FerryBox systems at HZG: Experiences and applications on ...
Coupling FerryBox and automated flow cytometer: a new ... · •Few phytoplankton community dataset...
Transcript of Coupling FerryBox and automated flow cytometer: a new ... · •Few phytoplankton community dataset...
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Coupling FerryBox and automated flow cytometer: a new approach for studying biogeochemical processes and CO2 system variability in
relation to phytoplankton community structure
Heraklion, 6 and 7 April 2016
Pierre Marrec1, Cherif Sammari2, Nagib Bhairy1, Mathilde Dugenne1, GéraldGrégori1, Sana Ben Ismail2, Soumaya Lahbib1 and Melilotus Thyssen1
1 CNRS/INSU, IRD, Mediterranean Institute of Oceanography, Aix Marseille Université, Marseille, France2 INSTM, National Institute of Marine Sciences and Technologies, Salammbô, Tunisia
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Atmospheric CO2 concentration for the past 2000 years
0 500 1000 1500 2000
CO
2 c
on
cen
trat
ion
(p
pm
)
260
280
300
320
340
360
380
400
1800 ≈ 280 ppmGlacial/Interglacial
Maximum
2014 ≈ 400 ppm
+40%
Since the industrial revolution: atmospheric CO2 has increased by ≈40%.
Highest atmospheric CO2 concentration over at least the last 800 000 years.
Essentially due to anthropogenic activities.
Increase of atmospheric GHG concentration (CO2, as well as CH4 and N2O) is “extremely likely” (95-100% probability) the main driver of climate change (IPCC, 2013).
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The world ocean plays a fundamental role in the exchange of CO2 with the atmosphere and constitutes an important sink for atmospheric CO2.
The ocean stores nearly 1/3 of anthropogenic carbon released to the atmosphere (Sabine et al., 2004).
Physical and biological processes control the ocean carbon cycle with the solubility pump and the biological pumps.
(IPCC report, 2007)
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(IPCC report, 2007)
Essential to improve our understanding of the dynamics of the CO2 system and to quantify air-sea CO2 fluxes for
global carbon cycle and climate studies.
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Seawater partial pressure of CO2 (pCO2)
variability
Thermodynamic processes:- Solubility
Non-thermodynamic processes:- Biological processes
- Dissolution/formation of CaCO3- Lateral advection
- Horizontal mixing- Upwelling
…
Seawater temperature/salinity Structural phytoplankton heterogeneityPresence of calcifying organisms
3D oceanic circulation (observations and models)
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Seawater partial pressure of CO2 (pCO2)
variability
Thermodynamic processes:- Solubility
Non-thermodynamic processes:- Biological processes
- Dissolution/formation of CaCO3- Lateral advection
- Horizontal mixing- Upwelling
…
Seawater temperature/salinity Structural phytoplankton heterogeneityPresence of calcifying organisms
3D oceanic circulation (observations and models)
Well constrained
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Seawater partial pressure of CO2 (pCO2)
variability
Thermodynamic processes:- Solubility
Non-thermodynamic processes:- Biological processes
- Dissolution/formation of CaCO3- Lateral advection
- Horizontal mixing- Upwelling
…
Seawater temperature/salinity Structural phytoplankton heterogeneityPresence of calcifying organisms
3D oceanic circulation (observations and models)
Well constrainedIndividual process contributions
difficult to assess
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CHROME project
Seawater partial pressure of CO2 (pCO2)
variability
Thermodynamic processes:- Solubility
Non-thermodynamic processes:- Biological processes
- Dissolution/formation of CaCO3- Lateral advection
- Horizontal mixing- Upwelling
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Seawater temperature/salinity Structural phytoplankton heterogeneityPresence of calcifying organisms
3D oceanic circulation (observations and models)
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Mediterranean Sea
• Only few pCO2 data (SOCAT, LDEO databases)
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Mediterranean Sea
• Only few pCO2 data (SOCAT, LDEO databases)
• Few phytoplankton community dataset resolving the submeso- & mesoscale : • space scales of 1-10 km and time scales of 1-10 days• space scales of 10-100 km and time scales of 10-100 days
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Mediterranean Sea
• Oligotrophic ecosystem
• Dominated by small phytoplankton (picoplankton):• Cyanobacteria : Prochlorococcus & Synechococcus (0,6 – 1,2 µm)• PicoEukaryotes (< 2 µm)
• Importance of picoplankton in oligotrophic areas:• major primary producers• most abundant phytoplankton organisms on earth
ProchlorococcusBiodiversity project – Austin Chiang
SynechococcusPhoto: John Waterbury,
Woods Hole Oceanographic Institute
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CHROME project
High-frequency pCO2Phytoplankton abundance
& composition
Biogeochemical and physical parameters
FerryBox
Last technological advances Scientific expertise
Unravel the role of phytoplankton community structure on biogeochemical variability and CO2 system dynamics at high spatio-
temporal resolution
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CHROME project
High-frequency pCO2Phytoplankton abundance
& composition
Biogeochemical and physical parameters
FerryBox
Last technological advances Scientific expertise
PhD ThesisYann Bozec & Pascal Morin
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CHROME project
High-frequency pCO2Phytoplankton abundance
& composition
Biogeochemical and physical parameters
FerryBox
Last technological advances Scientific expertise
Melilotus Thyssen & Gérald Grégori
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1 Marseille ↔ Tunis& 1 Genova ↔ Tunis
crossings per week.
