Morphological changes due to apoptosis induced by PBM alone and the combination with X-ray radiation were detected by fluorescence microscopy (Zeiss, Germany) of cultures stained with acridine orange/ethidium bromide (AO/EB) and quantified using flow cytometry according to published procedures [30 (link), 31 (link)]. Treated HeLa cells were pelleted, resuspended in 200 μL of PBS, and stained with AO-EB. The final concentrations of AO (Sigma, USA- A6014) and EB (Sigma, USA- E7637) were 0.1 and 0.25 mM, respectively. All samples were stained and analyzed immediately at room temperature. Flow cytometric analysis was performed on CyFlow Space (Parpec, Germany). Data were analyzed by using FloMax software.
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Apoptosis Imaging and Quantification
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Acridine orange
Apoptosis
Ethidium bromide
Flomax
Flow cytometric
Fluorescence microscopy
Hela cells
Radiation
X ray
Corresponding Organization : University of Tehran
Other organizations : Academic Center for Education, Culture and Research, Firoozgar General Hospital, Iran University of Medical Sciences, Massachusetts General Hospital, Harvard–MIT Division of Health Sciences and Technology, Harvard University
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Variable analysis
independent variables
- Photobiomodulation (PBM) alone
- Combination of PBM and X-ray radiation
- Morphological changes due to apoptosis
- Quantification of apoptosis using flow cytometry
- Culture conditions (e.g., temperature, media)
- Staining procedures (e.g., AO/EB concentrations, staining time)
- Positive control: Not explicitly mentioned
- Negative control: Not explicitly mentioned
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