FACULTY OF ENGINEERING AND LIFE SCIENCES

DEPARTMENT OF SCIENCE AND BIOTECHNOLOGY

COURSE CODE BBS 1224

COURSE NAME CELL AND TISSUE CULTURE

LECTURER FARRAH NAZUHA BINTI MANSOR

LAB TITLE PLANT TISSUE CULTURE LABORATORY-FACILITIES

AND EQUIPMENTS

SUBMISSION DATE 26 MAY 2023

STUDENT NAME STUDENT ID. SIGNATURE

1. PRATAYINI A/P 4221003641 Pratayini

PACKIANATHAN

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LAB ACTIVITY COURSE LEARNING OUTCOME MARKS

To understand the basic principles and requirements of

CLO 1 /20
animal/plant tissue cultures

TOTAL / 20
This assignments/experiments measures the student's ability for the following
outcomes:

Introduction
The design of an in-vitro propagation facilities grows out of compromises involving the space
available, the production capacity desired, the climate, the financial resources and certain design
principles related to the kind of work being carried out. The demonstrated facility would be
suitable for a small sized educational or research operation. The major factors limiting the
capacity of this design are the size of the clean room or laminar flow hood and the amount of
shelf space in the culture room. The basic operations to be performed are cleaning glassware,
preparation of culture media, sterilization of media and equipment, preparation of explants,
sterile transfer of cultured materials and growing in-vitro cultures to maturity. These operations
can be conveniently separated into two areas-a laboratory and a culture room.
Like taxonomy, cytology, plant physiology, etc., plant tissue culture is a subfield of plant
science. Instead, it is a group of experimental techniques for growing a lot of isolated cells or
tissues in sterile, controlled environments. The tissues or cells are taken from any portion of the
plant, such as the stem, root, or leaf, which is encouraged to exhibit totipotency (i.e., the genetic
capacity to make additional plants) and to produce more cells in culture. Different types of glass
vials with a media containing vitamins, phytohormones, and mineral nutrients are used to
develop cells or tissues. Therefore, a well-equipped laboratory is first necessary in order to
conduct the studies employing tissue culture procedures.
Plant physiology, cytology, taxonomy, and other fields of plant research, such as plant tissue
culture, are all part of plant science.There has been a significant increase in the number of
research labs using tissue culture techniques to look into various theoretical and practical aspects
of higher plants in recent years. These methods are employed outside of research as well, though.
Many commercial laboratories are making use of tissue culture techniques. Even a lot of
horticulture businesses are setting up modest units to multiply plants that are challenging to
propagate traditionally. A tissue culture laboratory's overall structure as well as the fundamental
techniques will be covered under various subheadings.

Objective(s)
Students should be able to:
1. Familiarize with the facilities and basic equipment needed for plant tissue culture technique
in the laboratory.

2. Understand the important rules acquired for maintaining aseptic conditions and to ensure
safety in the laboratory.
Materials & Methods
1. All students are going to visit Tissue Culture Laboratory at Department of Science and
Biotechnology, Faculty Engineering and Life Sciences.

2. Students have to identify the basic equipment/facilities and area of Plant In-Vitro Culture
Unit.

Results

1. pH meter

 An equipment called a pH metre is used to determine the acidity or alkalinity of a

solution,often known as pH. The unit of measurement used to express how acidic or
alkaline something is is called pH.

2. Weighing balance
 An instrument used to calculate an object's weight or mass is a weighing balance. It is a
necessary equipment in laboratories, commercial kitchens, and pharmacies and is offered
in a variety of sizes with various weighing capacities.

3. Distilled water
 Lead-acid batteries, automotive cooling systems, and other devices where mineral
buildup would be harmful are sterilised using distilled water. It is perfect for use in skin
care items, canning goods, and cleaning laboratory equipment because it doesn't contain
any other minerals or pollutants.
4. Media

 Culture medium provide the nutrients and minerals that cells and microorganisms need to
flourish in lab settings. However, the nutritional needs and environmental conditions of
various organisms vary. As a result, several culture mediums are developed to allow the
growth of a chosen cell or microorganism in laboratories.

5. Autoclave
 Autoclaves use high pressure and warmth to kill spores and germs. They are used to
sterilise lab equipment, sterilise media, and decontaminate specific biological waste.

6. Hot plates
 Hot plates are widely used for a variety of tasks, including heating samples and
conducting chemical reactions. The idea of a hot plate is straightforward: it is a flat
surface with heating components. They are suitable for use in oil or sand baths and do not
emit open flames.

7. Washing area
 A vital part of laboratory safety is hand washing. In fact, this 20-second method is
regarded as one of the most crucial steps utilised to stop chemicals, radioactive materials,
and microbiological agents from infecting you and the area around you.

