Figure 3 | Scientific Reports

Figure 3

From: Assessing the tolerance to room temperature and viability of freeze-dried mice spermatozoa over long-term storage at room temperature under vacuum

Figure 3

Production of healthy offspring from FD spermatozoa stored at RT for 1 year. (a) Single spermatozoa were collected from Tesla-positive ampoules after rehydration. (b) Spermatozoa aggregates collected from Tesla-negative ampoules after rehydration. (c) Comparison of comet tail lengths of FD spermatozoa stored at RT for less than 1 week or 1 year. Asterisk indicates statistically significant differences between samples at P < 0.05. Zygotes derived from FD spermatozoa stored at RT for (d) 1 day or (e) 1 year and stained with anti-gamma-H2AX antibody. Images show male and female pronuclei stained with DAPI in blue (upper left), female pronuclei labeled with anti-H3K9 me2 antibody in green (upper right), gamma-H2AX signals indicating double-stranded DNA breaks in red (lower left), and merged images (lower right). (f) The brightness of each male pronucleus was plotted. The brightness of male pronucleus in (d) was 1.44 and in (e) was 1.50. Different letters indicate statistically significant differences at P < 0.05. (g) Live offspring derived from FD spermatozoa stored at RT for 1 year. (h) The rate of full-term development of embryos derived from FD spermatozoa preserved for 1 day, 3, 6, and 12 months at RT. The 1 day data and Fig. 2g, 1 day, Tesla Posi. is same. Different letters indicates statistically significant differences between samples (P < 0.05).

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