BIO203 Laboratory 1 Plant Cells and Tissues

Objectives
Be able to identify and describe:
1. Parenchyma, collenchyma and sclerenchyma cells and their distinguishing characteristics
2. Xylem tissue: primary vs. secondary xylem, vascular bundles, protoxylem, metaxylem, tracheids,
vessel elements
3. Phloem tissue: primary vs. secondary phloem, companion cells, sieve tubes, sieve plates, fibres

Overview
In lecture you learned that plants are composed of three main organ groups: stems, roots and leaves.
These organs are made up of tissues working to accomplish a goal, or function, e.g. photosynthesis,
transport, or storage. In this lab you will look at the different cell types that make up these tissues and
learn to identify them based on their structural characteristics. The different tissues we will focus our
attention on can be classified into two broad categories: simple tissues and complex tissues. Simple
tissues are made up of one of three different cell types: parenchyma, collenchyma or sclerenchyma.
Complex tissues are composed of multiple cell types working together to accomplish one function.
Xylem and phloem tissue are important complex tissues that primarily function in the transport of water,
ions, and sugars throughout plants. Today you will learn to identify the three simple tissues and also
observe different structures of xylem and phloem tissues.

There are several ways of observing cell types and characterizing their structure in plant organs. Today
you will be hand-sectioning the tissue and staining it with Toluidine Blue O (TBO). TBO is a polychromatic
dye, which means that different components of the cell wall stain different colours. Walls containing
pectic substances but not lignin stain a pink-purple colour, whereas walls containing lignin stain
various shades of blue or blue-green.

PART 1: Cell types in simple tissues

1.1 Parenchyma cells

Parenchyma cells are the most abundant cell type
and are found in all major organs of plants. They can
carry out numerous functions, including
photosynthesis, food and water storage, secretion and
short distance transport. They assume different
shapes depending on their position in organs and their
function. In leaves, parenchyma cells usually contain
chloroplasts, and form a type of tissue called
chlorenchyma, whose main function is to perform
photosynthesis. Parenchyma cells are living and
pluripotent, which means that they have the ability to
differentiate into other cell types.
Parenchyma cells have a nucleus, cytoplasm, and
one or more large vacuoles. The cells have a thin,
usually unlignified, primary cell wall that stains Figure 1. Parenchyma cells have a nucleus (N), a cytoplasm (C),
pink-purple with TBO because of the abundance of and one or more large vacuoles (V) and a thin unlignified primary
pectic substances. Intracellular spaces are usually cell wall (CW) that stains pink-purple with TBO. Intracellular
spaces (IS) are also usually visible. Image from Peterson et al.
visible in the tissue (see Figure 1). 2008.

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BIO203 Laboratory 1 Plant Cells and Tissues

1.2 Collenchyma cells

Collenchyma cells are differentiated
parenchyma cells. They are usually located
at the periphery of aerial organs like stems
and petioles, right beneath the epidermis.
They provide flexible support for growing and
mature organs. They contain living
protoplasm and may remain alive for a long
time.

Collenchyma cells are characterized by an

unevenly thickened cell wall, which is very
conspicuous under the microscope due to its
higher refractive index. The pattern in which
the cell wall is deposited around the cell is
the basis for classifying collenchyma as
angular, lacunar or lamellar. Angular
collenchyma cells have wall thickening in their
Figure 2. Collenchyma cells may be classified as angular (AN), lacunar
corners and stain pink-purple with TBO. (LC) or lamellar (LM) depending on the shape of the cell wall.
Lacunar collenchyma cells are similar except Intercellular spaces (IS) may or may not be present between the cells.
that intercellular spaces are still present. Collenchyma cells usually stain pink-purple with TBO. Image from
Lamellar collenchyma cells have thickenings Peterson et al. 2008.
on the inner and outer tangential cell walls
(see Figure 2).

1.3 Sclerenchyma cells

Sclerenchyma cells are found in most plant
organs and have a thick, tough, secondary
wall that is impregnated with lignin. Most
sclerenchyma cells are dead at maturity and
their main function is to provide structural
support to plant organs.
Two forms of sclerenchyma cells occur:
sclereids and fibres. Sclereids can assume a
variety of shapes but are usually as long as
they are wide. They may be grouped within
other tissues or occur in specific zones or
regions. Pit canals are usually visible in the
thick, secondary cell wall. Fibres are usually
elongated, narrow cells. Each sclerenchyma
cell has a tiny cavity, called the lumen, in the
center of the cell. Because of the high lignin
content of the cell walls, sclerenchyma cells Figure 3. Sclerenchyma cells may occur as sclereids or fibres. Sclereids
appear blue or blue-green when stained with have a primary (PW) and a thick lignified secondary cell wall (SW) that stains
blue or blue-green when stained with TBO. Pits (P) are usually present in the
TBO (see Figure 3). secondary wall. Fibres are elongated, narrow cells. Image from Peterson et
al. 2008.

