Laboratory Methods

Microbiology

Lab Documentation

ODP cores have shown that bacterial life exists as deep as 1 km below the seafloor. The microbiology laboratory aboard JOIDES Resolution enables microbiologists to investigate this "deep biosphere." The laboratory contains an anaerobic chamber with rock splitting and other sampling equipment for taking uncontaminated samples from the cores, a laminar flow hood for preparing uncontaminated samples for study, and facilities for incubating samples. There will also be an epifluorescence microscope available for sample examination. Pressure vessels and -80°C freezers allow storage of samples for future study under controlled conditions onshore. The JOIDES Resolution also has facilities for conducting tracer experiments, both to quantify the extent of downhole contamination of the cores (by drilling fluid pumped down from the sea surface) and to measure the growth rates of bacteria recovered from deep beneath the sea floor.

Major Items of Microbiology Lab Equipment
Boekel Slide Moat Incubator, Fisher P/N 05-450-31
Fisher ISOTEMP Brand Low Temperature Incubator (2)
Fisher ISOTEMP Water Bath, Model 220, P/N 15-462-20 (2)
FORMA Model 926 Upright Freezer -86C
Gas Analyzer, Oxygen/Hydrogen, Model 10
Gauge Model 68, SS, P/N 54-68D-200
Hewlett Packard G 1 223A - Electron Capture Detector on a HP 5890 GC
Homogenization Flask, P/N 33994-666
Homogenizer shaft assembly for macro open blade
Homogenizer, Cyclone IQ2 P/N 33994-537
Hot Plate, Stirring, P/N 11-497-61
Hydraulic Rock Trimmer - P/N 13H4000
Laminar Flow Hood, LABCONCO 37500-01
LC Pump Model 301 Macro Stainless Steel Head (to meter tracer in drill water)
Luminometer, TD-20/20 without injector system
MARVEL Lab Refrigerator Model # 6CRF
Nalgene 16 mm Test Tube Rack, P/N 14-809-24
Pipette, Eppendorf , 0.5-10ul, P/N 05-402-86
Pipette, Eppendorf, 100-1000ul, P/N 05-402-90
Pipette, Eppendorf, 10-100 ul, P/N 05-402-88
Pipette, Eppendorf, 500-5000ul, P/N 05-402-91
Piston Seal Kit, Macro Head, P/N 120681 (to use with LC pump)
Regulator for He - Dual Stage, P/N 3104C-CGA580
Regulator, Ultra-Purity for ECD, P/N 51-19VOC-G CGA 350
Single stage Regulator (Model 23) SS, P/N 51-23S-100
Star printer system and cable, P/N 7000-9 10
Sterilizer, P/N 14-462-5
Type "A" Glove Box, Medium Size, Unheated, with auto-airlock
Vacuum Pumps, Fisher P/N 01-092-29
Vortex Mixer, Thermolyne Max Mix 11, P/N 12-814-5
Welch vacuum pump, Model 8915, S/N KK031526
Wrist Action Shaker, P/N 14-260, Model# 75

Major Items of Microbiology Lab Equipment Added on Leg 189T
Yamato Autoclave
Fisher Water Bath
Fisher Oven
Barnstead Infinity Nanopure Ultrapure Water System
Replacement Glove Bag with Gloves on Opposite Sides
Shimazu TOC-5000 with Solid Phase Analyzer
Trace Analytical Hydrogen Reduction Gas Analyzer (being evaluated)
Gas Manifold for Gassing Anaerobic Culture Tubes (WHOI)
High Pressure Reactors, Gauges, Connectors and Pump, High Pressure Equipment Co.

Microscope to be installed soon
Zeiss Axioplan with Epifluorescence Capabilities
Zeiss Axiocam, Digital Color Camera System
Zeiss MC80DX Photographic Camera

Glove Bag (Anaerobic Chamber)
There is a Coy anaerobic chamber in the microbiology lab. This is a two person, medium size model with gloves on opposite sides of the polymer glove bag. The glove bag is 5' L X 40" H X 32" D. There is an automatic air lock for taking samples in and out of the glove bag chamber. The door to the lock is 9"W X 12" H. The air lock is automated and a single push of a button automatically cycles the air lock through a programmable sequence of applying a vacuum and flushing with nitrogen, carbon dioxide and hydrogen. This results in consistent vacuum and pressure levels. There is a catalytic box with heater to control oxygen and humidity. There is a flame device for sterilizing loops. Plumbed into the glove box are both nitrogen and an anaerobic gas mixture of 5% hydrogen, 5% carbon dioxide and the balance nitrogen.

Photomicrograph of volcanic glass (right) and clay (left) treated with Hoescht dye, which turns blue in the presence of DNA. The sample came from 284 mbsf. The long dimension of the image = 0.6 mm.

Resources

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