DGSOM Class of 2014
THIS FILE HAS COPYRIGHTED MATERIAL. IT IS INTENDED FOR YOUR USE

ONLY. DO NOT DISTRIBUTE OR SHARE IT.
Non-graphical content and organization Copyright 2012 M.E. Stark, MD, PhD.
1
Main Menu
3 1. Introduction to Histological Tissues – Basic Cell Histology

4 2. Microscopy – Introduction
6 2.1 Microscopy – Light Microscopy Tissue Preparation
9 2.1.1 Microscopy – Light Microscopy Tissue Preparation – Slicing
10 2.1.2 Microscopy – Light Microscopy Tissue Preparation – Staining
12 2.1.3
Microscopy – Light Microscopy Tissue Preparation – Frozen Sections
13 3. Tissue Types
14 3.1 Tissue Types – Muscular
15 3.2 Tissue Types – Nervous
16 3.3 Tissue Types – Connective
17 3.4 Tissue Types – Epithelial
18 3.5 Tissue Types – Summary
19 4. Image Sources
The “Back to Main Menu” buttons on pages 21 to 28 have been disabled (these are the hyperlink/highlight pages).
To go to the Main Menu: first, go back to the original page by clicking BACK or ;
second, use the “Back to Main Menu” buttons on pages 3 to 20 (as they are functional).
2
1. Introduction to Histological Tissues – Basic Cell Histology
Please review the fundamentals of the cell on your own. Before you proceed with this
module make sure you are familiar with the terms listed below. We recommend
the following textbooks to learn the basics of the cell:
Young B, et al. Wheater's Functional Histology. 5th ed. Churchill Livingstone, Elsevier
Limited; March 14, 2006.
Mescher AL. Junqueira’s Basic Histology. 12th ed. McGraw-Hill Medical; August 28,
2009.
Nucleus Plasma membrane Mitochondria
Nucleolus Cytoplasm Ribosome
Chromatin Cytosol Endoplasmic reticulum
Cytoskeleton Golgi complex
Lysosomes
Peroxisomes
Secretory granule
Inclusions
3
2. Microscopy – Introduction
In the next several slides, we will provide a brief introduction

to the subject of microscopy (including the concepts of slicing
tissue and staining tissue). Please do not memorize this
information, just read through it, it’s only FYI.
There are 3 main modalities of microscopy used to study tissues:
1- Light microscopy (LM) that includes several types, such as bright-field,

fluorescence, phase-contrast, and confocal and polarizing microscopy (however,
we do NOT expect you to know the difference between these subtypes).
2- Electron microscopy (EM) that includes transmission electron microscopy (TEM)

and scanning electron microscopy (SEM).
3- Scanning probe microscopy.
Note: our modules use images taken with the light microscope (LM) and the
electron microscope (EM), so we will focus on these types in general (1 and 2
above).
4
2. Microscopy – Introduction
The main difference between light microscopy (LM) and electron microscopy (EM) is their
capacity to reveal detail.
LM can magnify up to 1000 times (x1000); while EM can magnify up to 100,000 times
(x100 000).
Electron microscopy (EM) includes 2 subtypes

• Transmission electron microscopy (TEM), also referred to simply as EM,
is an electron beam that goes through the structure to form an image.
This requires ultrathin slices of the structure that is under observation.
The result is a two-dimensional (2-D) image.
• Scanning electron microscopy (SEM) produces three-dimensional (3-D) images
of a structure’s surface.
Ignore the i.ds in the images.
LM image of 2 villi in the intestinal wall (x150). SEM image of villi in the intestinal wall (x100). TEM image of the surface of a villus
© Elsevier. Young et al. Wheater’s Functional Histology 5e – www.studentconsult.com in the intestinal wall (x56 000).
5
2.1 Microscopy – Light Microscopy Tissue Preparation
Since we will use LM images most often,

our focus will be on sample preparation for LM.
For study through LM, samples of tissues are

obtained. Biopsies (small pieces of tissue) are
taken from organs. The sample must be sliced
and stained.
The biopsy is immediately placed in a tissue

cartridge (also called a cassette); which is then
immersed in a fixative, such as formaldehyde or
glutaraldehyde.
The fixative helps:
– Arrest cell metabolism Tissue cartridges/tissue cassettes.

