Part 1 - Get a Lab Appointment and Install Software:
Set up an Account on the Scheduler (FIRST TIME USING NANSLO):
Find the email from your instructor with the URL (link) to sign up at the scheduler.
Set up your scheduling system account and schedule your lab appointment.
NOTE: You cannot make an appointment until two weeks prior to the start date of this lab assignment.
You can get your username and password from your email to schedule within this time frame.
Install the Citrix software: – go to http://receiver.citrix.com and click
download > accept > run > install (FIRST TIME USING NANSLO).
You only have to do this ONCE. Do NOT open it after installing. It will work automatically when you go
to your lab. (more info at
http://www.wiche.edu/info/nanslo/creative_science/Installing_Citrix_Receiver_Program.pdf)
Scheduling Additional Lab Appointments:
Get your scheduler account username and password from your email.
Go to the URL (link) given to you by your instructor and set up your appointment.
(more info at http://www.wiche.edu/nanslo/creative-science-solutions/students-scheduling-labs)
Changing Your Scheduled Lab Appointment:
Get your scheduler account username and password from your email. Go to http://scheduler.nanslo.org
and select the “I am a student” button. Log in to go to the student dashboard and modify your
appointment time. (more info at http://www.wiche.edu/nanslo/creative-science-solutions/studentsscheduling-labs)
Part 2 – Before Lab Day:
Read your lab experiment background and procedure below, pages 1-24.
Submit your completed Pre-Lab 1-5 Questions (pages 5-8) per your faculty’s instructions.
Watch the Microscope Control Panel Video Tutorial
http://www.wiche.edu/nanslo/lab-tutorials#microscope
Part 3 – Lab Day
Log in to your lab session – 2 options:
1)Retrieve your email from the scheduler with your appointment info or
2) Log in to the student dashboard and join your session by going to http://scheduler.nanslo.org
NOTE: You cannot log in to your session before the date and start time of your appointment. Use
Internet Explorer or Firefox.
Click the yellow button on the bottom of the screen and follow the instructions to talk to your lab
partners and the lab tech.
Remote Lab Activity
SUBJECT SEMESTER: ____________
TITLE OF LAB: Histology - Connective Tissue
Lab format: This lab is a remote lab activity.
Relationship to theory (if appropriate): In this lab you will learn the underlying principles
behind the histological study of tissues.
Instructions for Instructors: This protocol is written under an open source CC BY license. You
may use the procedure as is or modify as necessary for your class. Be sure to let your students
know if they should complete optional exercises in this lab procedure as lab technicians will not
know if you want your students to complete optional exercises.
Instructions for Students: Read the complete laboratory procedure before coming to lab.
Under the experimental sections, complete all pre-lab materials before logging on to the
remote lab. Complete data collection sections during your online period, and answer questions
in analysis sections after your online period. Your instructor will let you know if you are
required to complete any optional exercises in this lab.
Remote Resources: Primary – Microscope, Secondary – Histology slide set.
CONTENTS FOR THIS NANSLO LAB ACTIVITY:
Learning Objectives...............................................................................................
Background Information ......................................................................................
Equipment ............................................................................................................
Preparing for this NANSLO Lab Activity ...............................................................
Experimental Procedure ......................................................................................
Pre-lab Exercise 1: Areolar Connective Tissue ....................................................
Exercise 1: Areolar Connective Tissue .................................................................
Pre-lab Exercise 2: Andispose Connective Tissue ...............................................
Exercise 2: Andispose Connective Tissue ............................................................
Pre-lab Exercise 3: Reticular Connective Tissue .................................................
Exercise 3: Reticular Connective Tissue .............................................................
Pre-lab Exercise 4: Dense Connective Tissue ......................................................
Exercise 4: Dense Connective Tissue ..................................................................
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CONTENTS FOR THIS NANSLO LAB ACTIVITY – CONT’D
Pre-lab Exercise 5: Ground Substance Based Connective Tissue .......................
Exercise 5: Ground Substance Based Connective Tissue ....................................
Summary Questions .............................................................................................
Creative Commons Licensing ...............................................................................
U.S. Department of Labor Information ................................................................