8. Culture room
  Large open bay laboratories with tissue culture equipment are typically found in modern
research buildings. A tissue culture room is a space where infectious agents or cell
cultures from diverse animals or plants are handled in a biosafety cabinet.

Discussion
 Appendix 1
Table of Basic Equipment

Item Use/Function Brand’s @ Quantity Important

Company’s Rules/Procedure
Name

Brand’s Name: Labline Before we go into details of

Protect the working Company’s Name: the working principle, let us
1 Piece first understand one important
environment from dust
and other airborn component of a laminar flow
contaminants by cabinet: the HEPA filter! It
maintaining a constant, abbreviates as 'high-efficiency
unidirectional flow of particulate air filter'. It is
HEPA-filtered air over present within the cabinet in
the work area. order to make the
environment more sterile for
Laminar different processes.

Flow Hood

Provide a physical Brand’s Name: Esteem 1 Piece To decontaminate

method for disinfection Autoclaves
and sterilization. They
certain biological
Company’s Name:WSF waste and sterilize
work with a
Industries, Inc.
combination of steam, media, instruments
pressure and time.
Autoclaves operate at and lab ware
high temperature and
pressure in order to kill
microorganisms and
spores.

Autoclave

Brand’s Name: Ice 1 Piece Its effectively removes

It is an effective water Mountain Brand inorganic compounds such as
treatment method for Distilled Water metals (lead), nitrate, and
removing contaminants other nuisance particles such
like bacteria, heavy Company’s Name: as iron and hardness from a
Water metals, and chemicals. Dazzle Sparkling Water contaminated water supply

distiller

Chemical It’s used to weigh small 1 Piece It tells about the number of
 amounts of substances Brand’s Name: Intell Lab
accurately up to the HT 224 Analytical moles reacting and producing.
fourth place of decimal. Balance Also, it tells the number of
It is necessary for the electrons transferred from one
volumetric analysis as Company’s Name: compound to another.
the substances used to Popular Science Balancing/ balanced equation
prepare standard Apparatus Workshops also talks about the energy
solutions are weighed. Pvt Ltd (PSAW) changes that take place.

balance

To measure acidity or Brand’s Name: To analyze the exact pH value

Environmental Express 3 Pieces
alkalinity of a solution - of food grade products and
also known as pH. pH is (7) chemical products to ensure
the unit of measure that safety and quality, or they can
describes the degree of be used to evaluate
acidity or alkalinity. It Company’s Name: BR acidity/alkalinity of drugs in
is measured on a scale Biochem Life Sciences pharmaceutical and
pH meter of 0 to 14. Private Limited. biotechnology industries.

Defined as a device
with a flat surface and
an internal electric 1 Piece To perform chemical
heating element that is reactions, to heat samples, and
used for cooking or for numerous other activities.
heating food. Brand’s Name: Stuart Hot plates are conceptually
UC152 Hot Stirrer, max. simple a flat surface with
capacity 15L, Ceramic heating elements. They do not
produce open flames and are
Company’s Name: well suited for oil or sand bath
RUNNER-UP. use.
Hot Plate
with/without
stirrer

Can help ensure the accuracy

of an experiment or test.
Brand’s Name: SMC
To utilize the power of 1 Piece Environmental factors such as
Thermo Chiller Thermo
outside air and water to temperature can skew
chiller, HRSE018-A-23.
maintain the target research results if there is
temperature at a Company’s Name: even one degree of variation.
constant level. Kirloskar Pneumatic By accurately ensuring exact
Therefore, it can be Company Ltd. temperatures, laboratory
used to cool or heat. chillers fulfill the crucial role
Because it is necessary of guaranteeing that
to keep the temperature experiments or tests can be
constant at all times, it completed under the
Chiller is not limited to cooling. appropriate conditions.
 To mix, blend, or
agitate substances in a Brand’s Name: Ohaus. 1 Piece Aims to homogenize, disperse
tube or flask by shaking and mix samples at precise
them. It is mainly used Company’s Name: speeds and with precise
in the fields of Orbital shaker KS 501 control. The resulting sample
chemistry and biology. digital without should be uniform in
A shaker contains an accessories. composition and free of
oscillating board that is clumps, lumps, or other
used to place the flasks, imperfections that can affect
Shakers beakers, or test tubes. the accuracy of test results.

Volumetric flasks are Volumetric flasks are

for measuring accurate important for measuring
volumes of liquid accurate volumes of liquid
materials for laboratory substances in
experiments. They are experimentation. These flasks
favored when available are used when precise
because they are more measurements are critical, and
accurate than graduated can measure within the range
cylinders and beakers, of 1 ml to 10,000 ml.
which are other pieces
of equipment that are Beakers can be used to hold
used to measure liquids. liquid or solid samples or as a
container for reactions. They
Beakers are mainly used are also used to collect
for measuring and filtrates from filtering
transporting liquid, operations as well as liquids
while Erlenmeyer from titrations. It is used for
Flasks are typically stirring the liquid. It is useful
used for holding, as a reaction container
mixing, measuring,
heating, and boiling
chemicals, chemical
reactions, and phase
changes.