– Prevent enzymatic degradation
– Harden the tissue
6
Once the sample is fixed, it is then infiltrated with an embedding medium**

(usually paraffin wax or plastic resin) in order to be cut into very thin sections,
and then mounted onto glass slides.
**Most tissues must be dehydrated between fixation and embedding.
This example shows paraffin-embedded tumor blocks:

portions of a tumor have been enclosed in paraffin wax.
This can be done with any tissue, NOT just tumor tissue.
7
So far, here are the steps of tissue preparation for LM:
1. Fixation
2. Dehydration (needed in most cases)
3. Embedding
(4.) Now, the sample is ready to be sectioned into very thin slices.
(5.) Next, the slices are stained to visualize the tissue components.
The process is as follows:
Fixation > Dehydration > Embedding > Sectioning > Staining > Microscope ready
These steps sometimes result in some distortions in the cell structure or the architecture
of the tissue sample. We will see examples of this during the course.
In the next slides, we will present more information on those last 2 very important steps:
sectioning and staining.
8
2.1.1 Microscopy – Light Microscopy Tissue Preparation – Slicing
These figures will help you visualize the way three-dimensional (3-D) structures
(that have been sliced) appear in two-dimensional (2-D) histological slides.
This is a very simple concept, but essential in understanding histology.
Note: we will look at slides during lab to review this concept.
9
2.1.2 Microscopy – Light Microscopy Tissue Preparation – Staining
After being sliced, tissues are stained in order to visualize their A

components.
There are several stains for observation with the light microscope (LM)
that are commonly used:
 H&E (hematoxylin and eosin) (image A, at the top)

• Stains acidic structures purple/blue (nuclei, ribosomes, and RER).
• Stains basic structures pink (most cytoplasmic proteins).
B
 Masson’s trichrome (image B, in the middle)
• Stains extracellular structures blue (e.g. collagen).
 Giemsa (image C, at the bottom)

• Stains blood cells – nuclei stain blue/violet, cytoplasm stains light
bluish, red cells stain pale pink.
There are many other types of stains like these that we will discuss
throughout the course. C
However, in tissue preparation for observation with an electron
microscope (EM), there are NO common colored stains (like the ones
listed above). Instead, the appearance is black and white with shades
of gray (refer to the example shown on slide # 5, “2. Microscopy –
Introduction”). Other techniques are applied to prepare tissue for EM
Reference: medscape.com
(see next slide).
10
2.1.2 Microscopy – Light Microscopy Tissue Preparation – Staining
Histochemical techniques are applied to stain specific

components of cells and tissues.
A
• In sample preparation for both LM and EM.
• Particularly helpful in understanding tissue

function – often used for diagnosis of
pathological tissues.
Periodic acid–Schiff reaction (PAS) is an example

where complex carbohydrates, mucus, and glycogen
stain in deep red/magenta color (image A, at the top). Reference: www.urmc.rochester.edu
Immunohistological techniques are applied for the

identification of specific cell structures – by B
conjugating antibodies to a fluorescent stain that
binds to specific antigens (image B, at the bottom).
• In sample preparation for both LM and EM.
• Important for diagnosis and research.
Again, these descriptions are very simplistic.

We will elaborate on staining techniques as the course progresses. Reference: www.hsc.wvu.edu
11
2.1.3 Microscopy – Light Microscopy Tissue Preparation – Frozen Sections
In situations where an urgent diagnosis is needed (e.g.

