MODULE HUMAN HISTOLOGY

Chapter 1 - The Cell

The goal of this chapter is to learn how to look for items of interest in histological specimens using
light microscopy. A variety of cells, tissues, and organs are provided as samples.
Characteristics to notice and observe:

• Size of the cell

• Shape of the cell
• Nuclear/cytoplasmic ratio
• Chromatin condensation: heterochromatin or euchromatin
• Cytoplasmic staining: basophilic or acidophilic
• Secretion granules
• Special staining properties

Light Microscope
A microscope is an instrument used to see objects that are
too small to be seen by the naked eye.
The human eye can resolve objects separated by at least
0.250 mm. A microscope magnifies an image so smaller
details can be seen by the eye.
A light microscpe forms a magnified image by focusing light
with glass lenses on a thin tissue section.

• Condenser lens - focus the light from the illumination source

onto the specimen
• Objective lens - collects light that passes through the
specimen, enlarges and projects an image onto the eyepiece
• Eyepiece lens - further magifies images and project onto the
retina

Bright-field illumination is used with specimens stained with

colored dyes. It has a resolution of ~0.25 µm (or ~1000x
greater than the unaided eye).

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Histological Stains
Biological material is inherently of low contrast and provides little to see in a standard bright field
microscope unless treated with a histological stain.
The most widely used histological stains
differentiate between the acidic and basic
components of cells and tissues.
Basic dyes have a net positive charge and bind to
components of cells and tissues that are negatively
charged.

• Phosphate groups of nucleic acids (DNA and

RNA)
• Sulfate groups of some polysaccharides
(glycosaminoglycans) and some proteins (mucus).

Tissue components that stain with basic dyes are

referred to as basophilic.
Acidic dyes have a net negative charge and bind to components of cells and tissues that are
positively charged.
Hematoxylin and eosin (or H&E) is the most commonly used stain in histology. This stain works
well with a variety of fixatives and stains a broad range of cytoplasmic, nuclear, and extracellular
matrix features.

• Hematoxylin is a postively charged, blue dye complex that stains basophilic structures.
• Eosin is a negatively charged, pink dye that stains acidophilic (also known as eosinophilic)
structures.

Hydrophobic structures (such as those rich in fats) do not stain well with H&E.

Hematoxylin and eosin are the most widely used dyes in histology and pathology. The following
slides demonstrate the staining characteristics of these dyes alone, and more importantly, in
combination.
Hematoxylin and eosin (or H&E) are the most widely used dyes in histology. They stain many
components found in the cytoplasm, nucleus, and extracellular matrix.

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Hematoxylin is a positively charged, blue dye complex that stains
components of cells and tissues that are negatively charged.

• Phosphate groups of nucleic acids (DNA and RNA)

• Sulfate groups of some extracellular polysaccharides
(glycosaminoglycans) and proteins (mucus)

Tissue components that stain with basic dyes are referred to

as basophilic structures.
The overall blue color shows a general staining of this specimen
with hematoxylin.

At higher magnification (click this link or the above

image), structures with a range of staining
intensities become evident.

• Nuclei are intensely stained

• Cytoplasm of cells are lightly stained

Eosin
➢ is a negatively charged, pink dye that stains components of cells and tissues that are
positively charged.

• Amino groups in proteins (mostly side chains of lysine and arginine)

Tissue components that stain with acid dyes are referred to as acidophilic (or eosinophilic)
structures.
The overall pink color shows a general staining of this specimen with eosin.

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At higher magnification (click this link or the above image), the range of staining intensities of
structures is much less than with hematoxylin. Granules are seen in the cytoplasm of most cells.

• Nuclei are lightly stained

• Cytoplasm of most cells is intensely stained

Hematoxylin & Eosin

Staining with the combination of hematoxylin and eosin (H&E) reveals even more cellular structures
than either dye used alone.

At higher magnification (click this link or the above image), the nuclei and cytoplasm of individual cells
are easily identified. This reflects the complementary binding affinities of these dyes for many cellular
compotents.
These structures are described as:

• Basophilic - structures stained blue by hematoxylin

• Acidophilic (or Eosinophilic) - structures stained pink by eosin

Hydrophobic structures (such as those rich in fats) do not stain well with H&E.
Staining with the combination of hematoxylin and eosin (H&E) reveals even more cellular structures
than either dye used alone.

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At higher magnification (click this link or the above image), the nuclei and cytoplasm of individual cells
are easily identified. This reflects the complementary binding affinities of these dyes for many cellular
compotents.
These structures are described as:

• Basophilic - structures stained blue by hematoxylin

• Acidophilic (or Eosinophilic) - structures stained pink by eosin

Hydrophobic structures (such as those rich in fats) do not stain well with H&E.

Cells and Tissues

The cell is the basic structural and functional unit of all living organisms. Cells vary widely in size and
shape depending on their function. Microscopes are used to study cells because most cannot be
seen with an unaided eye.
It is not necessary to learn the names of specific cells and tissues for this chapter, but rather learn to
recognize variations in the size, shape, and staining properties of cells.
NUCLEAR MORPHOLOGY:
The staining pattern of nuclei can be either heterochromatic or euchromatic.

• Heterochromatin is a tightly packed form of DNA. Heterochromatic nuclei stain uniformly and
intensely with hematoxylin. This is indicative of a cell with low transcriptional activity.
• Euchromatin is a lightly packed form of DNA. Euchromatic nuclei show random clumps of
hematoxylin staining mostly at the periphery of the nucleus and the nucleolus. This is indicative of
cells with high transcriptional activity.

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Lymph nodes contain many lymphocytes. These small cells have very dark nuclei and a thin rim of pale cytoplasm. At
high magnification, examine individual lymphocytes surrounding this capillary.

• Lymphocytes are small cells of 7 to 10 µm diameter.