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LEARNING OBJECTIVES:

Overview of histology and tissue types
1. Define the term histology.
2. List four major tissue types.
3. Contrast the general features of the four major tissue types.

Microscopic anatomy, location, and functional roles of epithelial tissue
1. Classify the different types of connective tissue based on distinguishing
structural characteristics.
2. Describe locations in the body where each type of connective tissue can be
found.
3. Describe the functions of each type of connective tissue in the human body and
correlate functions with structure for each tissue type.
4. Identify the different types of connective tissue using proper microscope
technique.
BACKGROUND INFORMATION:
A living organism is composed of a variety of cells of different sizes, shapes, structures and
specialized functions. Cells of similar type are usually organized into groups. A group of cells
with similar size, shape, structure and function form a tissue. There are four general classes of
tissues. These classes are epithelial, connective, muscle, and neuronal. In this lab, we will use
histology to examine several connective tissues.
Histology1 is the branch of biology concerned with the composition and structure of plant and
animal tissues in relation to their specialized functions. The terms histology and microscopic
anatomy are sometimes used interchangeably, but a fine distinction can be drawn between the
two studies. The fundamental aim of histology is to determine how tissues are organized at all
structural levels from cells and intercellular substances to organs.
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Connective tissues maintain the form of the body and its organs and provide cohesion and
internal support. The connective tissues include several types of fibrous tissue that vary only in
their density and cellularity as well as the more specialized and recognizable variants — bone,
ligaments, tendons, cartilage, and adipose (fat) tissue.
Some organs are suspended from the wall of a body cavity by thin sheets of connective tissue
called mesenteries. Others are embedded in adipose tissue, a form of connective tissue in
which the cells are specialized for the synthesis and storage of energy-rich reserves of fat or
lipid. The entire body is supported from within by a skeleton composed of bone, a type of
connective tissue endowed with great resistance to stress owing to its highly ordered laminated
structure and to its hardness, which results from deposition of mineral salts in its fibers and
amorphous matrix. The individual bones of the skeleton are held firmly together by ligaments,
and muscles are attached to bone by tendons, both of which are examples of dense connective
tissue in which many fiber bundles are associated in parallel array to provide great tensile
strength. At joints, the articular surfaces of the bones are covered with cartilage, a connective
tissue with an abundant intercellular substance that gives it a firm consistency well adapted to
permitting smooth gliding movements between the apposed surfaces. The synovial membrane
which lines the margins of the joint cavity and lubricates and nourishes the joint surfaces is also
a form of connective tissue.
Components of the connective tissue: All forms of connective tissue are composed of (1)
extracellular fibers, (2) an amorphous matrix called ground substance, and (3) cells (stationary
and migrating. We will discuss each component below. The proportions of these components
vary from one part of the body to another depending on the local structural requirements. The
anatomical classification of the various types of connective tissue is based largely upon the
relative abundance and arrangement of these components.
Extracellular fibers: The fibrous component of connective tissue fall into three kinds:
collagenous, elastic, and reticular. The collagenous fibers are the most abundant and are
composed of the protein collagen which is of commercial and medical interest. The behavior of
collagen varies tremendously based on how it is treated. Leather is the collagen-dense dermis
of animal skins preserved and toughened by the process called tanning. Fresh collagen dissolves
in hot water and produces gelatin. Under appropriate conditions, collagen can be brought into
solution without chemical change. The fundamental units in such solutions are slender
tropocollagen. Collagen appears to be secreted in this form by the connective-tissue cells called
fibroblasts, and the tropocollagen molecules assemble extracellularly to form striated collagen
fibrils.
Elastic fibers are composed of the protein elastin and differ from collagenous fibers in
dimension, pattern, and chemical composition. They do not have uniform subunits comparable
to the unit fibrils of collagen. They present a variable appearance in electron micrographs.
Sometimes they appear to have an amorphous core surrounded by minute fibrils, while in other
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sites they appear to consist exclusively of dense amorphous material. Whether there are in fact
two components or whether these are differing forms of the same substance is not yet clear. At
the light-microscope level, the fibers vary in diameter and often branch and reunite to form
extensive networks in loose connective tissue. When present in high concentration, they
impart a yellow color to the tissue. In elastic ligaments, the fibers are very coarse and are
arranged in parallel bundles. In the walls of arteries, elastin is present in the form of sheets or
membranes perforated by openings of varying size. As their name implies, elastic fibers are
highly distensible and, when broken, recoil like rubber bands. Changes in this property and
diminution in their numbers are thought to be, in part, responsible for the loss of elasticity of
the skin and of the blood-vessel walls in old age.
