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The investigation of BBB score of animal behavior in rat based on high-speed camera analysis

The investigation of BBB score of animal behavior in rat based on high-speed camera analysis. Presenter ： Bo - Ying Syu Adviser ： Dr . Pei- Jarn Chen Data ： 2013/6/5. Outline. Introduction Papers Review Material and Methods Future works References. Introduction ( 1 / 6 ).

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The investigation of BBB score of animal behavior in rat based on high-speed camera analysis

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  1. The investigation of BBB score of animal behavior in rat based on high-speed camera analysis • Presenter：Bo-YingSyu • Adviser： Dr. Pei-Jarn Chen • Data： 2013/6/5

  2. Outline • Introduction • Papers Review • Material and Methods • Future works • References

  3. Introduction ( 1 / 6 ) • Spinal Cord Injury • Incidence of 40 new cases per million people throughout the world each year • Mean age : 28.6 years old • Sensory and autonomic dysfunctions, the degree of which is characteristic for the severity of the SCI

  4. Introduction ( 2 / 6 ) • Experimental studies of pharmacology, neuropathology and pathophysiology commonly measure the motor function of rats. • The behavioural performance can be used as an index for neural disease process or therapy efficiency.

  5. Introduction ( 3 / 6 ) • Use Aneurysm-clip with spinal cord damage • Destruction of  parts T7、T8、T9 Thoracic Vertebrae

  6. Introduction ( 5 / 6 ) Measurement methods Locomotor tests Motor tests Sensory tests Sensory motor tests • Rope walk testing • Narrow beam test • Foot slip test • Primary open-field tests • BBB test • Automated walkway test • Footprint analysis test • Inclined plane • Limb hanging test • Limb grip strength test • Cold sensitivity-based tests • Withdrawal reflexes • Primary open-field tests • BBB test • Automated walkway test • Footprint analysis test Pictures form " An inclined plane system with microcontroller to determine limb motor function of laboratory animals" Pictures form " Methods for behavioral testing of spinal cord injured rats " Pictures form " Methods for behavioral testing of spinal cord injured rats "

  7. Introduction ( 5 / 6 ) • BBB test • Basso, Beatie and Bresnahan Test • Two or three researchers as a group and record score, then average these score • BBB Score Divided into 0-21, degree is to be used to evaluate the spinal cord injury.

  8. Introduction ( 6 / 6 ) • BBB Score • Scores from 0 to 7 ( Indicate the return of isolated movements in the hip 、 knee and anklejoints ) • Scores from 8 to 13 ( Indicate the return of paw placement and coordinated movements with the forelimbs ) • Scores from 14 to 21 ( Show the return of toe clearance during stepping, predominant paw position , trunk stability and tail position )

  9. Papers Review( 1 / 2 ) • PAW PRINT GAIT ANALYSIS IN RATS WITH SPINAL CORD INJURY MAY, 2007 • MALU TANSEY ,KEITH TANSEY ,SHUXIN LI

  10. Papers Review( 1 / 2 ) • Material and Methods • Surgical procedure and contusion: • Long-Evans rats (n=10) • Destruction of  parts : T9 (182 to 227 kdyn) • Postoperative : Baytril

  11. Papers Review( 1 / 2 ) • Material and Methods • High-speed video camera (fps=80) • On the treadmill • DigiGait analysis software • BBB score

  12. Papers Review( 1 / 2 ) • Material and Methods • Two to six weeks postoperatively • 5 cm/s every thirty seconds • Five consecutive strides

  13. Papers Review( 1 / 2 ) • Material and Methods Parameters were assessed: • Stride length • Stance width • Stance-to-swing ratio

  14. Papers Review( 1 / 2 ) • Results • Stride length increased 1.2-fold from the second to third week and then remained relatively constant at each time point. (stride/s at 25 cm/s)

