Automating batch fecundity measurements Automating batch fecundity - - PowerPoint PPT Presentation

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. Ganias K.1, Vavalidis T. , Vavalidis T.1, , Nunes C. Nunes C.2, Stratoudakis Y. , Stratoudakis Y.2

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

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

1st subsample : Direct counting of oocytes under the stereoscope (Classic method)

2nd subsample : microphotograph

• f whole mount

Processing of ovarian subsamples for Processing of ovarian subsamples for batch fecundity measurements batch fecundity measurements

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

• ocytes through ImageJ(
• ocytes through ImageJ(ImageJ

ImageJ method method)

Opening of image file

Adjusting brightness/ contrast

Automatic procedure for counting hydrated Automatic procedure for counting hydrated

• ocytes through ImageJ(
• ocytes through ImageJ(ImageJ

ImageJ method method)

Converting to 8-bit image type

Automatic procedure for counting hydrated Automatic procedure for counting hydrated

• ocytes through ImageJ(
• ocytes through ImageJ(ImageJ

ImageJ method method)

Thresholding

Automatic procedure for counting hydrated Automatic procedure for counting hydrated

• ocytes through ImageJ(
• ocytes through ImageJ(ImageJ

ImageJ method method)

Segmentation

• f particles

Automatic procedure for counting hydrated Automatic procedure for counting hydrated

• ocytes through ImageJ(
• ocytes 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

• ocytes through ImageJ(
• ocytes through ImageJ(ImageJ

ImageJ method method)

Automatic procedure for counting hydrated Automatic procedure for counting hydrated

• ocytes through ImageJ(
• ocytes through ImageJ(ImageJ

ImageJ method method)

Making the eggs look more like eggs…

RM (ooc/g) IJM (ooc/g)

1200 1600 2000 2400 2800 1200 1600 2000 2400 2800

RM (ooc/g) CM (ooc/g)

1200 1600 2000 2400 2800 1100 1500 1900 2300 2700 3100

F-RM (oocytes) F-IJM (oocytes)

1 2 3 4 5 6 (X 10000) 1 2 3 4 5 6 (X 10000)

F-RM (oocytes) F-CM (oocytes)

1 2 3 4 5 6 (X 10000) 1 2 3 4 5 6 (X 10000)

Comparison between the results of the different Comparison between the results of the different counting methods counting methods

• 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

This method may…

And now… And now… some conclusions some conclusions

Ευχαριστούμε πολύ!

…many thanks in Greek