Reticular fibers are distinguished by their tendency to form fine-meshed networks around cells
and cell groups. They are small fascicles of typical collagen fibrils interwoven to form a network.
Ground substance: The amorphous ground substance of connective tissue is a transparent
material with the properties of a viscous solution or a highly hydrated thin gel. Its principal
constituents are large carbohydrate molecules or complexes of proteins and carbohydrates. All
substances passing to and from cells must pass through the ground substance. Variations in its
composition and viscosity may therefore have an important influence on the exchange of
materials between tissue cells and the blood. Its physical consistency also constitutes a barrier
to the spread of particulates introduced into the tissues. While all types of connective tissue
have at least a small amount of ground substance, two types consist predominately of it.
Cartilages are a form of connective tissue in which the ground substance is abundant
and of a firmly gelated consistency that endows this tissue with unusual rigidity and
resistance to compression. The cells of cartilage, called chondrocytes, are isolated in
small lacunae within the matrix. Although cartilage is a vascular (without blood supply),
gaseous metabolites and nutrients can diffuse through the aqueous phase of the gel-like
matrix to reach the cells.
Bones like cartilage consist of cells, fibers, and ground substance, but, in addition, the
extracellular components are impregnated with minute crystals of calcium phosphate.
The mineralization of the matrix is responsible for the hardness of bone. Because the
hardness of bone the tissue does not diffuse nutrients rapidly, bone tissue is
vascularized. It also provides a large reserve of calcium that can be drawn upon to meet
unusual needs for this element elsewhere in the body. The structural organization of
bone is adapted to give maximal strength for its weight-bearing function with minimum
weight. The cells in bone are called osteocytes.
Cells of connective tissue include two types that are relatively stationary—fibroblasts and
adipose cells—and several types of motile migrating cells—mast cells, macrophages,
monocytes, lymphocytes, and eosinophils. All the cells of connective tissue develop during
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embryonic life from the mesenchyme, a network of primitive stellate cells that have the
potential to differentiate along several different lines depending upon local conditions.
Stationary cells: The ubiquitous fibroblasts are the principal active cells of connective
tissue, occurring as long spindle-shaped cells stretched along bundles of collagen fibrils.
Their function is to secrete collagen and constituents of the ground substance and to
maintain these extracellular tissue components.
Adipose, or fat, cells are connective-tissue cells that are specialized for the synthesis
and storage of reserve nutrients. They receive glucose and fatty acids from the blood
and convert them to lipid, which accumulates in the body of the cell as a large oil
droplet. This distends the cell and imposes upon it a spherical form. The nucleus is
displaced to the periphery, and other metabolically active constituents of the cell are
confined to a thin rim of cytoplasm around the large central droplet of lipid. Adipose
cells may occur in small numbers anywhere in connective tissue, but they tend to
develop preferentially along the course of small blood vessels where they accumulate in
such large numbers that they become the predominant cellular element.
Migrating cells: In addition to the relatively fixed cell types described above, there are free
cells that reside in the interstices of loose connective tissue. These vary in their abundance and
are free to migrate through the extracellular spaces. (Do you need to define or discuss these here
as you mentioned them????: mast cells, macrophages, monocytes, lymphocytes, and eosinophils.)
References:
1. "histology." Encyclopedia Britannica.
http://www.britannica.com/EBchecked/topic/267172/histology (22 April, 2014)
EQUIPMENT:
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Paper
Pencil/pen
Slides
o Areolar tissue slide
o Adipose tissue slide
o Lymph node slide
o Tendon white fibrous connective tissue slide
o Skin pigmented slide
o Hyaline cartilage slide
o Compact bone slide
Computer with Internet access (for the remote laboratory and for data analysis)
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PREPARING FOR THIS NANSLO LAB ACTIVITY:
Read and understand the information below before you proceed with the lab!
Scheduling an Appointment Using the NANSLO Scheduling System
Your instructor has reserved a block of time through the NANSLO Scheduling System for you to
complete this activity. For more information on how to set up a time to access this NANSLO lab
activity, see www.wiche.edu/nanslo/scheduling-software.
