Counting and sizing microplastic fribres, the accurate and easy way.
Transcript of Counting and sizing microplastic fribres, the accurate and easy way.
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Counting and sizing microplastic fibres, the accurate and easy way
Kunnen, T. H.Gerber, G.
Coote, M. W.Moodley, G. K.
Robertson-Andersson, D. V.
University of KwaZulu-Natal, School of Life Science
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Introduction
• Microplastics defined as being < 5 mm (Hidalgo–Ruz et al., 2012)
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Primary microplastics Are produced for a specific purpose (eg. shower gels, cleansers)
Secondary microplastics From environmental degradation (wave action, UV exposure etc) of larger plastics
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• Global demand for plastic production is not decreasing
• Last estimated at over 311 million tonnes per year (PlasticsEurope, 2015)
• Most common source of secondary microplastics is from your own household
Introduction contd...
Polyester Lastex
Spandex
Acetate
Rayon
Nylon
Acrylic
Kevlar
Orlon
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Quick detour...filters
250 – 350 µm 600 µm
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Quick detour...filters
400 µm
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Quick detour...filters
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Introduction contd...
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550–681 part/kg!
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Introduction contd...
“ Experiments sampling wastewater from domestic washing machines demonstrated that a single garment can produce > 1900 fibres per wash.”
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For cleaner lakes and archipelago
“… waste water treatment plant ± 500 000 connected persons is estimated to receive up to 16.9 ton microplastic fibres per year…”
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Materials and Methods
3 Weeks later = 0.1276 g fibres 10 – 50/100 µm long
Ultra–violet (UV) fluorescent polyethylene–terephthalate (PET) textile (395 nm)
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White LightUV Light
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Materials and Methods contd...
• Epifluorescent microscope
• Low magnification (20 – 40 x)
• 10 – 12 fields of view taken
• Images analysed with Image Pro Plus (IPP)
• Manual counting and sizing
• Automated counting and sizing
• Length and Width data generation
• Time saving
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Width = 29.5 µmLength = 350 µm
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Width = 24.3 µmLength = 390 µm
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Width = 20.6 µmLength = 868 µm
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Fibre 1:Width = 17.5 µmLength = 754 µm
Fibre 2:Width = 20 µmLength = 534 µm
Fibre 3:Width = 21.7 µmLength = 330 µm
Fibre 4:Width = 20 µmLength = 988 µm
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Automated image analysis by Binary Segmentation and Histogram Selection
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Materials and Methods contd...
• 5 volunteers
• 50 images of microfibres
• Count Manually
• Length, width and time recorded
• Automatically using IPP
• Recorded time taken
This is 39 lines of the 270 (14.4%)
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Figure 1: Average microfibre counts per filter analyzed
Manual Automated82
82.5
83
83.5
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84.5
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85.5
Methodology
Mic
rofib
res.
mus
sel-1
p = 0.9378
Results
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Manual Automated320
330
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Methodology
Mic
rofib
re Le
ngth
(µm
)
p = 0.5478
Figure 2: Average mirofibre lengths (µm) measured utilizing manual and automated methodologies
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Manual Automated0
10
20
30
40
50
60
70
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90
Methodology
Mic
rofib
re W
idth
(µm
)
p = 0.0079
Figure 3: Average microfibre widths measured utilizing manual and automated methodologies
13.7 µm
113 µm
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AutoWidth = 50.2189 µm
ManualWidth = 29.5 µm
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AutoWidth = 110.345 µm
ManualWidth = 24.3 µm
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AutoWidth = 283.866 µm
ManualWidth = 20.6 µm
Width = Area / Length
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AutoWidth = 50.2189 µm
ManualWidth = 29.5 µm
Width equationWidth = 26.57 µm
AutoWidth = 110.345 µm
ManualWidth = 24.3 µm
Width equationWidth = 24.46 µm
AutoWidth = 283.866 µm
ManualWidth = 20.6 µm
Width equationWidth = 25.98 µm
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Manual Auto Calculated0
5
10
15
20
25
Methodology
Mic
rofib
re W
idth
(µm
)
Figure 4: Average microfibre widths measured utilizing manual and automated methodologies with the equation for new widths
p = 0.0696
23.23 ± 1.45 µm21.63 ± 0.74 µm
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Manual Automated0
200
400
600
800
1000
1200
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Methodology
Tim
e (s
)1434.52 ± 411.97 sec
60.20 ± 7.76 s
23.90 ± 6.86 mins=
Figure 5: Average time (seconds) taken to count and measure microfibres on one filter
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Fibre 1:Width = 17.5 µmLength = 754 µm
Fibre 2:Width = 20 µmLength = 534 µm
Fibre 3:Width = 21.7 µmLength = 330 µm
Fibre 4:Width = 20 µmLength = 988 µm
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ManualWidth = 19.8 µmLength = 2606 µm
Width equationWidth = 32.30 µmLength = 2402.740 µm
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Conclusion
• Using the designed macro within IPP at segmentation of 27:255
• No difference in number of fibres counted
• No difference in measured lengths
• No difference in calculated widths
• Massive time saving when doing automated analysis
M A
Length
M A
Width
M A
Time
Number of fibres
M A
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Current and Future Applications• Automated counting sizing and analysis of fluorescent microplastic fibres is ongoing research in our lab
• Mussels (Perna perna)• Sea Urchins (Tripneustes gratilla and Stomopneustes variolaris)• Mullet• Successfully been applied to 6 projects on microplastic research
• Future work to look at
• Broaden the scope and range of analysis to “naturally occurring” environmental microplastics, i.e. non–fluorescent, brown, white etc
• Work out a watershed spit that will enable the differentiation of close contact fibres
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Thank you
Any questions?
AcknowledgementsThank you to the MACE lab volunteers and to the NRF for funding this project. Thanks also go to Theo van Zyl.
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References
• Hidalgo–Ruz, V., Gutow, L., Thompson, R.C. & Thiel, M. 2012. Microplastics in the Marine Environment: A Review
of the Methods Used for Identification and Quantification. Environmental Science and Technology. 46. 3060– 3075.
• PlasticsEurope 2015. Plastics – the Facts 2015: An analysis of European plastics production, demand and waste data. Konigin Astridlaan 59, 1780 Wemmel, Belgium.
Full list of internet references available upon request