Equipment
Chapter 5
Laminar Flow Hood
Horizontal Laminar Flow Hood
Suitable for preparing medium, sterile reagents and culturing nonprimate cells
Suitable for dissecting nonprimate cells
Class II or Class III Cabinet
Hazardous materials – any primate, human cell lines, virus-producing cultures, radiosiotopes or carcinogenic or toxic drugs
Follow local and national biohazard regulations before installation
Measurements of a laminar hood
Working surface are of 1200 mm (4 ft) wide x 600mm (2 ft) deep
Quiet Cleaning both inside working area and below
work surface to avoid spillage Comfortable to sit Flexibility with front screen (never remove)
Pipette Cylinders
Made up of polypropylene
One per workstation – freely distributed in the laboratory
Allow 2 hours stand of pipettes in disinfectants
Aspiration Pump
Suction pressure – remove medium or other reagents from culture flask
Effluent – discharged into container (hypochlorite added)
Switch pump before use
Vacuum pump
Same principle
Waste collected in container (emptied)
Switch pump before use
Avoid using contaminated vacuum lines
Service Carts
Place all items on service carts
Carry soiled materials and restock hoods
Inverted Microscope
Look at cultures Good quality optics Long-working distance
between condenser and objectives
Provision of CCD camera and monitor
Fluorescence microscopes
Centrifuges
Increase concentration of cells
Wash off reagents
High speed – damage to cells and agglutination
Large-scale suspension cultures – refrigerated centrifuge – 4 x 1L or 6 x 1L
Sterile Liquid Handling – Pipetting and Dispensing
Removal of fluids Rubber bulb is cheap and
simple use Pipetting aids – weight
and feel of instrument Pipettors/Micropipette Pipettes and tips
Dispensing mechanisms
Large-volume dispensing (> 100 ml)
100 ml pipette or graduated bottles or bags or > 500 mls (peristaltic pumps – different flows – also for Repetitive dispensing)
Single fluid transfers (10-10,000 L)
Multipoint Pipettor
Fill many vials at one time (based on the number of tip holding capacity)
Plate Filler
Automatic filling device
Load multiple plates
Sterile or nonsterile conditions
Cell Counter
Hemocytometer – counts number of cells
Cell counter – automatic counting of cells in solution
Incubator
Culture cells Safety thermostat Resistant to corrosion –
clean Shelving is perforated Routine maintenance of
culture is important Uniform density exp-
place in metal trays
Humid Co2 Incubator Humidifying tray – controls CO2 tension CO2 monitoring device – draws air from
incubator into sample chamber, determines concentration of CO2 and injects pure CO2
Gold wires or Infrared detectors reset zero of CO2 detector
Cannot culture flasks – use hot rooms gassed with CO2 supply
Frequent cleaning is essential – dismantle shelves
Water-jacketed with circulating fan and Dry-walled with no circulating fan
Differences
Water-jacketed Water jacket distributes
heat evenly around cabinet
Hold temperature longer in power failure
Air circulation by fan – keeps CO2 level and temperature uniform
Dry-walled Incubators No water jacket
Cannot hold temperature longer in power failure
No fan
Less fungal contamination on walls
Magnetic Stirrer
Used for suspension cultures
Speed = controlled @ 50 rpm
Capable of stirring 10 L fluid
Preparation and Sterilization
Wash up – Soaking baths or sinks
- Pipette washer – Connected to Deionized water (final rinse) and regular tap cold water
- Pipette drier – Dry pipettes (rack or drying oven)
Water Purifier
Water Purifier
Rinsing glassware – Deionized or reverse osmosis water
Powdered media and diluting concentrations – Ultra pure water
Reverse osmosis – charcoal filtration- deionization-micropore distillation
Purified water should not be stored. Must be recycled continuously to avoid growth of microorganisms
Autoclaves
Other equipment
Balances, pH meter, Hot plate magnetic stirrer, automatic dispenser, conductivity meter (ionic concentration), refrigerator and freezer, Computer and networks, PCR cycler, syringes and needles, sterilization filters, paper towels and swabs, disinfectants (chlorine based disinfectants) etc
Glassware Washing Machine
Water for washing should be heated to 80 degree C
Last step – rinsing with deionized water - 50-60 degree C
- Avoid cracking of dishes- Recycle water should be discarded- Drier is recommended
Specialized Equipment
Microinjection facilities – can inject directly into cell – example nuclear transplantation or dye injection
Colony Counter – Cell counting Centrifugal elutriator – Special centrifuge to
separate cells of different sizes Flow Cytometer – Counts cell populations based
on light scatter, absorbance and fluorescence
This project is funded by a grant awarded under the President’s Community Based Job Training Grant as implemented by the U.S. Department of Labor’s Employment and Training Administration (CB-15-162-06-60). NCC is an equal opportunity employer and does not discriminate on the following basis:
against any individual in the United States, on the basis of race, color, religion, sex, national origin, age disability, political affiliation or belief; and
against any beneficiary of programs financially assisted under Title I of the Workforce Investment Act of 1998 (WIA), on the basis of the beneficiary’s citizenship/status as a lawfully admitted immigrant authorized to work in the United States, or his or her participation in any WIA Title I-financially assisted program or activity.
Disclaimer
This workforce solution was funded by a grant awarded under the President’s Community-Based Job Training Grants as implemented by the U.S. Department of Labor’s Employment and Training Administration. The solution was created by the grantee and does not necessarily reflect the official position of the U.S. Department of Labor. The Department of Labor makes no guarantees, warranties, or assurances of any kind, express or implied, with respect to such information, including any information on linked sites and including, but not limited to, accuracy of the information or its completeness, timeliness, usefulness, adequacy, continued availability, or ownership. This solution is copyrighted by the institution that created it. Internal use by an organization and/or personal use by an individual for non-commercial purposes is permissible. All other uses require the prior authorization of the copyright owner.
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