Continuous measurements: Temperature Salinity Dissolved Oxygen Fluorescence Turbidity
Our sensors CHROME/MIO pCO2 pH Automated Flow
Cytometer (FC)
FerryBox installation: February 2016pCO2, pH and FC implementation: March 2016But the system is not entirely operational yet
C/F Carthage
INSTM FerryBox system
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SubCtech OceanPack MK2:• Flat-Membrane-Equilibrator• LI-COR LI-840x inside• Auto-zeroing calibration• Span gas calibration
MIO /CHROME sensors
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SubCtech OceanPack MK2:• Flat-Membrane-Equilibrator• LI-COR LI-840x inside• Auto-zeroing• Span gas calibration
CytoBuoy CytoSense Flow Cytometer• Single cell analysis• Automated and remotely
controlled• Image in flow acquisition• Run with embedded computer
MIO /CHROME sensors
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SubCtech OceanPack MK2:• Flat-Membrane-Equilibrator• LI-COR LI-840x inside• Auto-zeroing calibration• Span gas calibration
CytoBuoy CytoSense Flow Cytometer• Single cell analysis• Automated and remotely
controlled• Image in flow acquisition• Run with embedded computer
Satlantic SeaFET:• ion sensitive field
effect transistor (ISFET)
• Our first deployment
MIO /CHROME sensors
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Thanks to recent advances, we can now observe prochlorococcus with the CytoSense, in addition to larger phytoplankton cells.
Total FWS Total SWS
Tota
l FLR
Tota
l FLO
FWS: forward scatter, SWS: sideward scatter, FLR: red fluorescence, FLO: orange fluorescence
Synechococcus
Prochlorococcus
ProchlorococcusBiodiversity project – Austin Chiang
SynechococcusPhoto: John Waterbury,
Woods Hole Oceanographic Institute
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Total FWS Total SWS
Tota
l FLR
Tota
l FLO
FWS: forward scatter, SWS: sideward scatter, FLR: red fluorescence, FLO: orange fluorescence
It will allow us to better understand the spatio-temporal dynamics of these major photosynthetic contributors in relation to air-sea CO2 fluxes.
Synechococcus
Prochlorococcus
ProchlorococcusBiodiversity project – Austin Chiang
SynechococcusPhoto: John Waterbury,
Woods Hole Oceanographic Institute
Thanks to recent advances, we can now observe both with the CytoSense, in addition to larger phytoplankton cells.
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10 phytoplankton groups defined:ProchlorococcusSynechococcusPicoplankton & Picoplankton High FLRCryptophytes likeNanoplankton 1 & Nanoplankton High FLOMicroplankton & Microplankton High FLOUnidentified (for the moment)
FLR5 FLR30
Calibration with beads (1-10 µm) to classphytoplankton clusters according to their size, anduse of pictures for microphytoplankton (>20 µm).
Total SWS (a.u.)
Tota
l FLR
(a.
u.)
Total SWS (a.u.)
Tota
l FLR
(a.
u.)
Tota
l FLO
(a.u
.)
Total SWS (a.u.)
Example of clusteringOSCAHR campaign
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Image in flow (IIF) acquisition (>20 µM)
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Pre-FerryBox automated flow cytometer deployment: OSCAHR
Observing Submesoscale Coupling At Hight Resolution, PIs Andrea Doglioli &Gérald Grégori, 2015-10-29 to 2015-11-06 onboard the RV Tethys II.
