Handwriting is a written speech of an individual with characteristics peculiar to himself. The handwriting of a person is a product of his mental, emotional, intellectual and physical personality. Once the handwriting is set it is unique expression of his graphic personality. The differentiation for the identification of handedness of author plays an important role to identify an individual in document cases and to study this differentiation and comparison of left handed and right handed writers on the basis of strokes and slope of letter, the present study was conducted at Bilaspur district, Chhattisgarh in which 100 samples of handwriting, 50 left-handed and 50 right-handed writer were collected. The handwriting samples were examined by hand lens, flexible arm illuminated magnifier, grid scale and the characteristics features like stroke and slope were observed in various letters. A significant difference was observed in left handed and right handed writing and was concluded that left-handed writer make strokes in right-to-left direction and the slope of letters has an inclination in backward direction where as right-handed writer make strokes in left-to-right direction and the inclination of slope was in forward direction. Thus, this differentiation and comparison helps to solve many document cases.