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Automation of Vessel Counting. Jessica DeQuach Boris Babenko Christman Lab Belongie Lab. Death of cells. Degradation of ECM. Wall thinning. Negative left ventricular remodeling. Heart Failure. Myocardial Infarction and Heart Failure. Heart Attack.
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Automation of Vessel Counting Jessica DeQuach Boris Babenko Christman Lab Belongie Lab
Death of cells Degradation of ECM Wall thinning Negative left ventricular remodeling Heart Failure Myocardial Infarction and Heart Failure Heart Attack
Myocardial Infarction and Heart Failure RV LV 6 weeks later….
Research With Injectable Biomaterials • Want to develop an injectable biomaterial to help regenerate/salvage damaged tissue • Skeletal muscle • Heart • Injectable materials • Minimally invasive (less surgery) • Contours to shape of wound
Animal Surgeries • Surgeries performed by Pam or Rebecca • Animals anesthetized • First surgery: injury (infarct, ischemia) • Second surgery: injection, ~1 week later • Animals kept in vivarium until time point http://kentsimmons.uwinnipeg.ca/16cm05/16labman05/lb8pg12.htm
Animal Studies (weeks to months) • To get statistical significance, usually need n >= 4 per group • Need to account for animal death, and variability • Need to compare to a control group • Note* need to include different experimental groups on each day
Analysis • Various methods to determine whether the material is helping the damaged area • Function • Neovascularization • Wall thickness • Cellular infiltration MRI
Neovascularization • Formation of new blood vessels with red blood perfusion http://www.omegagenesis.com/html/Research.html
Animal Studies (weeks) • Harvest tissue, freeze in OCT • Section the sample to 10 um slices
Injection Region • Identify the injection region • Stain five evenly spaced slides through the injection region H&E
Vessel Stain • Antibody stain for vessels • Antibodies can identify specific proteins • Smooth muscle actin (red) • Stains for smooth muscle cells • Isolectin (green) • Stains for endothelial cells http://www.siumed.edu/~dking2/crr/cvguide.htm
Animal Studies (weeks) • Take images of vessel stain • Need to quantify! • The annotator should be trained to be consistent • Blinded so that no bias • Only one person can quantify a set of data
Vessel Quantification • Criteria: • Within infarct or injection region • Has a lumen • Average diameter > 10 um
Vessel Quantification Identify injection region 858_82_16
Vessel Quantification Identify injection region 854_123_70
Vessel Quantification Identify injection region 879_192_17
Vessel Quantification Draw long and short axis
Vessel Quantification Not all pictures will be that pretty 872_112_21
Vessel Quantification • Vessel long and short axis are averaged • Only count vessels > 10 um • Hundreds of images, takes hours • Only 1 person can quantify
Automation • Automation of the process would be beneficial • Need to determine “human error” • This is where you come in! • Quantify 150 images of skeletal muscle and heart. Will need MATLAB, can do this from home • Need data from several people
Computer Vision (Belongie Lab) • Combination of Artificial Intelligence, Machine Learning, Image Processing, 3D Geometry (and sometimes Cognitive Science) • Examples in Industry: • Face detection (i.e. most P&S cameras today) • Google Goggles / TinEye
Computer Vision (Belongie Lab) • Pedestrian Detection • Reading text “in the wild”
Computer Vision and Bio • Smart Vivarium(Piotr Dollar et al.) • Coral Classification (Oscar Beijbom et al.)
Training • We will first label 10 images together • Then you will label 10 image on your own • After someone double checks the answer you will be okay to quantify on your own
Commitment • Quantify 150 images of cardiac and skeletal muscle, only in the Region of Interest (ROI) • The Matlab program will automatically save your data even if it crashes • Can pace yourself, measure 10 or so a day • Opportunity to participate in Bioengineering Research, and can come to lab meetings, etc.
Contacts • Jessica DeQuach • jdequach@ucsd.edu • Boris Babenko • bbabenko@ucsd.edu • Anthony Monteforte • amontefo@ucsd.edu