Lecture 3

By

Dr. Ahmed Metwaly

TISSUE CULTURE

Objectives: Cell Culture Equipments

■ Basic Equipments

■ Expanded Equipment

■ Additional Supplies

Cell Culture Equipments Basic Equipments

■ Cell culture hood (i.e., laminar-flow hood or biosafety cabinet)

■ Incubator (humid CO2 incubator recommended)

■ Water bath

■ Centrifuge

■ Refrigerator and freezer (–20°C)

■ Cell counter

■ Inverted microscope

■ Liquid nitrogen (N2) freezer

■ Sterilizer (autoclave)

Cell culture hood ( laminar-flow hood or biosafety cabinet)

How does laminar hood flow work?

■ laminar flow is the case occurs when a fluid or air flows in

parallel layers, with no disruption between the layers.

■ Laminar flow hoods are used to exclude contaminants by

passing a sterile air from inside to outside.

Incubator Incubator is a device used to

grow and maintain cell

cultures. The incubator

maintains optimal

temperature, humidity and

other conditions such as

the carbon dioxide (CO2)

and oxygen content of the

atmosphere inside.

■ The purpose of the incubator is to provide the appropriate

environment for cell growth.

■ The incubator should be large enough for your laboratory needs,

have forced air circulation, and should have temperature control .

■ Stainless steel incubators allow easy cleaning and provide

corrosion protection, especially if humid air is required for

incubation.

Types of Incubators Types of Incubators

There are two basic types of incubators, dry incubators and humid CO2

incubators.

■ Dry incubators are more economical, but require the cell cultures to

be incubated in sealed flasks to prevent evaporation. They do not

allow precise control of atmospheric conditions in the incubator.

■ Humid CO2 incubators are more expensive, but allow superior control

of culture conditions. They can be used to incubate cells cultured in

Petri dishes or multi-well plates, which require a controlled

atmosphere of high humidity and increased CO2 tension.

Centrifuge

A centrifuge is a piece of equipment

that puts an object in rotation

around a fixed axis (spins it in a

circle), applying a potentially strong

force (outward). The centrifuge

used for sedimentation of insoluble

substances.

Water bath freezer (–20°C)

Most cell culture reagents can be stored at –5°C to –20°C;. A

domestic freezer is a cheaper alternative to a laboratory freezer.

Cell counter or hemacytometer

■ Automated cell counters sample the culture, quantify,

classify, and describe cell populations using both

electrical and optical techniques.

■ A cell counter is essential for quantitative growth

kinetics.

Inverted microscope ■ An inverted microscope is

a microscope with its light source and condenser on the top, above the stage pointing down, while the objectives and turret are below the stage pointing up.

■ Inverted microscopes are useful for observing living cells at the bottom of a large container (e.g., a tissue culture flask) under more natural conditions than on a glass slide, as is the case with a conventional microscope.

Liquid nitrogen or cryostorage container

Liquid nitrogen

is nitrogen in a liquid state

at an extremely low

temperature.

Liquid nitrogen is

a cryogenic fluid that can

cause rapid freezing on

contact with living tissue.

Sterilizer (autoclave)

An autoclave is a pressure

chamber used to carry out

sterilization processes under

elevated temperature and

pressure .

Expanded Equipment

■ • Aspiration pump

■ • pH meter

• Aspiration pump pH meter

■ Confocal microscope

Confocal microscopy is an optical imaging technique for increasing optical

resolution and contrast of a micrograph. It enables the reconstruction of three-

dimensional structures from the obtained images.

■ Flow cytometer

Flow cytometry is a laser-based, biophysical technology employed in cell

counting, cell sorting, biomarker detection and protein engineering, by

suspending cells in a stream of fluid and passing them by an electronic

detection apparatus.

Additional Supplies

■ • Cell culture vessels (e.g., flasks, Petri dishes, roller bottles,

multi-well plates)

■ • Pipettes and pipettors

■ • Syringes and needles

■ • Waste containers

Summary: Cell Culture Equipments

■ Basic Equipments

■ Expanded Equipment

■ Additional Supplies