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F38. Automated image processing system for shape recognition of single red blood cells based on out-of-focus images
Vol. 32, Nos. 2-3
Free Communications
237
F37. INVOLVEMENT OF SYMPATHETIC VASOCONSTRICTION IN L-NAME INDUCED H Y P E R T E N S I O N IN C O N S C I O U S RATS N. IIDA Department of Physiology, School of Medicine, University of Hiroshima, Minarni-ku, Hiroshima 734, Japan The aims of the study are: (1) to d e t e r m i n e w h e t h e r sympathetic vasoconstriction is involved in L-NAME (inhibitor of NO synthase) induced hypertension in conscious rats, and (2) to understand whether or not the abnormal tone caused in peripheral vascular beds is of spinal origin. For these purposes, we studied the effects of ganglion blockade hexamethonium (C6) and L-arginine on L-NAME induced hypertension in intact and spinal rats. Spinal transection was performed at Thoracic 1 in ether-anesthetized rats with an electromagnetic flow probe placed around the superior mesenteric artery, renal artery or terminal aorta, and a catheter in the jugular vein for drug application. Peripheral resistance was calculated by dividing arterial pressure by peripheral blood flow. Three or 4 days after operation, regional blood flow and arterial pressure were measured in the conscious, resting state. L-NAME injection (3-5 m g / k g iv) resulted in a significant increase in mean arterial pressure and peripheral vascular resistances in both intact and spinal rats. Ten minutes after L-NAME injection, the infusion of C6 (25 m g / k g iv) reduced arterial pressure and peripheral vascular resistances significantly, while hindquarter resistance did not decrease significantly in spinal rats. Mesenteric resistance returned to the control level with further bolus injection of Larginine (70 m g / k g iv), but renal and hindquarter resistances returned only partially. These results suggest that L-NAME induced hypertension is partially mediated by sympathetic vasoconstriction and the abnormal tones elicited in the superior mesenteric and renal areas are of spinal origin, while hindquarter tone may be of supraspinal origin, but is mediated also by a regional mechanism.
FREE COMMUNICATIONS 38-56; METHODOLOGY F38. A U T O M A T E D IMAGE P R O C E S S I N G SYSTEM F O R SHAPE RECOGNITION OF SINGLE RED BLOOD CEIJ.S BASED ON
OUT-OF-FOCUS
IMAGES
G. SCHNEIDER, G. ARTMANN AND G. H E N N I N G Department of Applied Cell Biophysics, FH Aachen, D-52429 Jfilich, Germany Shape recognition of red blood cells (RBC), including detection of subpopulations, is quite important in diagnosis. Apart from characteristic shape distribution due to different diseases, the shape response caused by (cell damaging) substances is a focus of interest. Automated procedures presently known can't classify single RBC, but only calculate an integral shape parameter of the whole blood sample. Additionally, calibration to a normal set of cells is required. Here, a technique is presented, which allows single classification of
238
Free Communications
Vol. 32, Nos. 2-3
RBC prepared on a special slide in a microscopic image processing system. Beside the standard requirements (light microscope, CCD-camera, frame grabber, standard PC) we used a self-made z-axis control for motor driven focusing and defocusing of the blood sample. Therefore, a new auto focus algorithm for microscopic phase objects based on geometrical optics theory was developed. In order to perform an obvious recognition, we acquire images from several focus planes and extract different geometric and intensity parameters (e.g., diameter, intensity from the cell border). The shape classes are similar to Bessis's definitions. However, echinocyte I-II and stomatocyte III have a continuous shape parameter allowing a substantially higher resolution of shape determination. As a conclusion, first results obtained with the shape changing substances chlorpromazine and sodium salicylate have shown that the computer based classification corresponds to those manually classified. In the near future, we want to investigate in which way certain diseases are associated with a characteristic shape distribution of RBCs in a sample.
F39. H I G H R E S O L U T I O N IMAGE C Y T O M E T R Y IN T H E ANALYSIS OF DIFFERENCES BETWEEN AA A N D SS B L O O D SPECIMENS L. WHEELESS, R. ROBINSON, V. MEYERS, O. LAPETS, P. ROWLEY, AND L. BENJAMIN Departments of Pathology and Laboratory Medicine and of Medicine, University of Rochester Medical Center, Rochester NY, 14642, and Montefiore Medical Center, Albert Einstein College of Medicine, Bronx, NY 10467, USA High resolution image cytometry provides quantitative data on the spatial distribution of hemoglobin within blood cells. Cell images are collected at 417 nm with a pixel resolution of 0.22 microns. A total of 58 metric, optical density, and textural features are derived from the digitized cell images. Textural features, Standard Deviation of Run Length Matrix Counts and Rotation Moment of the Cooccurrence Matrix, discriminate between patient mean values from AA (and AS) samples and those from SS (and SC) samples. This discrimination is made on normal appearing discoid (round) cells selected using the metric feature Form Factor. Antedating the classical morphological changes associated with sickling, this discrimination using textural features suggests that high resolution image cytometry may be an effective tool in the study and monitoring of sickle cell disease. Current studies utilize Factor Analysis to reduce dimensionality of the feature set while preserving information. Ten factors account for 88% of the variability in the data set of 5S features. Changes in patient means for these factors are observed as patients progress through crisis.
