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Group 1 :

Aseptic Work Area

Teh Ai Yeen 260110103004Gan Soon Chong 260110103008

Sivakanth Selvamani 260110103015

Rubananthan Ramasamy 260110103021

Jegatheeswaran Krishnan 260110103026

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Introduction : Tissue Culture

A method of biological research in which

fragments of tissue from an animal or plant

are transferred to an artificial environment in

which they can continue to survive and

function.

Facilitated via use of a liquid, semi-solid, or

solid growth medium, such as broth or agar

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Involves exposing tissue to a specific regimen

of nutrients, hormones, and light under

sterile, in vitro conditions to produce many

new clone of the original mother, over a very

short period of time

Involve 3 stages :

Initiation

Multiplication

Root Formation

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Stage 1

It concerns the establishment of plant tissue in

vitro by sterilising the material and initiating it into

culture.

Stage 2

The in vitro plant material is re-divided and placed

in a medium with plant growth regulators that

induce the proliferation of multiple shoots. Repeated many times until the number of plants

desired is reached.

Stage 3

Involves the introduction of hormones to induce

rooting and the formation of complete plantlets.

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Aseptic Work Area

Successful cell culture depends heavily on

keeping the cells free from contamination by

microorganisms such as bacterial, fungi, and

viruses.

Nonsterile supplies, media, and reagents,

airborne particles laden with microorganisms,

unclean incubators, and dirty work surfacesare all sources of biological contamination.

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Aseptic technique provide a barrier between

the microorganism in the environment and

the steril cell culture.

Cell culture hood is normally used.

Cell culture hood reduce contamination fromaerosol and airborne particles.

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Cell Culture Hood

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Air is drawn through a HEPA filter and blown

in a very smooth, laminar flowtowards theuser. The cabinet is usually made of stainless

steel with no gaps or joints where sporesmight collect.

Such hoods exist in both horizontal andvertical configurations, and there are many

different types of cabinets with a varietyof airflow patterns and acceptable uses.

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The cell culture hood should be properly set

up and be located in an area that is restricted

to cell culture that is free from drafts from

doors, windows, and other equipment, and

with no through traffic.

The work surface should be uncluttered and

contain only items required for a particular

procedure; it should not be used as a storage

area.

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Before and after use, the work surface should

be disinfected thoroughly, and the

surrounding areas and equipment should be

cleaned routinely.

For routine cleaning, wipe the work surface

with 70% ethanol before and during work,

especially after any spillage.

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You may use ultraviolet light to sterilize the air

and exposed work surfaces in the cell culturehood between uses.

Using a Bunsen burner for flaming is notnecessary nor is it recommended in a cellculture hood.

Leave the cell culture hood running at alltimes, turning it off only when they will not beused for extended periods of time.

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