# Microscope Enclosure for Temperature Regulation and Light Isolation
## 3 Results  
To assess temperature uniformity, we measured the temperature with and without the baffle panels at eight locations throughout the upright enclosure (Figure 2a). These measurements were conducted without the microscope in the enclosure. Temperature was monitored with the enclosure starting at room temperature (~20 \({}^{\circ}\)C) and the heater setpoint at 37 \({}^{\circ}\)C (AirTherm SMT, World Precisions Instruments). The measured temperature then increased over approximately 300 s, before stabilizing into oscillations (Figure 2b). These oscillations were attributed to heater feedback and could be altered by adjusting feedback parameters (the factory default parameters were used for these measurements). Without the baffle panels, we observed that the average temperature range across the eight probes was 5 \({}^{\circ}\)C. This average range was calculated by finding the difference between maximum and minimum probe temperature readings at each time point and then averaging over a time period when the enclosure temperature was near the 37 \({}^{\circ}\)C setpoint (500 s to 1000 s). We also observed that the highest temperature probe readings were near the air supply at the bottom of the enclosure (Figure 2b). When the input and output baffle panels were added, the average temperature range across the probes was also 5 \({}^{\circ}\)C (Figure 2c). Additionally, the baffles altered the temperature landscape, with higher temperatures occurring closer to the top of the enclosure. We attribute this difference to (1) the baffles distributing the air more uniformly across the entire height of the enclosure, and to (2) warm air rising within the box. The confluence of these together resulted in the top of the box being warmer than the bottom. In addition, the baffles also damped out the responsiveness of the temperature feedback, increasing the length of time for the temperature to initially equilibrate. The magnitude of the feedback oscillations also decreased. Specifically, for the enclosure without the baffles (Figure 2b), the average standard deviation of the eight probes was 0.5 \({}^{\circ}\)C, while the average standard deviation with the baffle panels (Figure 2c) was 0.3 \({}^{\circ}\)C.  
To account for the rising warm air in the enclosure, we closed the top half of the supply air baffle panel holes with tape. This resulted in a significant decrease in the average temperature range, with a new range of 2 \({}^{\circ}\)C between the probes (Figure 2d). Biasing the incoming air towards the bottom appeared to compensate for rising hot air. The average temperature standard devastation with respect to time also decreased to 0.2 \({}^{\circ}\)C. A measurement of temperature variation over a long duration, 48 h, was also acquired (Figure 2e). Temperature measurements were acquired every minute and the average temperature standard deviation of the eight probes was 0.1 \({}^{\circ}\)C. This measurement indicates the enclosure is stable for long time-lapse measurements. Next, we placed eight temperature probes at different locations in an enclosure with a microscope (Figure 3a). Like the previous experiment, we monitored the probes as the temperature increased from room temperature to 37 \({}^{\circ}\)C. In addition, the heater's temperature sensor was attached directly to the objective, as opposed to being in the air. In this experiment, we observed some of the probes went significantly beyond 37 \({}^{\circ}\)C, particularly those located on the air supply side (right side) of the enclosure or top of the enclosure. The probe mounted directly to the objective did not significantly overshoot the setpoint temperature. (Note: In this experiment, we used a less powerful, older generation heater (AirTherm, World Precision Instruments) and the enclosure volume was about twice the volume of the upright enclosure. Therefore, the equilibration period was significantly longer. )Figure 2: Temperature in the upright enclosure with and without baffle panels. (**a**) Thermocouples were placed at 8 locations in the enclosure.