Awer betaine aldehyde applicaWon, CHCA matrix was applied with 10 layers and spraying nozzle posiWon Z=25 (intermediate posiWon above the Wssue). Matrix soluWon was made of 5mg/ml in ACN:H2O (1:1) and 0,2% of TFA. For NEDC matrix, 7mg/ml in 70% MeOH was prepared. Spraying nozzle posiWon has been set at Z=5 (high posiWon above the Wssue), and 10 layers were put on the glass. The flow rate for both matrices soluWons changed among layers: 15 µl/min for the first layer, 20 µl/min for the second layer, 30 µl/min for the third layer, 50 µl/min for the fourth layer and 60 µl/min for next layers. The spraying nozzle sprayed matrix soluWons linearly with line distance equal to 2mm and with a speed of 600 mm/min.