PrintrLab incubator: A portable and low-cost CO2 incubator based on an open-source 3D printer architecture

Abstract

Growth in open-source hardware designs combined with the decreasing cost of high-quality 3D printers have supported a resurgence of in-house custom lab equipment development. Herein, we describe a low-cost (< $400), open-source CO2 incubator. The system is comprised of a Raspberry Pi computer connected to a 3D printer controller board that has controls for a CO2 sensor, solenoid valve, heater, and thermistors. CO2 is supplied through the sublimation of dry ice stored inside a thermos to create a sustained 5% CO2 supply. The unit is controlled via G-Code commands sent by the Raspberry Pi to the controller board. In addition, we built a custom software application for remote control and used the open-source Grafana dashboard for remote monitoring. Our data show that we can maintain consistent CO2 and temperature levels for over three days without manual interruption. The results from our culture plates and real-time PCR indicate that our incubator performed equally well when compared to a much more expensive commercial CO2 incubator. We have also demonstrated that the antibiotic susceptibility assay can be performed in this low-cost CO2 incubator. Our work also indicates that the system can be connected to incubator chambers of various chamber volumes.

A Stand-Alone Microfluidic Chip for Long-Term Cell Culture

“…To monitor in real time the conformation, mobility and proliferation of different cell lines, including 3T3, HepG2 and MB-231 cells, the fluidic chip was mounted on top of a microscope stage ( Figure 3 a). Our results demonstrate that with hourly medium exchange and constant 5% CO 2 exposure, the number of 3T3 fibroblasts nearly tripled within 48 h of incubation on the chip at both high and low seeding-cell density (0.05/cm 2 and 0.02/cm 2 , respectively) ( Figure 3 b–d), which is comparable to the growth rate in a conventional CO 2 incubator ( Figure S5 ) [ 23 , 24 , 25 ]. A similar growth rate, which is reflected by the cell number and counts of cell division, was observed for tumor-origin cell lines, i.e., MB and HepG2 cells ( Figure 4 ).…”

An Inexpensive Incubator for Mammalian Cell Culture Capable of Regulating O2, CO2, and Temperature

“…It weighs approximately one kilogram and as such is also highly portable within the lab. Certainly, it is not as portable as the CO 2 -alone incubators developed by Willbrand et al [13] or Arumagam et al [14]. These researchers focused on developing cell incubation systems suitable for fieldwork and/or shipping live cells.…”

“…It is capable of maintaining pre-set O 2 , CO 2 and temperature values within the physiological ranges for most mammalian cells. Although other "lab-assembled" O 2 monitoring systems [11], imaging systems [12], and cell culture incubators [13,14] exist, we are unaware of any that simultaneously regulate O 2 and CO 2 to enable experiments to be conducted at physiological O 2 levels. Thus, our incubator represents the least expensive solution to the important issue of maintaining physioxia in cell culture that is currently available.…”

Robotic System for Organoid Assembly in a Multi-Well Microfluidic Chip

“…While commercial miniature CO2 incubators exist that could fit in a hood, such as the MyTemp Mini CO2 Digital Incubator (Benchmark Scientific), these are not structurally modifiable, and therefore not appropriate for a prototyping system that might need to expand in size and be controlled in unconventional ways. Recently, several labs have constructed DIY CO2 incubators, [24][25][26][27][28] and the Pelling Lab implementation led to the company Incuvers that produces modifiable Arduinocontrolled incubators, 29 from which we acquired the Incuvers Model 1 (Figure 2A).…”