Quantification of gap junctional intercellular communication based on digital image analysis
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Quantification of gap junctional intercellular communication based on digital image analysis. / Hofgaard, Johannes P; Mollerup, Sarah; Holstein-Rathlou, Niels-Henrik; Nielsen, Morten Schak.
In: American Journal of Physiology: Regulatory, Integrative and Comparative Physiology, Vol. 297, No. 2, 2009, p. R243-7.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - Quantification of gap junctional intercellular communication based on digital image analysis
AU - Hofgaard, Johannes P
AU - Mollerup, Sarah
AU - Holstein-Rathlou, Niels-Henrik
AU - Nielsen, Morten Schak
N1 - Keywords: Animals; Carbenoxolone; Cell Communication; Cell Line, Tumor; Connexin 43; Connexins; Dextrans; Electroporation; Fluorescent Dyes; Gap Junctions; Image Processing, Computer-Assisted; Internet; Isoquinolines; Microscopy, Fluorescence; Neuroglia; Octanols; Rats; Software; Transfection
PY - 2009
Y1 - 2009
N2 - Intercellular communication via gap junction channels can be quantified by several methods based on diffusion of fluorescent dyes or metabolites. Given the variation in intercellular coupling of cells, even under untreated control conditions, it is of essence to quantify the coupling between numerous cells to obtain reliable estimates of metabolic coupling. Quantification is often based on manual counting of fluorescent cells, which is time consuming and may include some degree of subjectivity. In this report, we introduce a technique based on digital image analysis, and the software for the analysis is presented together with a detailed protocol in the online supplemental material (https://bmi.ku.dk/matlab_program/). Fluorescent dye was introduced in connexin 43-expressing C6 glioma cells by in situ electroporation, and fluorescence intensity was measured in the electroporated cells and in cells receiving dye by intercellular diffusion. The analysis performed is semiautomatic, and comparison with traditional cell counting shows that this method reliably determines the effect of uncoupling by several interventions. This new method of analysis yields a rapid and objective quantification process with a high degree of reproducibility.
AB - Intercellular communication via gap junction channels can be quantified by several methods based on diffusion of fluorescent dyes or metabolites. Given the variation in intercellular coupling of cells, even under untreated control conditions, it is of essence to quantify the coupling between numerous cells to obtain reliable estimates of metabolic coupling. Quantification is often based on manual counting of fluorescent cells, which is time consuming and may include some degree of subjectivity. In this report, we introduce a technique based on digital image analysis, and the software for the analysis is presented together with a detailed protocol in the online supplemental material (https://bmi.ku.dk/matlab_program/). Fluorescent dye was introduced in connexin 43-expressing C6 glioma cells by in situ electroporation, and fluorescence intensity was measured in the electroporated cells and in cells receiving dye by intercellular diffusion. The analysis performed is semiautomatic, and comparison with traditional cell counting shows that this method reliably determines the effect of uncoupling by several interventions. This new method of analysis yields a rapid and objective quantification process with a high degree of reproducibility.
U2 - 10.1152/ajpregu.00089.2009
DO - 10.1152/ajpregu.00089.2009
M3 - Journal article
C2 - 19535680
VL - 297
SP - R243-7
JO - American Journal of Physiology
JF - American Journal of Physiology
SN - 0363-6119
IS - 2
ER -
ID: 16213627