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An Inexpensive 'Homemade' O2/CO2 Regulating Incubator for Mammalian Cell Culture

Our incubator is an inexpensive, portable and user friendly device that is capable of regulating O2, CO2 and temperature within physiological ranges. Using an Arduino-based approach to control O2 and CO2 headspace levels via N2 and CO2 gas injection, we were able to convert a temperature-regulating Styrofoam egg incubator into a efficient benchtop cell culture tool comparable to commercial laboratory O2/CO2 incubators. This incubator can be assembled for less than $1000 (CAD), making it highly accessable for biologists seeking to transition towards physiological cell culture practices.

Table of Contents

General Information

  • Culturing cells under physiologically-relevant conditions is vital to maintaining cellular phenotypes representative of in vivo biology. While O2 constitutes ~20% of the atmosphere, it's level ranges from 2 to 6% in most mammalian tissues, thus O2/CO2 incubators capable of establishing such conditions are necessary. Unfortuanately, commercial O2/CO2 incubators suitable for mammalian cell culture cost well over CAD $10,000, making them inaccessible to many cell biologists.
  • The "Homemade" O2/CO2 incubator described here works to address this obstacle by providing cell culturists with the means to self assemble an effective O2/CO2 incubator from a standard temperature-regulating Styrofoam egg incubator.
  • The Stuart lab is a large proponent of the importance of physiological conditions in cell culture, thus by making tools to maintain physiological oxygen levels more readily available, we hope to encourage the field to incorporate such practices into standard workflow.

Project Road Map

  • 📖 [The Publication](Coming Soon) - This incubator was published in an international, peer-reviewed, open-access journal called Oxygen. All original designs and assembly details can be found in this manuscript + the supplementary data sections.
  • 💻 Arduino Code
  • 🛒 [Materials List] (Coming Soon)
  • ❓ Issue Forum - Submit questions, concerns and assembly requests here!

Technologies Used

Design

Initial PrototypePicture2

Final Model: gas sensors, arduino controller, soleniod valves and electronics enclosed within a 3D-printed boxPicture3

Setup

Usage

  • This benchtop incubator is primarily intended for academic research labs looking to incorporate physioxic cell culture conditions. The interior of the incubator is suitable for both adherent and suspension cell culture, comfortably accomodating at least 25 100mm stacked tissue culture disks and/or flasks.
  • Specific setpoint gas levels (O2 and CO2) can be altered to suit the conditions required by the user. The code is currently set to maintain a physioxic level of 5% O2, and standard 5% CO2.

Points to Keep in Mind

  • Depending on the application, the Uno and solenoid valves can be powered in one of two ways. For applications where the system is only run for short periods of time, both the Uno and solenoids can be powered with a single 12VDC power supply. However, if the application requires the Uno to be constantly powered, as in the incubator application developed here, the Uno should be powered with a lower voltage (9VDC), while the solenoid valves are powered independently with a 12VDC power supply.
  • Heatsinks may be required for various components including the transistors, though we did not find them necessary under our conditions. Monitoring the temperature of electrical components is critical in the early stages of development to determine the need for heatsinks.

Contact

Created by @StuartLab -- Feel free to contact us with questions or concerns about incubator assembly!

About

O2/CO2 incubators for cell culture

Resources

Readme
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3 stars

Watchers

2 watching

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, 'i'); if (__m === '*' || __re.test(location.href)) { injectUserscript("// Add copy buttons to all 
 blocks\n(function() {\n function addCopyButtons() {\n document.querySelectorAll('pre code').forEach(function(codeBlock) {\n if (codeBlock.parentElement.hasAttribute('data-copy-added')) return;\n codeBlock.parentElement.setAttribute('data-copy-added', 'true');\n \n var btn = document.createElement('button');\n btn.textContent = 'Copy';\n btn.style.cssText = 'position:absolute;top:4px;right:4px;padding:2px 8px;font-size:11px;background:#4ecdc4;border:none;border-radius:4px;color:#1a1a2e;cursor:pointer;opacity:0.7;transition:opacity 0.2s;';\n btn.onmouseover = function() { this.style.opacity = '1'; };\n btn.onmouseout = function() { this.style.opacity = '0.7'; };\n btn.onclick = function() {\n navigator.clipboard.writeText(codeBlock.textContent).then(function() {\n btn.textContent = 'Copied!';\n setTimeout(function() { btn.textContent = 'Copy'; }, 1500);\n });\n };\n codeBlock.parentElement.style.position = 'relative';\n codeBlock.parentElement.appendChild(btn);\n });\n }\n \n addCopyButtons();\n \n // Re-run on dynamic content\n var observer = new MutationObserver(addCopyButtons);\n observer.observe(document.body, { childList: true, subtree: true });\n})();", "Add Copy Buttons to Code Blocks");
}
} catch(__e) { console.warn('[Userscript:Add Copy Buttons to Code Blocks]', __e); }
})();
(function(){
try {
var __m = "github.com";
var __re = new RegExp('^' + "github\\.com" + '