Last generation Cytosense flow cytometer with IIF deployment with a -4H-Pocket FerryBox (SST, SSS, O2, FLUO) and a subCtech pCO2 sensor.
Similar configuration as in the CHROME project.
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Unfortunately no pCO2 data due to technical issues.
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Chl-a conc. (µg L-1)
Cryptophytes like Pico-Eukaryotes Nano-Eukaryotes
Synechococcus
Measurements every 20 minutes
Prochlorococcus
Microphytoplankton
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Day
Night
All
Total Fluorescence from the Cytometer (a.u.)
Flu
ore
scen
ce f
rom
th
e Fl
uo
rim
eter
(a.
u.)
Red fluorescence (FLR): 1 of the optical parameter recorded by the cytometer (FLO, SWS,FWS).
Strong correlation between: • The sum of the total red
fluorescence recorded for all the clusters
• And the fluorescence recorded by the research vessel fluorimeter
• Quenching during daytime
FWS: forward scatter, SWS: sideward scatter, FLR: red fluorescence, FLO: orange fluorescence
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Leg2
03/11 06:00 03/11 18:00 04/11 06:00 04/11 18:00 05/11 06:00 05/11 18:00 06/11 06:00
SSS
37,9
38,0
38,1
38,2
38,3
38,4
SST
(°C)
15
16
17
18
19
20
03/11 06:00 03/11 18:00 04/11 06:00 04/11 18:00 05/11 06:00 05/11 18:00 06/11 06:00
Diss
olved
O2 c
once
ntrati
on (µ
mol k
g-1 )
200
210
220
230
240
O 2 sa
turati
on le
vel (%
)
86
88
90
92
94
96
98
Chl-a
conc
entra
tion (
µg L-
1 )
0,1
0,2
0,3
0,4
0,5
Abun
danc
e (N
/mL)
10000
20000
30000
40000
50000
Synechococcus
Prochlorococcus
Abun
danc
e (N
/mL)
600
800
1000
1200
1400
1600
1800PicoEukaryotes
NanoEukaryotes
03/11 06:00 03/11 18:00 04/11 06:00 04/11 18:00 05/11 06:00 05/11 18:00 06/11 06:00
Abun
danc
e (N
/mL)
20
40
60
80
100
120
500
1000MicroEukaryotes
Cryptophytes
Unidentifiedflr5
Salin
ity
Dis
solv
edO
2(µ
M)
SST (°C)
Ch
l-a(µ
g/L)
O2
saturatio
n (%
)
Ab
un
dan
ce(N
mL-
1)
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Leg2
03/11 06:00 03/11 18:00 04/11 06:00 04/11 18:00 05/11 06:00 05/11 18:00 06/11 06:00
SSS
37,9
38,0
38,1
38,2
38,3
38,4
SST
(°C)
15
16
17
18
19
20
03/11 06:00 03/11 18:00 04/11 06:00 04/11 18:00 05/11 06:00 05/11 18:00 06/11 06:00
Diss
olved
O2 c
once
ntrati
on (µ
mol k
g-1 )
200
210
220
230
240
O 2 sa
turati
on le
vel (%
)
86
88
90
92
94
96
98
Chl-a
conc
entra
tion (
µg L-
1 )
0,1
0,2
0,3
0,4
0,5
Abun
danc
e (N
/mL)
10000
20000
30000
40000
50000
Synechococcus
Prochlorococcus
Abun
danc
e (N
/mL)
600
800
1000
1200
1400
1600
1800PicoEukaryotes
NanoEukaryotes
03/11 06:00 03/11 18:00 04/11 06:00 04/11 18:00 05/11 06:00 05/11 18:00 06/11 06:00
Abun
danc
e (N
/mL)
20
40
60
80
100
120
500
1000MicroEukaryotes
Cryptophytes
Unidentifiedflr5
Salin
ity
Dis
solv
edO
2(µ
M)
SST (°C)
Ch
l-a(µ
g/L)
O2
saturatio
n (%
)
Ab
un
dan
ce(N
mL-
1)
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Leg2
03/11 06:00 03/11 18:00 04/11 06:00 04/11 18:00 05/11 06:00 05/11 18:00 06/11 06:00
SSS
37,9
38,0
38,1
38,2
38,3
38,4
SST
(°C)
15
16
17
18
19
20
03/11 06:00 03/11 18:00 04/11 06:00 04/11 18:00 05/11 06:00 05/11 18:00 06/11 06:00