when the patient is undergoing surgery and a biopsy has
been taken, the surgeon needs to know the results right
away in order to proceed), the tissue is biopsied and then
rapidly frozen to -150ºC to -170ºC by immersion in liquid
nitrogen.
Thin sections are then cut in a refrigerated chamber
(called a cryostat) and stained without previous fixation,
dehydration, or embedding.
This way of tissue preservation is NOT as good as when Nonfrozen section.
using the regular method (previously described).
Frozen sections are usually stained with H&E,
but can also be treated with histochemical and
immunohistological techniques.
Frozen section.
12
3. Tissue Types
In the next few slides, we will mention the 4 basic types of tissue that are present in the human
body. We will elaborate on these tissues throughout the year.
It is important that you understand the basic differences between tissues.
If you find this introduction too simplistic, please refer to these 2 textbooks:
Junqueira's Basic Histology by Mescher AL. 12th edition

978-0-07-163020-7
Wheater's Functional Histology by Young B, et al. 5th edition

978-0-443-06850-8
Cells in the human body are organized into tissues.

There are 4 types of tissues:
1. Muscular
2. Nervous
3. Connective
4. Epithelial
Tissues have CELLS and EXTRACELLULAR MATRIX or material (ECM) surrounding the cells.
The proportion of these 2 components varies depending on the type of tissue:
– Muscular tissue has a high cell density;

– Nervous tissue is formed almost exclusively of cells and has virtually NO extracellular material;
– Connective tissue is very rich in extracellular material and has a low proportion of cells;
– Epithelial tissue has a very high proportion of cells and very limited amount of extracellular material.
13
3.1 Tissue Types – Muscular
There are 3 types of muscle tissue: A

1. Skeletal (image A, at the top)
2. Smooth (image B, in the middle)
3. Cardiac (image C, at the bottom)
Muscle cells specialize in contraction.
Muscle tissues have high cell density and little ECM.
Muscle cell (1 in each type of muscle) B
ECM in muscle tissue
Note: we will talk more about muscle in Block 2, where we
cover the heart (cardiac muscle) and blood vessels (smooth
muscle); and also in Block 4, where we cover skeletal or
voluntary muscle (muscles we can contract at will).
ORIENT YOURSELF: Images A, B, and C are histological

slides of muscular tissue that have all been stained for LM C
observation at medium magnification.
• Image A: thin slices of voluntary muscle
• Image B: thin slices of the wall of an organ
• Image C: thin slices of the heart
(To train your eye to the differences between tissues, we will practice with
many slides in lab.) For now, focus on the general concepts.
14
3.2 Tissue Types – Nervous
Nervous cells specialize in generation and conduction of electrical impulses.

(We will cover nervous tissue in detail in Block 5.)
Nervous tissue has abundant cells of different types and virtually NO ECM.
For our purposes (before Block 5), we will identify nerves in terms of the structure highlighted
below. This is a small peripheral nerve showing the long neuronal axons, as they
appear in a cross-section. It looks like a bundle of cross-sected wires.
In this particular section, some of the axons are running obliquely – appearing oval in
shape (vs. perfect circles).
An organ showing several types of cells and tissues.

The purpose of this slide is to start becoming familiar with the appearance of a cross-sected peripheral nerve in an organ.
15
3.3 Tissue Types – Connective
Connective tissue has much ECM and few cells .

Thus, connective tissues are good “supporters” (since
ECM is usually stronger than cells). They are found
mainly forming the framework of organs or supporting
other tissues.
This week’s lab will cover connective tissue in detail.
16
3.4 Tissue Types – Epithelial
Epithelia have many cells and little ECM .

Epithelia are found mainly covering and lining organs.
This week’s lab will cover epithelia in detail.
Ignore the i.ds in the image.
17
3.5 Tissue Types – Summary
Extracellular matrix
Tissue type Cells Function
(ECM)
• Movement of body (skeletal)
• Contractile cells • Very small • Involuntary movement (smooth)
Muscular
amount • Beating of heart (cardiac)
• Neurons containing long

processes, called axons • Virtually NONE • Conduction of nerve impulses
Nervous and dendrites
• Supporting cells
• Fibroblasts • Large amount • Support of organs

Connective • Other cells • Protection of organs
• Tightly-packed cells • Very small • Lining and covering surfaces of

Epithelial with mostly flat edges amount body and body cavities
• Glands: secretion (to be discussed
in the next module: “Epithelial Tissue”)
18
4. Image Sources
1. Young B, et al. Wheater's Functional Histology. 5th ed.

