Objective We explored the method and techniques of preparing frozen sections using different mice tissues as our experimental animal materials, to improve frozen section quality, and to increase the serviceability of these techniques for the non-clinical safety evaluation of monoclonal antibodies. Methods The heart, liver, spleen, lung, kidney, and brain tissues of healthy NIH mice were collected, embedded, frozen, and sliced using a Leica CM1950 thermostatic frozen microtome, and then stained by hematoxylin and eosin dye and sealed. Results The slices were observed by light microscopy, and the result show no fold, no knife marks, no damage, and no excessive ice crystals. The cell morphology was clear, with the nucleus and cytoplasm clearly colored, and staining was good. Conclusions In the process of making frozen slices, strict controls from the aspects of collecting, embedding, quick-freezing, temperature selection, and thermostatic frozen sectioning should be carried out. However, different tissues have differences according to their own characteristics, and should be treated differently. In addition, to prepare high-quality frozen sections, and to better serve the non-clinical safety evaluation of monoclonal antibodies, we collectively need a high sense of responsibility and patience while mastering key technical points.