  15. Papers Review( 2 / 2 ) A semi-automated software tool to study treadmill locomotion in the rat From experiment videos to statistical gait analysis May 2010 P. Gravela,∗, M. Tremblayb, H. Leblondb, S. Rossignolb, J.A. de Guisec

  16. Papers Review( 2 / 2 ) • Methods • Adult wistar rats (n = 6) • High-speed area scan camera (fps=120, • DS-41-300K0262 ,DALSA Inc ). • C arm X-ray system (100 kVp/16 mAs~ 40 kVp/0.5 mAs) • On the treadmill • 8~20 complete cycles

  17. Papers Review( 2 / 2 ) • Methods Image calibration • image interpolation 27 cm×27cm calibration grid

  18. Papers Review( 2 / 2 ) • Methods • Image contrast enhancement • stationary wavelet transform (order 5). • Using cubic smoothing spline ankle joint femoral head iliac crest the second toe MTP knee joint Hip angles knee angles heel angles MTP angles From top to bottom; the iliac crest, the femoral head, the knee joint, the ankle joint, the metatarsophalangeal joint (MTP), and the distal phalange of the second toe.

  19. Papers Review( 2 / 2 ) • Results The data were acquired manually without the GUI (1 h 30 min) semi-automatically with the GUI (12 min)

  20. Purpose • Use a High-speed camera to analyze animal behavior. • Combination of the BBB scores and gait parameters to measure the rat activity behavior.

  21. Material and Methods • High-speed cameras (fps=120, scA640-120gc, BASLER Inc) • Streampix 5 (Norpix Inc.) • Windows XP、C++ builder、Opencv1.0

  22. Material and Methods

  23. Material and Methods • Side scan(Marker) Cb色板 YCbCr

  24. Material and Methods • Side scan(Marker) Morphology A a c x B C b

  25. Material and Methods • Side scan(Centroid) Morphology (Gray) (Erode&Dilate) (Binarization)

  26. Material and Methods • Bottom scan(Centroid) (Gray) (Binarization) (Erode&Dilate)

  27. Material and Methods • Bottom scan(Limbs) (ROI)

  28. Results • Angle Analysis

  29. Future works • Improve the bottomscan. • Brain injury • Shoot the two sides. • Gait analysis and centroid combination to find correlations. • BBBscoremore correct.

  30. References • Methods for behavioral testing of spinal cord injured rats , Jirˇı´ Sˇ edy´ a,b, Lucia Urdzı´kova´ a,b, PavlaJendelova´ a,b, Eva Sykova´ a,b , Received 15 March 2007; received in revised form 9 August 2007; accepted 3 October 2007 • Automatic analysis of altered gait inarylsulphatase A-deficient mice in the open field , Toon Leroy, StijnStroobants , Jean-Marie Aerts , Rudi D’Hooge, and Daniel Berckmans , Behavior Research Methods , 2009, 41 (3), 787-794 • Functional evaluation of peripheral nerve regeneration in the rat : walking track analysis , Artur S.P. Varejfio a,* Marcel F. Meek b Antdnio J.A. Ferreira °, JoS.oA.B. Patricio d Antdnio M.S. Cabrita e , Journal of Neuroscience Methods 108 (2001) 1-9 • An inclined plane system with microcontroller to determine limb motor function of laboratory animals, Ming-Wen Chang, Ming-Shing Young, and Mao-Tsun Lin, Journal of Neuroscience Methods., 168: 186–194, 2008. • Robotic Assessment of Locomotor Recovery in Spinal Contused Rats , J.A.Nessler,D.J.Reinkensmeyer, K.Sharp,E.Kwak1,K.Minakata,R.D.DeLeon,26th Annual International Conference of the IEEE EMBS • Robot Applied Stance Loading Increases Hindlimb Muscle Mass and Stepping Kinetics in a Rat Model of Spinal Cord Injury, Jeff A.Nessler, MoustafaMoustafa-Bayoumi, Dalziel Soto, Jessica E. Duhon, Ryan Schmitt,33rd Annual International Conference of the IEEE EMBS

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