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An Inexpensive 'Homemade' O2/CO2 Regulating Incubator for Mammalian Cell Culture
Our incubator is an inexpensive, portable and user friendly device that is capable of regulating O2, CO2 and temperature within physiological ranges. Using an Arduino-based approach to control O2 and CO2 headspace levels via N2 and CO2 gas injection, we were able to convert a temperature-regulating Styrofoam egg incubator into a efficient benchtop cell culture tool comparable to commercial laboratory O2/CO2 incubators. This incubator can be assembled for less than $1000 (CAD), making it highly accessable for biologists seeking to transition towards physiological cell culture practices.
Table of Contents
General Information
  • Culturing cells under physiologically-relevant conditions is vital to maintaining cellular phenotypes representative of in vivo biology. While O2 constitutes ~20% of the atmosphere, it's level ranges from 2 to 6% in most mammalian tissues, thus O2/CO2 incubators capable of establishing such conditions are necessary. Unfortuanately, commercial O2/CO2 incubators suitable for mammalian cell culture cost well over CAD $10,000, making them inaccessible to many cell biologists.
  • The "Homemade" O2/CO2 incubator described here works to address this obstacle by providing cell culturists with the means to self assemble an effective O2/CO2 incubator from a standard temperature-regulating Styrofoam egg incubator.
  • The Stuart lab is a large proponent of the importance of physiological conditions in cell culture, thus by making tools to maintain physiological oxygen levels more readily available, we hope to encourage the field to incorporate such practices into standard workflow.
Project Road Map
  • 📖 [The Publication](Coming Soon) - This incubator was published in an international, peer-reviewed, open-access journal called Oxygen. All original designs and assembly details can be found in this manuscript + the supplementary data sections.
  • 💻 Arduino Code
  • 🛒 [Materials List] (Coming Soon)
  • ❓ Issue Forum - Submit questions, concerns and assembly requests here!
Technologies Used
Design
Initial PrototypePicture2
Final Model: gas sensors, arduino controller, soleniod valves and electronics enclosed within a 3D-printed boxPicture3
Setup
Usage
  • This benchtop incubator is primarily intended for academic research labs looking to incorporate physioxic cell culture conditions. The interior of the incubator is suitable for both adherent and suspension cell culture, comfortably accomodating at least 25 100mm stacked tissue culture disks and/or flasks.
  • Specific setpoint gas levels (O2 and CO2) can be altered to suit the conditions required by the user. The code is currently set to maintain a physioxic level of 5% O2, and standard 5% CO2.
Points to Keep in Mind
  • Depending on the application, the Uno and solenoid valves can be powered in one of two ways. For applications where the system is only run for short periods of time, both the Uno and solenoids can be powered with a single 12VDC power supply. However, if the application requires the Uno to be constantly powered, as in the incubator application developed here, the Uno should be powered with a lower voltage (9VDC), while the solenoid valves are powered independently with a 12VDC power supply.
  • Heatsinks may be required for various components including the transistors, though we did not find them necessary under our conditions. Monitoring the temperature of electrical components is critical in the early stages of development to determine the need for heatsinks.
Contact
Created by @StuartLab -- Feel free to contact us with questions or concerns about incubator assembly!
About
O2/CO2 incubators for cell culture
Resources
Readme
Activity
Stars
3 stars
Watchers
2 watching
Forks
1 fork
Report repository
Releases
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Contributors

