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Trypan blue staining was performed as described previously, with minor modifications53. For staining, the samples were immersed in boiled lactophenol (glycerol:lactic acid:liquid phenol:distilled water, 1:1:1:1) with 0.25 mg/mL trypan blue for 20 s. Next, the samples were destained with destaining buffer (ethanol:liquid phenol:lactic acid, 2:1:1) at 65 °C for 1 h and washed three times with 75% (v/v) ethanol. Then, the samples were photographed under a microscope (MZ10F, Leica, Germany).
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