Dataset “STED_N2V_TestData.lif” is DNA distribution in a fixed zebrafish embryo, obtained with STED microscopy. It contains 6 series. And each series has low-quality images 1 and 2 which are acquired with identical acquisition settings and the high-quality image which was acquired in the same scanning sequence as the low-quality images but included accumulation by repeated line-scanning to improve image quality. Low-quality images 1 and 2 are then reconstructed by Noise2Void (Reconstructed image 1 and 2) and saved in the “reconstructed.zip” folder Python script "FRC.py" contains analysis to determine the improvement in the effective resolution of reconstructed images relative to the unprocessed low-quality images. Python script “n2v-SSIM-STED.py” illustrates how SSIM value improves by n2v processing on the dataset named “STED_N2V_TestData.lif