Yıl 2018, Cilt 14, Sayı 1, Sayfalar 99 - 104 2018-03-30

Development of a Direct Trypan Blue Exclusion Method to Detect Cell Viability of Adherent Cells into ELISA Plates

Selcen Çelik Uzuner [1]

75 589

Cell viability detection is important in cell culture applications including measurement of cell proliferation i.e for understanding cytotoxic effects of compounds on cells. There are some cell viability methods based on fluorescence or non-fluorescence detection. More simplified evaluation for cell viability, such as trypan blue staining, can be preferred before performing fluorescence assays. This appears advantageous when to have a large number of cell samples in ELISA plates after treatments with different concentrations of drug candidates. Thus, further fluorescence assays can include less concentrations rather than experiencing all used along 96-well plates. For this, trypan blue exclusion method is an option. Traditionally, treated cells are harvested by centrifugation and incubated with trypan blue within tubes followed by transferring the mixture into a hemacytometer with two chambers and assessed under the microscope. Nevertheless, using a hemacytometer limits practicability of this method when analyzing various cell samples into 96-well plates at the same time. This study was aimed to adapt trypan blue method to in situ staining of adherent cells cultured on ELISA plates. For this, cells were fixed with different fixatives after trypan blue incubation to maintain cells in impenetrable meshwork, and paraformaldehyde was the most effective fixative. This modified protocol was validated by testing the effect of dimethylsulfoxide-a cytotoxic agent-on cells, and expectedly found that cell viability reduced with higher concentrations of dimethylsulfoxide suggesting that in situ detection of cell viability by trypan blue can be a useful tool for preliminary detection of cells cultured on ELISA plates before performing automatized experiments with such flow cytometer and/or microplate reader.

Cell viability, cell death, adherent cells, trypan blue exclusion, ELISA plate, fixation
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Orcid: 0000-0002-9558-7048
Yazar: Selcen Çelik Uzuner (Sorumlu Yazar)
Ülke: Turkey


Bibtex @araştırma makalesi { cbayarfbe372192, journal = {Celal Bayar Üniversitesi Fen Bilimleri Dergisi}, issn = {1305-130X}, eissn = {1305-1385}, address = {Celal Bayar Üniversitesi}, year = {2018}, volume = {14}, pages = {99 - 104}, doi = {10.18466/cbayarfbe.372192}, title = {Development of a Direct Trypan Blue Exclusion Method to Detect Cell Viability of Adherent Cells into ELISA Plates}, key = {cite}, author = {Çelik Uzuner, Selcen} }
APA Çelik Uzuner, S . (2018). Development of a Direct Trypan Blue Exclusion Method to Detect Cell Viability of Adherent Cells into ELISA Plates. Celal Bayar Üniversitesi Fen Bilimleri Dergisi, 14 (1), 99-104. DOI: 10.18466/cbayarfbe.372192
MLA Çelik Uzuner, S . "Development of a Direct Trypan Blue Exclusion Method to Detect Cell Viability of Adherent Cells into ELISA Plates". Celal Bayar Üniversitesi Fen Bilimleri Dergisi 14 (2018): 99-104 <http://dergipark.gov.tr/cbayarfbe/issue/36240/372192>
Chicago Çelik Uzuner, S . "Development of a Direct Trypan Blue Exclusion Method to Detect Cell Viability of Adherent Cells into ELISA Plates". Celal Bayar Üniversitesi Fen Bilimleri Dergisi 14 (2018): 99-104
RIS TY - JOUR T1 - Development of a Direct Trypan Blue Exclusion Method to Detect Cell Viability of Adherent Cells into ELISA Plates AU - Selcen Çelik Uzuner Y1 - 2018 PY - 2018 N1 - doi: 10.18466/cbayarfbe.372192 DO - 10.18466/cbayarfbe.372192 T2 - Celal Bayar Üniversitesi Fen Bilimleri Dergisi JF - Journal JO - JOR SP - 99 EP - 104 VL - 14 IS - 1 SN - 1305-130X-1305-1385 M3 - doi: 10.18466/cbayarfbe.372192 UR - http://dx.doi.org/10.18466/cbayarfbe.372192 Y2 - 2018 ER -
EndNote %0 Celal Bayar Üniversitesi Fen Bilimleri Dergisi Development of a Direct Trypan Blue Exclusion Method to Detect Cell Viability of Adherent Cells into ELISA Plates %A Selcen Çelik Uzuner %T Development of a Direct Trypan Blue Exclusion Method to Detect Cell Viability of Adherent Cells into ELISA Plates %D 2018 %J Celal Bayar Üniversitesi Fen Bilimleri Dergisi %P 1305-130X-1305-1385 %V 14 %N 1 %R doi: 10.18466/cbayarfbe.372192 %U 10.18466/cbayarfbe.372192
ISNAD Çelik Uzuner, Selcen . "Development of a Direct Trypan Blue Exclusion Method to Detect Cell Viability of Adherent Cells into ELISA Plates". Celal Bayar Üniversitesi Fen Bilimleri Dergisi 14 / 1 (Mart 2018): 99-104. http://dx.doi.org/10.18466/cbayarfbe.372192