Churchill Livingstone, Elsevier Limited; March 14, 2006.
2. Mescher AL. Junqueira’s Basic Histology. 12th ed.
McGraw-Hill Medical; August 28, 2009.
3. www.medscape.com
4. www.urmc.rochester.edu
5. www.hsc.wvu.edu
6. UCLA David Geffen School of Medicine, Integrative Anatomy.
19
20
BACK 3.1 Tissue Types – Muscular


21
BACK 3.1 Tissue Types – Muscular


22
BACK 3.2 Tissue Types – Nervous


23
BACK 3.2 Tissue Types – Nervous


24
BACK 3.3 Tissue Types – Connective

other tissues.
25
BACK 3.3 Tissue Types – Connective

other tissues.
26
BACK 3.4 Tissue Types – Epithelial

27
BACK 3.4 Tissue Types – Epithelial

28

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Introduction to

Elena Stark, MD, PhD

THIS FILE HAS COPYRIGHTED MATERIAL. IT IS INTENDED FOR YOUR USE

3 1. Introduction to Histological Tissues – Basic Cell Histology

Nucleus Plasma membrane Mitochondria

Nucleolus Cytoplasm Ribosome

Chromatin Cytosol Endoplasmic reticulum

Cytoskeleton Golgi complex

In the next several slides, we will provide a brief introduction

There are 3 main modalities of microscopy used to study tissues:

1- Light microscopy (LM) that includes several types, such as bright-field,

2- Electron microscopy (EM) that includes transmission electron microscopy (TEM)

3- Scanning probe microscopy.

Electron microscopy (EM) includes 2 subtypes

Since we will use LM images most often,

For study through LM, samples of tissues are

The biopsy is immediately placed in a tissue

The fixative helps:

– Arrest cell metabolism Tissue cartridges/tissue cassettes.

Once the sample is fixed, it is then infiltrated with an embedding medium**

This example shows paraffin-embedded tumor blocks:

So far, here are the steps of tissue preparation for LM:

The process is as follows:

Note: we will look at slides during lab to review this concept.

After being sliced, tissues are stained in order to visualize their A

 H&E (hematoxylin and eosin) (image A, at the top)

 Giemsa (image C, at the bottom)

Histochemical techniques are applied to stain specific

• Particularly helpful in understanding tissue

Periodic acid–Schiff reaction (PAS) is an example

Immunohistological techniques are applied for the

• In sample preparation for both LM and EM.

• Important for diagnosis and research.

Again, these descriptions are very simplistic.

In situations where an urgent diagnosis is needed (e.g.

Junqueira's Basic Histology by Mescher AL. 12th edition

Wheater's Functional Histology by Young B, et al. 5th edition

Cells in the human body are organized into tissues.

– Muscular tissue has a high cell density;

There are 3 types of muscle tissue: A

ORIENT YOURSELF: Images A, B, and C are histological

Nervous cells specialize in generation and conduction of electrical impulses.

An organ showing several types of cells and tissues.

Connective tissue has much ECM and few cells .

Epithelia have many cells and little ECM .

Ignore the i.ds in the image.

• Neurons containing long

• Fibroblasts • Large amount • Support of organs

• Tightly-packed cells • Very small • Lining and covering surfaces of

1. Young B, et al. Wheater's Functional Histology. 5th ed.

There are 3 types of muscle tissue: A

ORIENT YOURSELF: Images A, B, and C are histological

There are 3 types of muscle tissue: A

ORIENT YOURSELF: Images A, B, and C are histological

Nervous cells specialize in generation and conduction of electrical impulses.

An organ showing several types of cells and tissues.

Nervous cells specialize in generation and conduction of electrical impulses.

An organ showing several types of cells and tissues.

Connective tissue has much ECM and few cells .

Connective tissue has much ECM and few cells .

Epithelia have many cells and little ECM .

Ignore the i.ds in the image.

Epithelia have many cells and little ECM .