• They have heterochromatic nuclei and only a thin rim of cytoplasm.
• ∼75% of cell volume is the nucleus

Adipose tissue is composed of adipocytes. They are also known as adipose cells or fat cells. Examine the adipocytes in
the connective tissue surrounding the lymph node.

• These fat cells are 30 to 50 µm in diameter.

• They contain large lipid droplets that are unstained by either hematoxylin or eosin.
• Red blood cells have a diameter of ∼7.5 to 8 µm and can be used to estimate the size of other cells and nuclei.
• Since these fat cells are quite large, the tissue section often does pass through the part of the cell which contains the
nucleus.

• The largest cells in the human body are motor neurons in the spinal cord and mature ovum in the ovary. At higher
magnification, find motor neurons cut in cross section through its nucleus with or without processes (axons or
dendrites).

• Nuclei occur in all cells except mature red blood cells (RBC) and blood platelets. RBC are ~7.5 µm in diameter
and can be used to estimate sizes of other structures.

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• Since these cells are quite large, the tissue section frequently does not contain the nucleus. In this case, the nucleus is in
the plane of the section.
o These cells are of 80 to 120 µm in diameter.
• The nucleus is 20 to 30 µm in diameter and is euchromatic.
• They have a prominent nucleolus which is intensely stained with hematoxylin.
o The nucleolus is 10 to 15 µm in diameter.

• The liver is composed primarily of hepatocytes. These cells are closer in size to 'typical' cells than the large motor
neurons.

• At high magnification, examine hepatocytes sectioned through their nuclei that are near a blood vessel containing
RBC.

• (Note the RBC are poorly preserved.)

• Hepatocytes have an average cell diameter of ∼25 to 30 µm.

• Use the nearby red blood cells to estimate the size of the cells and nuclei.
• These cells have heterochromatic nuclei and may contain two nuclei (binucleate).
• The nuclear cytoplasmic ratio for these cells is ∼1:20 to 1:30.

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CELLS AND TISSUES:
Example tissues that exhibit a broad range of sizes, shapes, morphology, and staining characteristics of cells and nuclei.

Blood Cells
Blood Smear - blood consists of 99% red blood cells and 1% white blood cells.

• Red blood cells are biconcave discs without a nucleus and have a diameter of ∼7.5 to 8.0 µm.
o They do not stain with hematoxylin.
o The red color (eosinophilia) is due to the high concentration of the protein hemoglobin in these cells.
• White Blood Cells - have nuclei of various shapes, staining patterns and vary in size (#1, #2, #3) from ∼10 to 25 µm.

Trachea
Trachea - contains examples of basophilia (stains with hematoxylin) and acidophilia (stains with eosin).

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• Acidophilia (or eosinophilia) can be seen within cells such as red blood cells and duct cells. Eosin also stains the major
extracellular protein, the connective tissue protein collagen.
• Basophilia can be seen within and outside cells. Hematoxylin stains DNA and RNA within cells. It also stains other
negatively charged molecules. For example, the sulphated glycoproteins found in mucus and sulphated polysaccharides
found in the extracellular matrix of cartilage.
• This section contains many examples of nuclear size, shape (round, spindle, lenticular) and staining intensity
(euchromatic or heterochromatic).
Small Intestine
Small Intestine - In most places the surface epithelium is detached from the underlying supportive tissue.

• The surface eithelium is composed of polarized, column shaped cells with an oval nucleus found in the basal portion of
these cells.
o Polarized cells have a basal surface that rests on supportive connective tissue and an apical free surface that faces the
opposite side.
o The free surface always faces a lumen, such as, the interior of the gut or a duct.
• The cells in the lamina propria are found beneath the epithelial surface cells and are not polarized.
o Most cells have euchromatic nuclei.
o Many cells have eccentric nuclei (nuclei located not a the center of the cell).
• Smooth muscle and collagen.
o Smooth muscle cells are arranged in parallel bundles and have long spindle shaped euchromatic nuclei. These pink cells
are seen in cross section in the lower left and in cross section in the upper right. Note the appearance of the nuclei in
longitudinal and cross sections.
o The brighter red staining material is collagen and is extracellular. The cells interspersed among the collagen are
fibroblasts with heterochromatic nuclei.

Mitosis
Mitosis is a part of the cell cycle when replicated chromosomes are separated into two new nuclei. This
gives rise to daughter cells each having the same number of chromosomes as the parent cell.
An onion root tip is a rapidly growing part of an onion and thus many cells will be in different stages of mitosis. An onion
cell possesses 8 chromosomes (whereas human cells possess 46 chromosomes).
Examine the following cells in different stages of mitosis. (Observe the cell at the center of each view.)

• Interphase (#1, #2, and #3) - non-dividing cells (majority of

the cells)
• Prophase (#1, #2, and #3) - condensation of chromosomes
but not organized
• Metaphase (#1 and #2) - chromosomes line up in the center
of the cell
• Anaphase (#1, #2, and #3) - chromosomes are pulled to
opposite sides of the cells
• Telophase (#1 and #2) - chromosomes arrive at the cell
poles and will form two nuclei

LINKS FOR MORE READINGS:

http://histologyguide.org/slidebox/01-introduction.html
https://bio.libretexts.org/Bookshelves/Human_Biology/Book%3A_Human_Biology_(Wakim_and_Grewal)/10%3A_Introduc
tion_to_the_Human_Body/10.3%3A_Human_Cells_and_Tissues
https://open.oregonstate.education/aandp/chapter/4-1-types-of-tissues/
VIDEOS LINKS:
https://www.youtube.com/watch?v=i5tR3csCWYo
https://www.youtube.com/watch?v=15k5fajCN_w
https://www.youtube.com/watch?v=oe-Z9t0KBfU

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