1. HPSS: High Pressure Serial Sampler
High Pressure Bottles functioning
Tamburini et al. AEM (2003)
High Pressure Serial Sampler (HPSS)
Bianchi et al. DSR 1999
Tamburini et al. AME 2003
 To measure deep-sea prokaryotic activities in deep-sea waters maintaining in situ conditions (pressure, temperature) until 6,000 m depth

2. HPSU: High Pressure Serial Unit (High pressure bottle on its frame)
High Pressure Sampler Unit (HPSU)
Filling valve (close)
High Pressure Sampler Unit (HPSU)
A HPSU is composed of:
A high-pressure bottle (HPB) fitted on a polypropylene frame fittable on a Sea-Bird Carousel.
A pressure sensor to monitor the hydrostatic pressure inside the HPB
An exhaust tank
A pressure accumulator to compensate the deformation of materials under high hydrostatic pressure
Presently, we are closely collaborating with Top Industry company (France) to commercialize the whole system and continue new developments.
Check-valve
High-pressure bottle
Pressure sensor
Sea-Bird® carousel
Exhaust tank

3. High Pressure Bottle (HPB)
On board
the ship
Carousel Sea-Bird
Filling valve
(close) PC
Check valve
Deck Unit
Pressure sensor
Tracer
Floating piston
HPBs sterilized are ready to take
samples.
1) Tracer can be added on the top
of the floating piston.
2) Or HPB can be used to take seawater samples maintaining the high hydrostatic pressure all the time until transfer in equipressure towards other HPBs
Distilled water (DW)
Stainless wall
PEEK coating
Exhaust tank

4. Filling and retrieval of HPBs
Filling valve
(open)
Carousel Sea-Bird
On board
the ship
Check valve PC
Decompressed
High Pressure
Deck Unit
Pressure sensor
Pressure sensor
Nozzle (0.01")
Sampling maintaining
in situ pressure condition
Until now, each sample was taken one maintained under
in situ pressure condition, the
other decompressed to estimate the effect of decompression.
The check valve permits to maintain high pressure within the HPB

5. Tranfer in pressure equilibrium
Transfer from one HPB to another one is possible using a piloted pressure generator (PPG) maintaining all the time the high hydrostatic pressure within both high pressure bottles (unitil 60 Mpa).
It is therefore possible to decline that towards other HPBs with the possibility to choice the volume transferred and take sub-samples for atmospheric analyses.
Piloted Pressure Generator

6. MIO HP-Lab container in Marseille

7. MIO HP-Lab container electrical needs
220 V
50-60 Hz
63 A
220 V
50-60 Hz
63 A
220 V
50-60 Hz
63 A (ripple current)

8. MIO HP-Lab container on board the RRV Discovery (June 2015)

9. the RRV Discovery (June 2015)
Inside of the MIO HP-Lab container on board
the RRV Discovery (June 2015)
Thermostated chamber to incubate high-pressure bottles and piloted pressure generator (60 MPa)
Bench devoted to use radiolabeled compounds (3H-, 14C).
Water baths to incubate high-pressure bottles

10. the RRV Discovery (June 2015)
Inside of the MIO HP-Lab container on board
the RRV Discovery (June 2015)
High Pressure Sampler Units (HPSUs) within water baths after recovering

11. MIO HP-Lab container
Fully equipped, the MIO HP-Lab container is composed of:
25 High-Pressure Bottles (HPBs, to recover deep-sea samples, to incubate samples, to transfer samples, to cultivate without any lost of pressure, until 40 or 60 MPa)
2 High Pressure Vessels (HPVs – 100 MPa / 200 mL for cultivation),
2 HPVs – 60 MPa/5000 mL (for cultivation)
10 HPVs – 40 MPa/ mL
2 piloted pressure generators (to regulate hydrostatic pressure until 40 and 60 MPa)
4 water baths equipped with HPB rotating system (if needed)
2 thermo – unichiller (to control temperature within the water baths, 1-20 °C)
1 thermoregulated incubator (Memmert IP750, 0 – 70°C)

12. Some references…
Tamburini C., Boutrif M.*, Garel M., Colwell R.R., and Deming J.W. (2013) Prokaryotic responses to hydrostatic pressure in the ocean - a review. Environmental Microbiology 15 (5):
Boutrif M, Garel M, Cottrell MT, Tamburini C (2011) Assimilation of marine extracellular polymeric substances by deep-sea prokaryotes in the NW Mediterranean Sea. Environmental Microbiology Reports, 3:
Tamburini C., Goutx M., Guigue C., Garel M., Lefèvre D., Charrière B., Sempéré R., Pepa S., Peterson M. L., Wakeham S., Lee C. (2009b). Effects of hydrostatic pressure on microbial alteration of sinking fecal pellets. Deep-Sea Research II 56 (18):
Tamburini C., Garcin J. & Bianchi A. (2003). Role of deep-sea bacteria in organic matter mineralization and adaptation to hydrostatic pressure conditions in the NW Mediterranean Sea. Aquatic Microbial Ecology, 32 (3):
Tamburini C., Garcin J., Ragot M., Bianchi A. (2002). Biopolymer hydrolysis and bacterial production under ambient hydrostatic pressure through a 2000 m water column in the NW Mediterranean. Deep-Sea Research II, 49 (11):
Bianchi A, Garcin J, Tholosan O (1999) A high-pressure serial sampler to measure microbial activity in the deep sea. Deep Res I 46:2129–2142
Contact : Christian Tamburini
Mediterranean Institute of Oceanography, Aix Marseille Université / CNRS / IRD, Marseille, France