Air is an important part of human life. However, air can be contaminated with microorganisms such as airborne bacteria and fungi. Temperature and relative humidity in a room can have an impact on the quantity of airborne bacteria and fungi. This study aims to figure out the correlation between the number of airborne bacteria and fungi with temperature and relative humidity. In 15 rooms of Microbiology laboratory, NA and SDA Petri plates were placed, after incubation, the number of colonies in each plate was counted. Pearson test was conducted with SPSS to determine the correlation between temperature and relative humidity to the number of airborne bacteria and fungi. The highest number of airborne bacteria was in the reading room (352 CFU/m3), while the lowest number was in the laundry room (13 CFU/m3) and the highest number of airborne fungi was in the Mycology room (156 CFU/m3), while there were no airborne fungi found in the urine and laundry rooms. Based on the results of the Pearson test, it was found that the value of p = 0.668 (p> 0.5) showed that there was no correlation between temperature and the number of airborne bacteria and fungi. Based on the results of the Pearson test, the value of p = 0.745 (p> 0.5) showed that there was no correlation between relative humidity and the number of airborne bacteria and fungi. There is no correlation between temperature and relative humidity with the number of airborne bacteria and fungi.