Free Communications
237
F37. INVOLVEMENT OF SYMPATHETIC VASOCONSTRICTION IN L-NAME INDUCED H Y P E R T E N S I O N IN C O N S C I O U S RATS N. IIDA Department of Physiology, School of Medicine, University of Hiroshima, Minarni-ku, Hiroshima 734, Japan The aims of the study are: (1) to d e t e r m i n e w h e t h e r sympathetic vasoconstriction is involved in L-NAME (inhibitor of NO synthase) induced hypertension in conscious rats, and (2) to understand whether or not the abnormal tone caused in peripheral vascular beds is of spinal origin. For these purposes, we studied the effects of ganglion blockade hexamethonium (C6) and L-arginine on L-NAME induced hypertension in intact and spinal rats. Spinal transection was performed at Thoracic 1 in ether-anesthetized rats with an electromagnetic flow probe placed around the superior mesenteric artery, renal artery or terminal aorta, and a catheter in the jugular vein for drug application. Peripheral resistance was calculated by dividing arterial pressure by peripheral blood flow. Three or 4 days after operation, regional blood flow and arterial pressure were measured in the conscious, resting state. L-NAME injection (3-5 m g / k g iv) resulted in a significant increase in mean arterial pressure and peripheral vascular resistances in both intact and spinal rats. Ten minutes after L-NAME injection, the infusion of C6 (25 m g / k g iv) reduced arterial pressure and peripheral vascular resistances significantly, while hindquarter resistance did not decrease significantly in spinal rats. Mesenteric resistance returned to the control level with further bolus injection of Larginine (70 m g / k g iv), but renal and hindquarter resistances returned only partially. These results suggest that L-NAME induced hypertension is partially mediated by sympathetic vasoconstriction and the abnormal tones elicited in the superior mesenteric and renal areas are of spinal origin, while hindquarter tone may be of supraspinal origin, but is mediated also by a regional mechanism.
FREE COMMUNICATIONS 38-56; METHODOLOGY F38. A U T O M A T E D IMAGE P R O C E S S I N G SYSTEM F O R SHAPE RECOGNITION OF SINGLE RED BLOOD CEIJ.S BASED ON
OUT-OF-FOCUS
IMAGES
G. SCHNEIDER, G. ARTMANN AND G. H E N N I N G Department of Applied Cell Biophysics, FH Aachen, D-52429 Jfilich, Germany Shape recognition of red blood cells (RBC), including detection of subpopulations, is quite important in diagnosis. Apart from characteristic shape distribution due to different diseases, the shape response caused by (cell damaging) substances is a focus of interest. Automated procedures presently known can't classify single RBC, but only calculate an integral shape parameter of the whole blood sample. Additionally, calibration to a normal set of cells is required. Here, a technique is presented, which allows single classification of
238
Free Communications
Vol. 32, Nos. 2-3
RBC prepared on a special slide in a microscopic image processing system. Beside the standard requirements (light microscope, CCD-camera, frame grabber, standard PC) we used a self-made z-axis control for motor driven focusing and defocusing of the blood sample. Therefore, a new auto focus algorithm for microscopic phase objects based on geometrical optics theory was developed. In order to perform an obvious recognition, we acquire images from several focus planes and extract different geometric and intensity parameters (e.g., diameter, intensity from the cell border). The shape classes are similar to Bessis's definitions. However, echinocyte I-II and stomatocyte III have a continuous shape parameter allowing a substantially higher resolution of shape determination. As a conclusion, first results obtained with the shape changing substances chlorpromazine and sodium salicylate have shown that the computer based classification corresponds to those manually classified. In the near future, we want to investigate in which way certain diseases are associated with a characteristic shape distribution of RBCs in a sample.
F39. H I G H R E S O L U T I O N IMAGE C Y T O M E T R Y IN T H E ANALYSIS OF DIFFERENCES BETWEEN AA A N D SS B L O O D SPECIMENS L. WHEELESS, R. ROBINSON, V. MEYERS, O. LAPETS, P. ROWLEY, AND L. BENJAMIN Departments of Pathology and Laboratory Medicine and of Medicine, University of Rochester Medical Center, Rochester NY, 14642, and Montefiore Medical Center, Albert Einstein College of Medicine, Bronx, NY 10467, USA High resolution image cytometry provides quantitative data on the spatial distribution of hemoglobin within blood cells. Cell images are collected at 417 nm with a pixel resolution of 0.22 microns. A total of 58 metric, optical density, and textural features are derived from the digitized cell images. Textural features, Standard Deviation of Run Length Matrix Counts and Rotation Moment of the Cooccurrence Matrix, discriminate between patient mean values from AA (and AS) samples and those from SS (and SC) samples. This discrimination is made on normal appearing discoid (round) cells selected using the metric feature Form Factor. Antedating the classical morphological changes associated with sickling, this discrimination using textural features suggests that high resolution image cytometry may be an effective tool in the study and monitoring of sickle cell disease. Current studies utilize Factor Analysis to reduce dimensionality of the feature set while preserving information. Ten factors account for 88% of the variability in the data set of 5S features. Changes in patient means for these factors are observed as patients progress through crisis.