Maturity at harvest determines seed viability and vigor. However, separating seeds from different stages of development can be difficult using existing seed sorting technologies. New technology non-destructively sorts seeds based on their chlorophyll fluorescence (CF), so seeds with the same dry weight but with different physiological maturates can be separated. We determined whether chlorophyll content of muskmelon (Cucumis melo L. cv. Top Mark) seeds changes during development and whether those changes were related to viability and vigor. Seed viability and vigor were determined using an Association of Official Seed Analysts wet paper towel germination test. `Top Mark' seeds from nine stages of development were run through the SeedMaster Analyzer (Satake USA Inc., Houston, Texas), which calculated the chlorophyll content of each seed. The CF signal was fed into a computer to obtain a frequency histogram. Forty, 45, and 55 days after anthesis (DAA) seeds had germination percentages of 96%, 98%, and 100%, respectively, the highest in the study. Fifty-five DAA had greater seed vigor and viability and contained the lowest CF values; 207 on the 1000-value scale. The less-mature seeds contained higher chlorophyll content and had the lowest seed vigor and viability. Seed vigor and chlorophyll content were negatively correlated in this study. All seeds with high CF values had low vigor, but not all seeds with low CF values have high vigor. Seed aging during storage can reduce viability and vigor independent of chlorophyll content. Based on chlorophyll content, the SeedMaster Analyzer can non-destructively remove immature, low-vigor seeds that have the same physical characteristics and weight as more mature seeds. Chlorophyll fluorescence technology may allow the seed industry to further improve seed quality and maximize vigor.