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BIO203 Laboratory 1 Plant Cells and Tissues

Exercise 1: Identifying Simple Tissues

Materials:
1 compound light microscope
1 pair dissecting forceps
Digital microscope eyepiece camera
Paint brush
Distilled water with a dropper
Toluidine Blue O with a dropper
1 Petri dish (2 halves)
Kimwipes
Razor blade
Microscope slides
Glass coverslips
Section of celery (Apium graveolens var. dulce) stem
Slice of pear (Pyrus communis) fruit

Methods:
1. Label a clean glass slide with: species name, organ (e.g. stem, fruit), and c.s. for cross section or
l.s. for longitudinal section (for example: P. communis, fruit, c.s.).
2. Cut a thin cross-section of a celery stem, using a straight edge razor blade. You may find it easier to
cut either against a cutting board or by holding the celery in one hand and carefully slicing across the
top. The aim is to get a very thin, very even slice. The slice should be approximately half a square
centimeter in size (small enough to fit under a cover slide comfortably). Cut several sections at a time
as it may take a few tries to get an even slice. Sections will vary in thickness but, there will be usable
ones among the "thick" sections. Transfer sections to water in a petri dish (so they do not dry out)
using a brush or forceps.
3. Select the thinnest section then use a pair of dissecting forceps (or a wet paint brush) to gently
transfer the tissue section to the center of a glass slide for staining. Try to pick it up by a corner or the
very edge; do not squeeze the tissue as you will end up crushing the cells.
4. Place 1-2 drops of TBO on the section so that it covers the whole slice. You can gently tilt the slide so
that it is immersing the entire slice.
5. After 60 seconds, remove the TBO gently with a Kimwipe. Do not put the Kimwipe on your tissue
slice, just put the corner next to it and allow it to absorb.
6. Place 1-2 drops of distilled water on the slice (this will rinse off any remaining TBO) and draw it off
with a Kimwipe again.
7. Place 1 drop of distilled water on the slice and put a cover slip on top. When placing a cover slip, put
one edge against the glass and gently lower in one direction so that the tissue adheres to the glass
surface and minimizes the risk of trapping any air bubbles.
8. Repeat steps 1-7 with the slice of pear. (This may be easier to cut against the cutting board.)
N.B. When examining your slides under the microscope, experiment with different
magnifications (4X, 10X, 40X), as well as the fine-tune knob in order to look at the 3D structure
of cells particularly for angular collenchyma. Be careful and do not turn the microscope to
the oil lens (100X) as it may hit the glass slide and damage either the slide or the lens.

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BIO203 Laboratory 1 Plant Cells and Tissues

I. Examine celery (Apium graveolens var. dulce) stem sections and locate and label parenchyma
cells, as well as angular collenchyma cells arranged in bundles. Using the microscope
camera available (1 per group of 4 students), take pictures of your specimen and make notes
using the caption tool in the software. In your lab notebook, also make notes to accompany the
pictures you take for future studying purposes. Make note of the:
a. cell wall (CW) and intracellular spaces (IS) surrounding parenchyma cells (PA), as
well as angular collenchyma cells (AN), using 40X magnification. Note the
differences in thickness of cell walls and lignification.

II. Examine pear (Pyris communis) fruit sections and locate and label parenchyma cells, as well
as sclereids arranged in clusters. Using the microscope camera available (1 per group of 4
students), take pictures of your specimen and make notes using the caption tool in the software.
In your lab notebook, also make notes to accompany the pictures you take for future studying
purposes. Make note of the:
a. cell wall (CW) and intracellular spaces (IS) surrounding parenchyma cells (PA), as
well as pits (P) observable in the thickened walls of sclerenchyma cells (SC), using
40X magnification.

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BIO203 Laboratory 1 Plant Cells and Tissues

PART 2: Complex tissues

2.1 Xylem
Xylem tissue is an important component of the vascular system of plants as it is the main conducting
tissue through which water and minerals are transported from the roots to all plant organs. During
development, all vascular plants develop primary xylem from procambium in the apical meristems
located in root and shoot tips. Primary xylem is composed of protoxylem, which forms earlier during
plant development, and metaxylem, which forms later after the elongation/expansion of the plant organ is
complete. Plants that undergo secondary growth form secondary xylem that is produced by the
vascular cambium. Secondary xylem cells eventually become dead and empty, losing their conducting
functions and serving only to support the plant.

Both primary and secondary xylem tissues

consist of a combination of parenchyma cells,
fibres, vessels, tracheids and ray cells.
Vessels are long tubes composed of
individual cells, called vessel members,
which have thick secondary cell walls and are
opened at each end. A perforation plate may
be found between the end walls of pairs of
vessel members, and can show various
patterns, most commonly bark-like strips.
Tracheids are cells with a thick cell wall and
are tapered at the end. The tips of tracheids
are usually overlapping and a pair of pits is
usually present wherever two tracheids are in
Figure 4. Tracheids and vessel members are the main types of cells that
contact with one another, allowing for water to form the xylem. Water is conducted from one vessel member to another
pass from one cell to another. Vessels and through perforation plates, and circulates from one tracheid to another via
tracheids are together referred to as pits. Image from Peterson et al. 2008.
tracheary elements (see Figure 4).

