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

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

Introduction ( 1 / 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.

Introduction ( 2 / 6 )

Use Aneurysm-clip with spinal cord damage Destruction of  parts T7、 T8、 T9

Thoracic Vertebrae

Introduction ( 3 / 6 )

Measurement methods

Locomotor tests Sensory tests Sensory motor testsMotor tests

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

Rope walk testing Narrow beam test Foot slip test

Primary open-field tests BBB test Automated walkway test Footprint analysis test

Pictures form " Methods for behavioral testing of spinal cord injured rats "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 "

Introduction ( 5 / 6 )

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.

Introduction ( 6 / 6 )

BBB Score

Scores from 0 to 7( Indicate the return of isolated movements in the hip 、

knee and ankle joints )

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 )

PAW PRINT GAIT ANALYSIS IN RATS WITH SPINAL CORD INJURY

MAY, 2007

MALU TANSEY ,KEITH TANSEY ,SHUXIN LI

Papers Review ( 1 / 2 )

Papers Review ( 1 / 2 )

Surgical procedure and contusion: Long-Evans rats (n=10) Destruction of  parts : T9 (182 to 227

kdyn) Postoperative : Baytril

Material and Methods

High-speed video camera (fps=80) On the treadmill DigiGait analysis software BBB score

Papers Review ( 1 / 2 )

Material and Methods

Two to six weeks postoperatively 5 cm/s every thirty seconds Five consecutive strides

Papers Review ( 1 / 2 )

Material and Methods

Papers Review ( 1 / 2 )

Parameters were assessed: Stride length Stance width Stance-to-swing ratio

Material and Methods

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)

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

Papers Review ( 2 / 2 )

MethodsAdult 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 treadmill8~20 complete cycles

Papers Review ( 2 / 2 )

Methods

27 cm×27cm calibration grid

Image calibration

image interpolation

Papers Review ( 2 / 2 )

Methods

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.

Image contrast enhancement stationary wavelet transform (order 5). Using cubic smoothing spline

iliac crestfemoral headknee jointankle jointMTPthe second toe

Hip angles knee angles

heel angles

MTP angles

Papers Review ( 2 / 2 )

ResultsThe data were acquired manually without the GUI (1 h 30 min)

semi-automatically with the GUI (12 min)

Use a High-speed camera to analyze animal behavior.

Combination of the BBB scores and gait parameters to measure the rat activity behavior.

Purpose

High-speed cameras (fps=120, scA640-120gc, BASLER Inc)

Streampix 5 (Norpix Inc.) Windows XP、 C++ builder、 Opencv1.0

Material and Methods

Material and Methods

Side scan (Marker)

Material and Methods

YCbCr

Cb色板

Morphology

A

B C

a

b

c

x

Side scan (Marker)

Material and Methods

Side scan (Centroid)Morphology

(Gray)(Binarization)

(Erode&Dilate)

Material and Methods

Bottom scan(Centroid)

Material and Methods

(Gray)

(Binarization)(Erode&Dilate)

Bottom scan (Limbs)

(ROI)

bgrf IIII 11.059.07.0

Material and Methods

Results•Angle Analysis

Future works

Improve the bottom scan. Brain injury Shoot the two sides. Gait analysis and centroid combination to

find correlations. BBB score more correct.

References Methods for behavioral testing of spinal cord injured rats , Jirˇı´ Sˇ edy´ a,b, Lucia Urdzı´kova´ a,b, Pavla

Jendelova´ 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, Stijn Stroobants , 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.o A.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, Moustafa Moustafa-Bayoumi, Dalziel Soto, Jessica E. Duhon, Ryan Schmitt, 33rd Annual International Conference of the IEEE EMBS