Students Accessing a NANSLO Lab Activity for the First Time
For those accessing a NANSLO laboratory for the first time, you may need to install software on
your computer to access the NANSLO lab activity. Use this link for detailed instructions on
steps to complete prior to accessing your assigned NANSLO lab activity –
www.wiche.edu/nanslo/lab-tutorials.
Video Tutorial for RWSL: A short video demonstrating how to use the Remote Web-based
Science Lab (RWSL) control panel for the air track can be viewed at
http://www.wiche.edu/nanslo/lab-tutorials#microscope.
NOTE: Disregard the conference number in this video tutorial.
AS SOON AS YOU CONNECT TO THE RWSL CONTROL PANEL: Click on the yellow button at the
bottom of the screen (you may need to scroll down to see it). Follow the directions on the pop
up window to join the voice conference and talk to your group and the Lab Technician.
EXPERIMENTAL PROCEDURE:
Once you have logged on to the remote lab system, you will perform the following laboratory
procedures. See Preparing for the Microscope NANSLO Lab Activity below.
During the study of each type of human tissue, you are required to photograph and label a
representative sample of each tissue type. You should begin your microscopic examination of the tissue
slide using the 4X objective to give you a perspective of the entire structure. This is very important,
since sometimes more than one tissue type is often present on each slide. All observations should be
made using the 40X or 60X objective lens (whichever gives you the best results.)
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PRE-LAB EXERCISE 1: Areolar Connective Tissue
Pre-Lab Questions:
1. Based on your course research and pre-lab readings what characteristics do you expect
to see in Areolar connective tissue?
EXERCISE 1: Areolar Connective Tissue
Data Collection:
2. Select the areolar tissue slide from the microscope interface. Using the 10X objective,
locate the tissue sample and bring it into focus.
3. Carefully work your way through all the objectives focusing with each one until you
reach the 40X or 60X objective and capture an image of the areolar connective tissue.
Insert your image below.
Analysis (can be completed offline):
4. Using your image from Exercise 1, question 3, label the 3 different parts of a cell
(nucleus, cytoplasm and cell membrane) and the different types of fibers.
5. Based on your observation, describe the function of the areolar connective tissue in the
human body and correlate its function with its structure.
6. Where do you find the areolar connective tissue inside the human body?
PRE-LAB EXERCISE 2: Andipose Connective Tissue
Pre-Lab Questions:
1. Based on your course research and pre-lab readings what characteristics do you expect
to see in adipose connective tissue?
EXERCISE 2: Andipose Connective Tissue
Data Collection:
2. Select the adipose tissue slide from the microscope interface. Using the 10X objective,
locate the tissue sample and bring it into focus.
3. Carefully work your way through all the objectives, focusing with each one, until you
reach the 40X or 60X objective and capture an image of the adipose tissue. Insert your
image below.
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Analysis (can be completed offline:)
4. Using your image from Exercise 2, question 2, Label the 3 different parts of a cell
(nucleus, cytoplasm and cell membrane) and the different types of fibers.
5. Based on your observation, describe the function of the adipose tissue in the human
body and correlate its function with its structure.
6. Based on your observation, what is a particular difference you have noticed between
the adipose tissue and the areolar tissue?
7. Where do you find the adipose tissue inside the human body?
PRE-LAB EXERCISE 3: Reticular Connective Tissue
Pre-Lab Questions:
1. Based on your course research and pre-lab readings, what characteristics do you expect
to see in reticular connective tissue?
EXERCISE 3: Reticular Connective Tissue
Data Collection:
2. Select the lymph node or the spleen slide from the microscope interface. Using the 10X
objective, locate the tissue sample and bring it into focus.
3. Carefully work your way through all the objectives, focusing with each one until you
reach the 40X or 60X objective and capture an image of the reticular connective tissue.
Insert your image below.
Analysis (can be completed offline:)
4. Using your image from Exercise 3, question 2, label the 3 different parts of a cell
(nucleus, cytoplasm and cell membrane) and the different types of fibers
5. Based on your observation, describe the function of the reticular tissue in the human
body and correlate its function with its structure.
