The invention relates to an adhesive composition for coating of a glass slide and a single-surface coating technology. The adhesive composition is prepared from the following components according to amass / volume ratio: a component 1 and a component 2, wherein the component 1 is prepared from 0.05 to 0.3% of film forming agent (polyvinyl alcohol), 0.02 to 0.25% of adhesive (carboxymethylcellulose), and the balance of water; the component 2 is prepared from 0.2 to 1mg / 100ml of cationic swelling agent (polylysine), and the balance of water. The single-surface coating technology comprises the following steps of cleaning the glass slide by ultrasonic waves in an alkaline solution, drying, spraying and coating the component 1 to the front surface according to the amount of 20 to 50mu l per piece, drying at the temperature of 35 to 55 DEG C, cooling to room temperature, spraying and coating the component 2 to the same surface according to the amount of 10 to 30mu l per piece, drying at the temperature of 35 to 55 DEG C, and packaging under the vacuum condition. The adhesive composition has the advantages that the effects of high adhesivity, uniform distribution and high definition of cells or tissues on the glass slide are realized; the continuous, automatic, stable, uniform and batch type single-surface zonal spraying and coating technology is realized. The prepared pathology slidecan resist the repeated washing and soaking by various alcohols and reagents, be suitable for the wall attaching, single-layer distribution and no falling of the cells, the tissues and the like, and can reach the effects of clear background and no impurity.