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Development of Automated Infarct Size Measurement in TTC Stained Rat Isolated Hearts after Global Ischemia/Reperfusion
Author(s) -
Shidham Sushrut,
Nabbi Raha,
Camara Amadou K.S.,
Riess Matthias L.
Publication year - 2011
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.25.1_supplement.1130.2
Subject(s) - ischemia , cardiology , medicine , infarction , myocardial infarction
Infarct size (IS) remains the gold standard to estimate cardiac injury in animal models. In cumulative planimetry, several triphenyltetrazolium chloride (TTC) stained sections are scanned, and IS can be quantified manually (IS‐M). Using Image J 1.44i software and its ColorThreshold plugin we developed a macro for automated measurement (IS‐A). Results from both methods are compared prospectively. METHODS We analyzed 153 rat isolated hearts that were part of a study on genome‐dependent cardioprotection. Hearts were subjected to 30 min global ischemia and 2 hrs reperfusion after different cardioprotective treatments, cut into 2 mm transverse slices and stained in 1% TTC/0.1M KH 2 PO 4 buffer. Using Image J, slices were scanned on green background at 1200 dpi, infarcted areas measured manually, and IS‐M of the whole heart calculated by weighted average of individual slices. All images were also analyzed by the newly developed macro, and IS‐A calculated in the same manner. To determine the best color threshold T between infarcted/white and non‐infarcted/red tissue we varied T from 125 to 133. Data: mean±SD. Statistics: linear regression; *P<0.05. RESULTS Both methods correlated significantly for all chosen T. However, T129 provided the best correlation and lowest intercept: IS‐A = −2.89±1.99 + (*1.13±0.04 • IS‐M), R 2 = 0.82. The slope greater than 1 indicates a continuous increase of IS‐A over IS‐M with increasing IS. DISCUSSION The new method enables fast, operator‐independent IS measurements with a good correlation to the previous manual method. Disadvantages are lack of discrimination, e.g., between infarcted and connective tissue with a possible bias of IS‐A over IS‐M. Advantages are significant time‐ and cost‐savings and the ability to analyze more, i.e. thinner slices per heart providing a better longitudinal resolution. Supported by the Society of Cardiovascular Anesthesiologists.