Digitally quantifying cerebral hemorrhage using Photoshop and Image J.
A spectrophotometric hemoglobin assay is widely used to estimate the extent of brain hemorrhage by measuring the amount of hemoglobin in the brain. However, this method requires using the entire brain sample, leaving none for histology or other assays. Other widely used measures of gross brain hemorrhage are generally semi-quantitative and can miss subtle differences. Semi-quantitative brain hemorrhage scales may also be subject to bias. Here, we present a method to digitally quantify brain hemorrhage using Photoshop and Image J, and compared this method to the spectrophotometric hemoglobin assay. Male Sprague-Dawley rats received varying amounts of autologous blood injected into the cerebral hemispheres in order to generate different sized hematomas. 24h later, the brains were harvested, sectioned, photographed then prepared for the hemoglobin assay. From the brain section photographs, pixels containing hemorrhage were identified by Photoshop and the optical intensity was measured by Image J. Identification of hemorrhage size using optical intensities strongly correlated to the hemoglobin assay (R=0.94). We conclude that our method can accurately quantify the extent of hemorrhage. An advantage of this technique is that brain tissue can be used for additional studies.
Tang
Xian Nan
XN
Department of Neurology, University of California, San Francisco & SF VAMC, San Francisco, CA 94121, USA.
Berman
Ari Ethan
AE
Swanson
Raymond Alan
RA
Yenari
Midori Anne
MA
D000818:Animals; D001921:Brain / Q000473:pathology*; D001923:Brain Chemistry; D002543:Cerebral Hemorrhage / Q000473:pathology*; D004195:Disease Models, Animal; D006454:Hemoglobins / Q000032:analysis; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D016014:Linear Models; D051381:Rats; D017207:Rats, Sprague-Dawley; D012984:Software; D013053:Spectrophotometry
D006454:Hemoglobins
Stroke. 2009 May;40(5):1858-63
19286595
Comput Med Imaging Graph. 2009 Jun;33(4):304-11
19269786
Annu Int Conf IEEE Eng Med Biol Soc. 2008;2008:1214-7
19162884
Stroke. 1990 Nov;21(11):1589-93
2122552
Stroke. 2002 Oct;33(10):2478-84
12364741
J Neuroimaging. 1997 Oct;7(4):227-31
9344005
Neuroreport. 2001 Feb 12;12(2):309-11
11209940
Neuroscience. 2008 Jun 23;154(2):556-62
18511205
Stroke. 1997 Nov;28(11):2296-302
9368579
J Nucl Med. 2007 Nov;48(11):1822-8
17942809
J Pharmacol Exp Ther. 1993 Jan;264(1):67-73
8423552
J Cereb Blood Flow Metab. 1997 Apr;17(4):401-11
9143222
J Stroke Cerebrovasc Dis. 1998 May-Jun;7(3):179-86
17895078