901303 Cytometer Technologies for Autonomous Platforms : Comparisons between cytometers, comparison to other methods
This page last changed on Sep 09, 2014 by oreilly.
Comparison between existing oceanographic cytometers
The below characteristic values are found in various sources, including peer-reviewed publications and manufacturers marketing literature (e.g. specification sheets)
| Imaging Flow CytoBot (Heidi Sosik, McLane Research) |
SeaLabel (Jarred Swalwell, UW) |
CytoSub (CytoBuoy BV) |
FlowCam (Fluid Imaging) |
LISST 100X |
LISST Holo (Sequoia Scientific) |
LOPC | Submersible holographic microscope (Resolution Optics) |
|
|---|---|---|---|---|---|---|---|---|
| type | imaging, pulse |
pulse |
imaging (optional), pulse |
imaging (includes chl, phyco fluor channels) |
nephelometer | holographic imaging |
pulse |
holographic imaging |
| cell size range |
1-100 micron |
0.5-20 micron (SeaFLow) 2-20 micron (SeaLabel) |
0.4-700 micron ("pico" option) |
10 - 600 micron |
25 micron-2.5 mm |
100 micron - 3.5 mm |
1 micron-2 mm (depends on particle opacity, optical setup) |
|
| cell concentration range |
10^3-10^9 cells/liter |
< 10^3 particles/liter |
||||||
| instrument dimensions | 26 x 102 cm |
17 x 23 x 28 cm |
37 x 75 cm |
46 x 38 x 66 cm | 76.7 x 13.3 cm | 30 x 10 cm |
||
| instrument mass (air/water) |
32 kg |
65 kg |
18 kg (without housing?) |
9.5/3.6 kg |
11.4/6.0 kg |
1.81/1.39 kg |
||
| power |
35 W |
35 W |
sampling: 60 W (full options) idle: 20 W sleep: 2 W |
40-60 W continuous |
4.5 W sampling? |
< 20 W |
~5 W (not incl. computer) |
|
| max depth |
40 meter |
shipboard | 200 meter |
200 meter |
300 meter |
660, 3400, 6000 meter |
100, 6000 meter |
|
| Fluor/scatter excitation |
635 nm laser |
457 nm laser |
various lasers |
532 nm laser |
670 nm laser |
670 nm laser |
N.A. | |
| flow rate thru measurement volume |
15 ml/hr |
900 ml/hr f(vcore) ~ 90 ml/hr f(OPP) ~ .9 ml/hr |
0.25-64.8 ml/hr |
With 4X Objective and 300µm Flow Cell: Up to 180 ml/hour (50 mm^3/s) WIth 10X Objective and 80µm Flow Cell: Up to 45 ml/hour (12.5 mm^3/s) |
||||
| Detectors | 2 PMTs: CHL (680), SSC (635) |
5 PMTs: 2 position sensitive detect CHL1 CHL2 Phyco-erythrin |
FSC Photodiode SSC PMT CHL PMT (up to 7 add. channels) |
CCD | ||||
| max throughput | ? pulse/sec 167 images/min |
24,000 pulse/sec |
5000 pulse/sec 1500 images/min |
10,000 images/min |
2 Hz (analysis post-sample) |
16 fps 2048x2048 50 fps 512x512 |
||
| sheath fluid? |
yes | no |
yes | no | no | no | no | |
| unattended duration |
6 months "routinely" |
? | ? | ? | ||||
| realtime data processing |
yes | not implemented | yes | requires human operator, windows PC |
||||
| cost | ||||||||
| Notes on measurement method |
Olson and Sosik claim scatter/fluorescence is usually not sufficient for genus/species-level identification of nano- and micro-plankton. |
Size histogram for particles 100-1500 micron; shape info for particles > 1.5 mm |
||||||
| References | specification sheet Olson et al (2003) Olson and Sosik (2007) |
CytoAUV 2013 proposal |
Thyssen et al (2008) specification sheet |
Web page specification sheet |
specification sheet |
Herman et al (2008) Web page specifications, integration guide |
specification sheet |
|
| Event rate, time, energy to measure 1000 cells (100 cells/ml)* |
0.417 pulse/sec 2400 sec 84 kJoule |
2.5 pulse/sec (detected) 40000 sec (OPP) 1400 kJoule |
f=64.8 ml/hr: 1.8 pulse/sec 555 sec 33 kJoule |
5 - 20 particles/sec 200 - 800 sec 12 - 48 kJoule (60 W) |
||||
| Event rate, time, energy to measure 1000 cells (10^5 cells/ml)* |
417 pulse/sec 2.4 sec 84 Joule |
2500 pulse/sec (detected) 40 sec (OPP) 1400 Joule |
f=64.8 ml/hr: 1800 pulse/sec 0.55 sec 33.3 Joule |
5000-20000 pulse/sec 0.2 - 0.8 sec 12 - 48 Joule |
||||
| Notes | Has been integrated with Dorado |
Has been integrated with Dorado AUV |
Hobson investigating for possible AUV integration |
* Time to measure N cells, where p is cell number density (i.e. concentration) and f is flow rate through measurement volume:
If N = 1000, coefficient of variation = 3.1% and SNR = 32.
(See Bellingham's notes on cytometer performance.)
Comparison to other autonomous methods
| ESP | Fluorometry |
LISST 100x |
Cytometer |
|
|---|---|---|---|---|
| Advantages | Species ID |
Fast sampling; bulk pigments |
Species ID; straightforward data processing of pulse signals (image analysis more complex) |
|
| Drawbacks | Limited by onboard reagents; long sampling time |
Doesn't discriminate species |
Doesn't discriminate cells from other particles; complex data processing; must assume particle transmittance, shape; calibration problematic |
Complex optics, hydraulics; precise optical alignment. Indirect cluster-based species ID for non-imaging cytometers |
| Sample rate |
~hourly? |
~1 Hz |
KHz |
|
| Processing | simple |
complex | Based on statistical clustering of light scatter/fluorescence correlation |
|
| Species ID? |
yes |
no | no | yes |
| Samples per deployment |
10's | |||
Cytometer performance
- Linear range (signal : number of photons)
- Detector efficiency (Qr) - determined by laser power, optical design, PMT sensitivity... photoelectrons detected per equivalent fluorochrome molecule
- Optical background (Br)
- Particle size range
- Signal processing speed (pulses/sec, images/sec)
FlowCAM_Spec_Sheet_200ppi-1.pdf (application/x-octect-stream)
LISST-HOLO.pdf (application/force-download)
McLane-IFCB-Datasheet.pdf (application/force-download)
J. Plankton Res.-2008-Thyssen-333-43.pdf (application/force-download)
CytoAUV_proposal_2013-v3.pdf (application/force-download)
CytosenseSpecifications.pdf (application/x-pdf)
Submersible_Spec_Sheet_200ppi-3.pdf (application/x-pdf)
BD-characterization-performance.pdf (application/x-pdf)
FlowCytobot-Olson_et_al_DSR2003_8932.pdf (application/x-pdf)
equation.jpg (image/jpeg)
IM 230-Flow Cytometer Performance for AUV Operations.docx (application/vnd.openxmlformats-officedocument.wordprocessingml.document)