6. Where do you find the reticular connective tissue inside the human body?
PRE-LAB EXERCISE 4: Dense Connective Tissue
Pre-Lab Questions:
1. Based on your course research and pre-lab readings, what characteristics do you expect
to see in dense regular connective tissue?
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2. Describe the differences you expect to see between dense regular and irregular
connective tissue.
EXERCISE 4: Dense Connective Tissue
Data Collection:
3. Select the tendon white fibrous connective tissue slide from the microscope interface.
Using the 10X objective, locate the tissue sample and bring it into focus.
4. Carefully work your way through all the objectives, focusing with each one until you
reach the 40X objective and capture an image of the dense regular connective tissue.
Insert your image below.
5. Select the skin pigmented slide from the microscope interface. Using the 10X objective,
locate the tissue sample and bring it into focus.
6. Carefully work your way through all the objectives, focusing with each one until you
reach the 40X or 60X objective and capture an image of the dense irregular connective
tissue. Insert your images below.
Analysis (can be completed offline):
7. Using your image from Exercise 4, question 4, label the 3 different parts of a cell
(nucleus, cytoplasm and cell membrane) and the different types of fibers.
8. Based on your observation, describe the function of the dense regular connective tissue
in the human body and correlate its function with its structure.
9. Where do you find the dense regular tissue inside the human body?
10. Using your image from Exercise 4, question 6, label the 3 different parts of a cell
(nucleus, cytoplasm and cell membrane) and the different types of fibers.
11. Based on your observation, describe the function of the dense irregular connective
tissue in the human body and correlate its function with its structure.
12. Where do you find the dense irregular tissue inside the human body?
PRE-LAB EXERCISE 5: Ground Substance Based Connective Tissue
Pre-Lab Question:
1. Based on your course research and pre-lab readings what are the similarities and
differences you expect to see between the cartilage and bone?
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EXERCISE 5: Ground Substance Based Connective Tissue
Data Collection:
2. Select the hyaline cartilage 2 from the microscope interface. Using the 10X objective,
locate the tissue sample and bring it into focus.
3. Carefully work your way through all the objectives, focusing with each one, until you
reach the 40X or 60X objective and capture an image of the hyaline cartilage. Insert
your image below.
4. Select the bone dry ground human 1 slide from the microscope interface. Using the 10X
objective, locate the tissue sample and bring it into focus.
5. Carefully work your way through all the objectives, focusing with each one, until you
reach the 40X or 60X objective and capture an image of the compact bone. Insert your
image below.
Analysis (can be completed offline):
6. Using your image from Exercise 5, question 2, label the 3 different parts of a cell
(nucleus, cytoplasm and cell membrane) and the ground substance.
7. Describe the function of the cartilage in the human body and correlate its function with
its structure.
8. Where do you find the hyaline cartilage inside the human body?
9. Using your image from Exercise 5, question 4, label the 3 different parts of a cell
(nucleus, cytoplasm and cell membrane) and the different types of fibers.
10. Based on your observation, describe the organization of the compact bone.
11. What is the function of the compact bone in the human body and correlate its function
with its structure.
12. Where do you find the compact bone inside the human body?
SUMMARY QUESTIONS:
Answer the following questions regarding the connective tissue.
1. Where is the nucleus of an adipocyte located? Why?
2. Where are the functional differences between dense irregular and dense regular
connective tissue? How do these differences help each to perform their unique
function?
3. Based on your observation, what is a particular difference you have noticed between
the dense irregular and the dense regular connective tissues?
4. Based on your observation, what is a particular difference you have noticed between
the hyaline cartilage and the compact bone?
5. Discuss the impact of diet and hormones on the onset of osteoporosis.
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For more information about NANSLO, visit www.wiche.edu/nanslo.
All material produced subject to:
Creative Commons Attribution 3.0 United States License 3
This product was funded by a grant awarded by the U.S.
Department of Labor’s Employment and Training Administration.
The product was created by the grantee and does not necessarily
reflect the official position of the U.S. Department of Labor. The
Department of Labor makes no guarantees, warranties, or
assurances of any kind, express or implied, with respect to such
information, including any information on linked sites and
including, but not limited to, accuracy of the information or its
completeness, timeliness, usefulness, adequacy, continued
availability, or ownership.
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