The objective of this research work was to develop a very quick method for cotton fiber measurements. The importance of the work was to establish an objective fast and inexpensive method of determining the cotton fiber maturity. Cotton fiber maturity is generally accepted as measurement of the fiber wall, which has developed to an acceptable level of thickening. The measurements of the basic characteristics for the assessment of the maturity were collected and calculated by using software specifically developed for cotton maturity testing using Image Analyzer. The automated measuring system by image analyzer using new developed sample preparation procedure has the feasibility of providing with 500-1000 fiber cross-section data in very short time. Image analysis software is developed to measure following characteristics: Whole area of the fiber, Lumen, Perimeter & Diameter (ribbon width) and from collected data to calculate factors for the maturity evaluation. Calculated factors for the maturity data are: Wall thickness, Circularity, Degree of thickening, Maturity ratio, Relative Maturity & Maturity factor. Image Analysis method of cotton maturity measurements will provide cotton producers and users with precise, fast and accurate data of cotton fiber quality with the feasibility of adding the image analyzer to the HVI testing equipment.