W orking Group on Acoustic and Egg Surveys for Sardine and Anchovy in I CES Areas VI I I and I X, Nantes, 2 4 -2 8 / 1 1 / 2 0 0 8 Automating batch fecundity measurements Automating batch fecundity measurements using digital image analysis systems. using digital image analysis systems. A case study in the Iberian sardine. A case study in the Iberian sardine. Ganias K. 1 , Vavalidis T. , Vavalidis T. 1 , Ganias K. , Nunes C. 2 , Stratoudakis Y. , Stratoudakis Y. 2 Nunes C. 1 School of Biology, Aristotle University of Thessaloniki, 54 124 Thessaloniki, Greece, 2 Instituto Nacional de Recursos Biologicos, INRB/ IPIMAR, Avenida de Brasilia s/ n, 1449-006, Lisboa, Portugal
Processing of ovarian subsamples for Processing of ovarian subsamples for batch fecundity measurements batch fecundity measurements 2 nd subsample : microphotograph of whole mount 1 st subsample : Direct counting of oocytes under the stereoscope (Classic method)
On screen counting of hydrated oocytes using the On screen counting of hydrated oocytes using the ‘Cell counter’ ‘Cell counter’ plugin/ImageJ plugin/ImageJ (reference method reference method)
Automatic procedure for counting hydrated Automatic procedure for counting hydrated oocytes through ImageJ( oocytes through ImageJ(ImageJ ImageJ method method) Opening of image file
Automatic procedure for counting hydrated Automatic procedure for counting hydrated oocytes through ImageJ( oocytes through ImageJ(ImageJ ImageJ method method) Adjusting brightness/ contrast
Automatic procedure for counting hydrated Automatic procedure for counting hydrated oocytes through ImageJ( oocytes through ImageJ(ImageJ ImageJ method method) Converting to 8-bit image type
Automatic procedure for counting hydrated Automatic procedure for counting hydrated oocytes through ImageJ( oocytes through ImageJ(ImageJ ImageJ method method) Thresholding
Automatic procedure for counting hydrated Automatic procedure for counting hydrated oocytes through ImageJ( oocytes through ImageJ(ImageJ ImageJ method method) Segmentation of particles
Automatic procedure for counting hydrated Automatic procedure for counting hydrated oocytes through ImageJ( oocytes through ImageJ(ImageJ ImageJ method method) Counting of particles at the size range of hydrated oocytes
Automatic procedure for counting hydrated Automatic procedure for counting hydrated oocytes through ImageJ( oocytes through ImageJ(ImageJ ImageJ method method) Making the eggs look more like eggs…
Comparison between the results of the different Comparison between the results of the different counting methods counting methods 2800 3100 2700 IJM (ooc/g) CM (ooc/g) 2400 2300 2000 1900 1600 1500 1200 1100 1200 1600 2000 2400 2800 1200 1600 2000 2400 2800 RM (ooc/g) RM (ooc/g) (X 10000) (X 10000) 6 6 F-IJM (oocytes) 5 F-CM (oocytes) 5 4 4 3 3 2 2 1 1 0 0 0 1 2 3 4 5 6 0 1 2 3 4 5 6 (X 10000) (X 10000) F-RM (oocytes) F-RM (oocytes)
And now… And now… some conclusions some conclusions This method may… •increase accuracy in batch fecundity fecundity measurements • contribute in saving time and work-load (oocyte and work-load (oocyte counting 5 to 8 times counting 5 to 8 times faster compared to the compared to the classic method) method) •provide large archives of digital digital microphotographs for future analysis future analysis
Ευχαριστούμε πολύ ! …many thanks in Greek
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