coupling ferrybox and automated flow cytometer: a new ... · •few phytoplankton community dataset...

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

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).

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)

(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.

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)


variability





…



Well constrained


variability





…



Well constrainedIndividual process contributions

difficult to assess

CHROME project


variability





…



Mediterranean Sea

• Only few pCO2 data (SOCAT, LDEO databases)

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

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

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

CHROME project


& composition


FerryBox


PhD ThesisYann Bozec & Pascal Morin

CHROME project


& composition


FerryBox


Melilotus Thyssen & Gérald Grégori

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

SubCtech OceanPack MK2:• Flat-Membrane-Equilibrator• LI-COR LI-840x inside• Auto-zeroing calibration• Span gas calibration

MIO /CHROME sensors

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

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

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




Total FWS Total SWS

Tota

l FLR

Tota

l FLO


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




Thanks to recent advances, we can now observe both with the CytoSense, in addition to larger phytoplankton cells.

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

Image in flow (IIF) acquisition (>20 µM)

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.

Unfortunately no pCO2 data due to technical issues.

Chl-a conc. (µg L-1)

Cryptophytes like Pico-Eukaryotes Nano-Eukaryotes

Synechococcus

Measurements every 20 minutes

Prochlorococcus

Microphytoplankton

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


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)

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

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


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)

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


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

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.

coupling ferrybox and automated flow cytometer: a new ... · •few phytoplankton community dataset...

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