Laboratory facilities 1. Media preparation, sterilization, cleaning and storage of supplies 2. Aseptic manipulation of plant material 3. Growth of the cultures under controlled environmental conditions 4. Examination and evaluation of the cultures 5.Assembling and filling records The ideal organization will allow a separate room for each of the following functions. 1. Media preparation 3. Incubation of cultures 4. General laboratory operations 2. Aseptic procedures

Facilities should be arranged as a production line 1. The area involved with washing and storage of glassware should lead to the facilities for oven sterilization and media preparation. 2 . Materials should then move from autoclave sterilization to the aseptic transfer facility. 3. After the aseptic operations, the cultures are transferred to incubators or controlled-environment chambers. 4. The cultures should be in close proximity to the laboratory containing microscopes and facilities for evaluation of the results. 5. Discarded and contaminated cultures are transferred back to the washing area.

Arrangement of the Aseptic procedures It should not be located in the vicinity of ? scientists from other research groups airborne microorganisms Located in an area ? free from current of air with minimum of traffic If Don’t have separate sterile room then ? Laminar flow cabinet bacteriological glove box

DISHWASHING 1. AUTOCLAVE 2. DETERGENT 3. DISTILLED WATER 4. OVEN DRIED

MEDIA PREPARATION Balance sensitive to milligram quantities for weighing hormones and vitamins. Less sensitive scale may be used for weighing agar and carbohydrates.

The media reagents should be shelved near the balance for convenience.

A refrigerator and a freezer for storing stock solutions and chemicals.

A combination hot plate and magnetic stirrer for dissolving inorganic reagents.

Either a pH meter or pH indicator paper for adjusting the final pH of the medium.

Relatively large quantities of single and double- distilled water.

Sterilization equipment is an integral part of media preparation. A commercial electric stove is the most economical type of oven sterilization.

Wet – heat sterilization involves either an autoclave or a pressure cooker.

Some hormones and vitamins are sterilized by ultra filtration at room temperature.

It is also useful to have a microwave oven to dissolve agar and to melt media.

After sterilization of the culture vessels by dry heat and autoclaving the medium, the culture tubes or Petri dishes are filled in the transfer chamber.

LAMINAR FLOW CABINET

INCUBATION OF THE CULTURES 1. Cultures are grown under carefully regulated environmental conditions i.e. temperature, light and humidity. 2. This is accomplished with an incubator, plant growth chamber, or controlled environment room. 3. If cell suspensions are cultured, some type of shaker or aeration equipment will be necessary. 4. Several engineering aspects should be considered in designing a culture room. 5. Optimal environmental conditions will vary depending on the species and the purpose of the experiment.

ENGINEERING ASPECTS 1. Safety and convenience of the electrical system. 2. Air flow for uniform temperature regulation 3. Arrangement of the shelving 4. Elimination of airborne contaminants.

OPTIMAL ENVIRONMENTAL CONDITIONS OPTIMAL CONDITIONS Temperature variations Light intensity Light quality Photoperiod

ADDITIONAL NEEDS Hand lens, Dissection microscope Compound microscope Chemical hood Bench-top centrifuge Fire extinguisher, first aid kit. ADDITIONAL NEEDS

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