While most conduction in the xylem is vertical, ray cells allow for some lateral conduction. Ray cells are
horizontal rows of parenchyma cells that develop from vascular cambium and radiate out from the center
of stems and roots like the spokes of a wheel.

Tracheary elements exhibit different cell wall patterns

depending on the time at which the elements were formed.
Tracheary elements that first develop in the protoxylem have
either helical (spiral) or annular secondary wall patterns.
This allows these elements to stretch and elongate.
Treachery elements in the metaxylem have scalariform,
reticulate or pitted secondary cell wall patterns and usually
have ceased elongation (see Figure 5).

Figure 5. Tracheary elements that first develop in the protoxylem have

either helical (spiral) or annular secondary wall patterns while those in
the metaxylem have scalariform, reticulate or pitted secondary cell wall
patterns. Image from Peterson et al. 2008.

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BIO203 Laboratory 1 Plant Cells and Tissues

2.2 Phloem
Phloem tissue conducts and distributes products of photosynthesis (photosynthate: predominantly
sugars) throughout the plant. All vascular plants develop primary phloem that differentiates from the
procambium. Plants that undergo secondary growth develop secondary phloem from the vascular
cambium. Both types of phloem consist of a number of cell types.

Phloem is composed primarily of two types of cells lacking secondary walls, sieve elements and
companion cells. Sieve elements are the photosynthate conducting cells. They may be either sieve cells
or sieve tube members depending on the plant group. Sieve tubes are formed when two or more sieve
elements align end to end, separated by sieve plates allowing for photosynthate movement through
pores. Sieve tube members are associated with narrower, more tapered parenchyma cells called
companion cells. Fibres are also present in phloem; their secondary cell walls function primarily to
augment structural support of the tissue (see Figure 6).

Figure 6. Phloem tissue is composed of sieve elements (SE) aligning end to end to form sieve tubes. Sieve elements are
separated by porous sieve plates (SS). Companion cells (CC) are usually associated with sieve tube members. Fibres (F) can
also be present in phloem, providing support to the tissue. These structures are usually visible in both cross-section (left panel)
and longitudinal section (right panel). When examining longitudinal sections, slime plugs (SP), solid materials originating from
collapsed protoplasts (P), might be visible in the vicinity of the sieve plates. Image from Peterson et al. 2008.

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BIO203 Laboratory 1 Plant Cells and Tissues

Exercise 2: Identifying Xylem and Phloem Tissue

Materials:
1 compound light microscope
1 pair dissecting forceps
Digital microscope eyepiece camera
Distilled water with a dropper
Toluidine Blue O with a dropper
1 Petri dish (2 halves)
Kimwipes
Razor blade
Microscope slides
Glass coverslips
Section of coleus (Plectranthus scutellarioides) stem
Section of tomato (Solanum lycopersicum) or begonia (Begonia obliqua) stem (only one will be
available)

Methods:
1. Prepare and stain a cross-section and a longitudinal section of a coleus stem. Place these on two
separate slides (you may have to make multiple longitudinal sections). Follow the general method as
described in Exercise 1, remembering to always label your slides accordingly. N.B., while in Exercise 1
you only did cross section mounts, here in Exercise 2 you must do both cross section mounts and
longitudinal section mounts of the coleus stem and the tomato or begonia stems.

2. Prepare and stain a cross-section and a longitudinal section of a tomato or begonia stem. Again, place
these on two separate slides (you may have to make multiple longitudinal sections).

I. Examine a stem of coleus (Glycine max). Using the microscope camera available (1 per group
of 4 students), take pictures of your specimen and make notes using the caption tool in the
software. In your lab notebook, also make notes to accompany the pictures you take for future
studying purposes. Make note of the:
a. the cross-section of a vascular bundle using 10X magnification, as well as
regions where the xylem and phloem are located. Try to identify and label regions
of protoxylem or metaxylem.
b. the longitudinal section of a vascular bundle using 40X magnification, locating
any tracheary elements, and identify with labels the type of secondary wall
patterns that you observe. Label whether your vascular bundle is composed of
protoxylem or metaxylem.
c. In both slides, try to locate and label sieve tubes (ST), sieve plates (SS),
companion cells (CC), and/or fibres (F).

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BIO203 Laboratory 1 Plant Cells and Tissues

II. Examine a cross-section of a stem of tomato or begonia (Cucumis sativa). Using the
microscope camera available (1 per group of 4 students), take pictures of your specimen and
make notes using the caption tool in the software. In your lab notebook, also make notes to
accompany the pictures you take for future studying purposes. Make note of the:
a. the cross-section of a vascular bundle using 10X magnification, as well as
regions where the xylem and phloem are located. Try to identify and label regions
of protoxylem or metaxylem.
b. the longitudinal section of a vascular bundle using 40X magnification, locating
any tracheary elements, and identify with labels the type of secondary wall
patterns that you observe. Label whether your vascular bundle is composed of
protoxylem or metaxylem.
c. In both slides, try to locate and label sieve tubes (ST), sieve plates (SS),
companion cells (CC), and/or fibres (F).

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