Volumetric NIL NIL

flasks and
Beakers
 Petri dish is a shallow
transparent glass or
plastic cylindrical
lidded dish, used to
contain a thin layer of
agar on which to grow
bacteria and fungi
Culture vessels provide
a contamination barrier
to protect the cultures
from the external
environment while
maintaining the proper
internal environment.
For anchorage-
dependent cells, the
vessels provide a
suitable and consistent
substrate for cell
Petri dishes attachment. NIL
and culture
vessels
Microscope is an
instrument that is used
to magnify small
objects. Some
microscopes can even
be used to observe an
object at the cellular
level, allowing
scientists to see the
shape of a cell, its
nucleus, mitochondria,
Microscope and other organelles. NIL
Petri dish is majorly used as
laboratory equipment in the
field of biology & chemistry.
The dish is used to culture
cells by providing storage
space and preventing them
from getting contaminated.
The culture vessels provide a
barrier against contamination
to protect the cultures from
the external environment
while maintaining an adequate
internal environment.
NIL
Microscope allowed to see
cells, bacteria, and many other
structures that are too small to
be seen with the unaided eye.
It gave them a direct view into
the unseen world of the
extremely tiny. You can get a
glimpse of that world in
Figure below.
NIL
In-vitro culture units (IVC)

Unit/Area Use/Function Major

Equipment@Facilities@Items
Found

Large sinks and draining

boards can be found
here. Additionally, there
is distilled water
available here, along
with a drying area. As a In general, most labware
result, it makes it easier should be washed with
to perform the initial detergent and water in the
cleaning necessary to sink immediately after use to
Washing remove any surface- prevent the buildup of
Area based bacteria. residue.

Media preparation room

Media Preparation Area
is the basis and backbone
of a microbiology
laboratory. It is a place
where all the media the
nutritional components
required for growing and
visualizing Equipment
microorganisms is •Brochure: MEDIACLAVE
prepared. 10L System

Transfer/Inoculation This chamber is An

commonly used to create inoculation
bacteria and fungi free chamber is
atmosphere. Moreover, a device or
we provide this enclosed
Inoculation Chamber in workspace
various dimension at cost used in
effective prices. microbiolog
Features: Trouble free y and other
performance. scientific
disciplines
to create a
sterile
environmen
t for the
handling
and transfer
of
microorgani
 sms.

Room

An incubation chamber
is a device used to grow
and maintain plants and
microbiological cell. The
incubator chamber is
designed to maintains
optimal temperature,
humidity, light, pressure,
vacuum and other
conditions such as the
carbon dioxide (CO2)
and oxygen content of
the atmosphere inside the
Culture/Incubation Room chamber. Plywood

Laboratory observation,
as opposed to naturalistic
observation, refers to
observing the behavior of
subjects that are in a
controlled environment.
Because of the controlled
environment variable
factors can be controlled
which therefore leads to
a limited number of Volumetric
Observation Area possible responses. flasks

A greenhouse is a
building with glass walls
and a glass roof.
Greenhouses are used to
grow plants, such as
tomatoes and tropical
flowers. A greenhouse
stays warm inside, even
during the winter. In the
daytime, sunlight shines
into the greenhouse and
warms the plants and air Greenhouse
Glasshouse/Greenhouse inside. staging.

Conclusion
Aseptic manipulation of plant material, preparation, sterilisation, and storage of nutrient media,
maintenance of cultures under controlled environmental conditions in culture rooms, data
collection and photographic facility, and an area designated for the accretion of cultures are some
of the major basic facilities used in tissue culture techniques that should be present in a standard
tissue culture laboratory.

References
 Murashige T and Skoog F (1962) A revised medium for rapid growth and bio-assays with
tobacco tissue cultures. Physiol Plant 15(3): 473-497.
 Redenbaugh, Keith, Bill Hiatt, Belinda Martineau, Matthew Kramer, Ray Sheehy, Rick
Sanders, Cathy Houck and Don Emlay (1992). Safety Assessment of Genetically
Engineered Fruits and Vegetables: A Case Study of the Flavr Savr Tomato. CRC Press.
p. 288.
 Ye, X; Al-Babili, S; Klöti, A; Zhang, J; Lucca, P; Beyer, P; Potrykus, I (2000).
"Engineering the provitamin A (beta-carotene) biosynthetic pathway into (carotenoid-
free) rice endosperm". Science 287 (5451): 303–5.doi:10.1126/science.287.5451.303
 UNISEL-FELS
Efective Date : 8th October 2018