Quantification of apoptotic and lytic cell death by video microscopy in combination with artificial neural networks.
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Quantification of apoptotic and lytic cell death by video microscopy in combination with artificial neural networks. / Weisser, M; Tiegs, Gisa; Wendel, A; Uhlig, S.
In: Cytometry, Vol. 31, No. 1, 1, 1998, p. 20-28.Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review
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TY - JOUR
T1 - Quantification of apoptotic and lytic cell death by video microscopy in combination with artificial neural networks.
AU - Weisser, M
AU - Tiegs, Gisa
AU - Wendel, A
AU - Uhlig, S
PY - 1998
Y1 - 1998
N2 - Apoptosis is characterized by chromatin condensation and DNA fragmentation in the absence of release of cytosolic enzymes such as lactate dehydrogenase (LDH). In contrast, necrosis is characterized by cell swelling, membrane disintegration with cytosolic enzyme release, and absence of chromatin condensation. Staining of cells with Hoechst H33342 dye is a routine method for identifying apoptotic nuclei. However, this process is tedious and prone to individual bias. Therefore, we investigated the suitability of an artificial neural network (ANN) to recognize and distinguish between apoptosis and necrosis. Using a human endothelial cell line (ECV304), we trained an ANN with DNA-stained apoptotic and necrotic nuclei obtained from cells exposed for 8 h to cycloheximide (Chx; 100 microM)/tumour necrosis factor-alpha (TNF; 50 ng/ml) or tert-butylhydroperoxide (t-BH; 2 mM), respectively. After this training step, the ANN correctly assigned necrosis induced by t-BH and apoptosis induced via Chx/TNF, Chx/CD95 activation, Pseudomonas exotoxin A/TNF, or X-rays. In all cases, apoptosis and necrosis as assigned by the ANN correlated with DNA fragmentation and LDH release.
AB - Apoptosis is characterized by chromatin condensation and DNA fragmentation in the absence of release of cytosolic enzymes such as lactate dehydrogenase (LDH). In contrast, necrosis is characterized by cell swelling, membrane disintegration with cytosolic enzyme release, and absence of chromatin condensation. Staining of cells with Hoechst H33342 dye is a routine method for identifying apoptotic nuclei. However, this process is tedious and prone to individual bias. Therefore, we investigated the suitability of an artificial neural network (ANN) to recognize and distinguish between apoptosis and necrosis. Using a human endothelial cell line (ECV304), we trained an ANN with DNA-stained apoptotic and necrotic nuclei obtained from cells exposed for 8 h to cycloheximide (Chx; 100 microM)/tumour necrosis factor-alpha (TNF; 50 ng/ml) or tert-butylhydroperoxide (t-BH; 2 mM), respectively. After this training step, the ANN correctly assigned necrosis induced by t-BH and apoptosis induced via Chx/TNF, Chx/CD95 activation, Pseudomonas exotoxin A/TNF, or X-rays. In all cases, apoptosis and necrosis as assigned by the ANN correlated with DNA fragmentation and LDH release.
KW - Humans
KW - Cell Line
KW - Microscopy, Video
KW - Necrosis
KW - Apoptosis/physiology
KW - Cell Death/physiology
KW - Cell Nucleus/physiology
KW - Discriminant Analysis
KW - Neural Networks (Computer)
KW - Humans
KW - Cell Line
KW - Microscopy, Video
KW - Necrosis
KW - Apoptosis/physiology
KW - Cell Death/physiology
KW - Cell Nucleus/physiology
KW - Discriminant Analysis
KW - Neural Networks (Computer)
M3 - SCORING: Journal article
VL - 31
SP - 20
EP - 28
IS - 1
M1 - 1
ER -
