Setup of Plant Tissue Culture Laboratory

• avoid locating adjacent to laboratories that handle microorganisms or insects or facilities that store seeds or other plant materials

location considerations

• avoid having plotted plants

• refrain from field or greenhouse work

• shower and change clothes

how to ensure clean cultures in lab

a. media preparation / culture evaluation / record-keeping

b. aseptic transfer area

c. environmentally controlled culture area

different work areas in a plant culture laboratory

• air vents

• high foot traffic

contamination may come from

1. Bench

2. Gas outlet

3. Hot plate and magnetic stirrer

4. Analytical and top-loading balances

5. pH meter

6. Refrigerator, freezer

7. Water purification and storage system

8. Dish-washing area

9. Storage facilities—glassware, chemicals

10. Autoclave (pressure cooker will work for small media volume)

11. Low bench with inverted light and dissecting microscopes

12. Fume hood

13. Desk and file cabinets

14. Desktop centrifuge, spectrophotometer, microwave (transformation studies and protoplast isolation)

equipment in Media preparation/culture evaluation/record-keeping area

ph meter, balances, sink, reagents, and stock solutions

what are the components of laboratory chemical bench

laboratory chemical bench

where may culture media be prepared

at least 30 mins

Culture tubes containing spent medium should be autoclaved at

low bench, table, file cabinet

essential for culture evaluation and record-keeping

1. Laminar airflow transfer hood and comfortable chair

2. Dissecting microscope

3. Gas outlet

4. Vacuum lines

5. Forceps, spatulas, scalpel, and disposable blades

equipment in Aseptic transfer area

• positive-pressure airflow

• good ventilation

separate room for transfer hood should have

• fume hood that has been thoroughly scrubbed down

• carboard boxes lined with aluminum foil or plastic containers

• HEPA (high efficiency particulate air filter) attached to plastic storage box

alternatives for laminar airflow hood

1. Shelves with lighting on a timer and controlled temperature

2. Incubators—with controlled temperature and light

3. Orbital shakers

equipment in Environmentally controlled culture area

increases contamination

High humidity in the culture room should be avoided because it

25C-27C under a 16:8-h light:dark photoperiod

Most cultures can be incubated in a temperature range