Diss
olved
O2 c
once
ntrati
on (µ
mol k
g-1 )
200
210
220
230
240
O 2 sa
turati
on le
vel (%
)
86
88
90
92
94
96
98
Chl-a
conc
entra
tion (
µg L-
1 )
0,1
0,2
0,3
0,4
0,5
Abun
danc
e (N
/mL)
10000
20000
30000
40000
50000
Synechococcus
Prochlorococcus
Abun
danc
e (N
/mL)
600
800
1000
1200
1400
1600
1800PicoEukaryotes
NanoEukaryotes
03/11 06:00 03/11 18:00 04/11 06:00 04/11 18:00 05/11 06:00 05/11 18:00 06/11 06:00
Abun
danc
e (N
/mL)
20
40
60
80
100
120
500
1000MicroEukaryotes
Cryptophytes
Unidentifiedflr5
Salin
ity
Dis
solv
edO
2(µ
M)
SST (°C)
Ch
l-a(µ
g/L)
O2
saturatio
n (%
)
Ab
un
dan
ce(N
mL-
1)
Phytoplankton functional and structural heterogeneity is now described at submesoscale and can be related to
physical structures
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First Results: 1 Marseille-Tunis transect 24-25/03/2016
Projected latitude (°N)
37 38 39 40 41 42 43Ch
l-a co
nc. (
µg L
-1)
0,0
0,5
1,0
1,5
2,0
xCO 2
(ppm
)
340
350
360
370
380
390
SST
(°C)
13,5
14,0
14,5
15,0
15,5
16,0
Salin
ity
37,0
37,5
38,0
38,5
Salin
ity
xCO
2(p
pm
)
5,0e+3
1,0e+4
1,5e+4
2,0e+4
2,5e+4
2e+3
4e+3
6e+3
8e+3
1e+4
PicoEukaryote
NanoEukaryotes
2e+4
4e+4
6e+4
8e+4
1e+5
Prochlococcus
Synechococcus
Projected latitude (°N)
37 38 39 40 41 42 43
0
50
100
150
3e+4
4e+4
5e+4
6e+4
7e+4
MicroEukaryotes
Cryptophytes Like
Phytoplankton Group Abundance
Flu
o (
a.u
.)SS
T (°
C)
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First Results: 1 Marseille-Tunis transect 24-25/03/2016
Projected latitude (°N)
37 38 39 40 41 42 43Ch
l-a co
nc. (
µg L
-1)
0,0
0,5
1,0
1,5
2,0
xCO 2
(ppm
)
340
350
360
370
380
390
SST
(°C)
13,5
14,0
14,5
15,0
15,5
16,0
Salin
ity
37,0
37,5
38,0
38,5
Salin
ity
xCO
2(p
pm
)
5,0e+3
1,0e+4
1,5e+4
2,0e+4
2,5e+4
2e+3
4e+3
6e+3
8e+3
1e+4
PicoEukaryote
NanoEukaryotes
2e+4
4e+4
6e+4
8e+4
1e+5
Prochlococcus
Synechococcus
Projected latitude (°N)
37 38 39 40 41 42 43
0
50
100
150
3e+4
4e+4
5e+4
6e+4
7e+4
MicroEukaryotes
Cryptophytes Like
Phytoplankton Group Abundance
Flu
o (
a.u
.)SS
T (°
C)
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0%
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
36,89 37,62 38,25 39,06 39,88 40,69 41,49 42,28 43,15
Prochlorococcus Synechococcus PicoEukaryotes
NanoEukaryotes Cryptophytes Like MicroEukaryotes
Projected latitude (°N)
37 38 39 40 41 42 43Ch
l-a co
nc. (
µg L
-1)
0,0
0,5
1,0
1,5
2,0
xCO 2
(ppm
)
340
350
360
370
380
390
SST
(°C)
13,5
14,0
14,5
15,0
15,5
16,0
Salin
ity
37,0
37,5
38,0
38,5
Co
ntr
ibu
tio
n t
o t
ota
l re
dfl
uo
resc
en
ce
Salin
ity
xCO
2(p
pm
)
Flu
o (
a.u
.)SS
T (°
C)
First Results: 1 Marseille-Tunis transect 24-25/03/2016
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Nice
Reinstallation of the sensors as soon and as long aspossible.
High-resolution phytoplankton structure heterogeneity.
Essential information of phytoplankton contribution topCO2 variability and biogeochemical processes.
Expectation of a huge dataset to get new insights aboutsurface Mediterranean ecosystems.
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Nice