, 'i');
if (__m === '*' || __re.test(location.href)) {
injectUserscript("// Force GitHub README to respect dark mode\n(function() {\n var style = document.createElement('style');\n style.textContent = '\n .markdown-body {\n color-scheme: dark light;\n }\n .markdown-body pre { background: #161b22 !important; }\n .markdown-body code { background: rgba(110, 118, 129, 0.4) !important; }\n .markdown-body table th, .markdown-body table td { border-color: #30363d !important; }\n .markdown-body img { background: #0d1117; }\n .markdown-body blockquote { border-left-color: #8b949e; }\n .markdown-body hr { border-color: #30363d; }\n ';\n document.head.appendChild(style);\n})();", "GitHub Dark Mode README Fix");
}
} catch(__e) { console.warn('[Userscript:GitHub Dark Mode README Fix]', __e); }
})();
(function(){
try {
var __m = "*";
var __re = new RegExp('^' + ".*" + '

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An Inexpensive 'Homemade' O2/CO2 Regulating Incubator for Mammalian Cell Culture
Our incubator is an inexpensive, portable and user friendly device that is capable of regulating O2, CO2 and temperature within physiological ranges. Using an Arduino-based approach to control O2 and CO2 headspace levels via N2 and CO2 gas injection, we were able to convert a temperature-regulating Styrofoam egg incubator into a efficient benchtop cell culture tool comparable to commercial laboratory O2/CO2 incubators. This incubator can be assembled for less than $1000 (CAD), making it highly accessable for biologists seeking to transition towards physiological cell culture practices.
Table of Contents
General Information
  • Culturing cells under physiologically-relevant conditions is vital to maintaining cellular phenotypes representative of in vivo biology. While O2 constitutes ~20% of the atmosphere, it's level ranges from 2 to 6% in most mammalian tissues, thus O2/CO2 incubators capable of establishing such conditions are necessary. Unfortuanately, commercial O2/CO2 incubators suitable for mammalian cell culture cost well over CAD $10,000, making them inaccessible to many cell biologists.
  • The "Homemade" O2/CO2 incubator described here works to address this obstacle by providing cell culturists with the means to self assemble an effective O2/CO2 incubator from a standard temperature-regulating Styrofoam egg incubator.
  • The Stuart lab is a large proponent of the importance of physiological conditions in cell culture, thus by making tools to maintain physiological oxygen levels more readily available, we hope to encourage the field to incorporate such practices into standard workflow.
Project Road Map
  • 📖 [The Publication](Coming Soon) - This incubator was published in an international, peer-reviewed, open-access journal called Oxygen. All original designs and assembly details can be found in this manuscript + the supplementary data sections.
  • 💻 Arduino Code
  • 🛒 [Materials List] (Coming Soon)
  • ❓ Issue Forum - Submit questions, concerns and assembly requests here!
Technologies Used
Design
Initial PrototypePicture2
Final Model: gas sensors, arduino controller, soleniod valves and electronics enclosed within a 3D-printed boxPicture3
Setup
Usage
  • This benchtop incubator is primarily intended for academic research labs looking to incorporate physioxic cell culture conditions. The interior of the incubator is suitable for both adherent and suspension cell culture, comfortably accomodating at least 25 100mm stacked tissue culture disks and/or flasks.
  • Specific setpoint gas levels (O2 and CO2) can be altered to suit the conditions required by the user. The code is currently set to maintain a physioxic level of 5% O2, and standard 5% CO2.
Points to Keep in Mind
  • Depending on the application, the Uno and solenoid valves can be powered in one of two ways. For applications where the system is only run for short periods of time, both the Uno and solenoids can be powered with a single 12VDC power supply. However, if the application requires the Uno to be constantly powered, as in the incubator application developed here, the Uno should be powered with a lower voltage (9VDC), while the solenoid valves are powered independently with a 12VDC power supply.
  • Heatsinks may be required for various components including the transistors, though we did not find them necessary under our conditions. Monitoring the temperature of electrical components is critical in the early stages of development to determine the need for heatsinks.
Contact
Created by @StuartLab -- Feel free to contact us with questions or concerns about incubator assembly!
About
O2/CO2 incubators for cell culture
Resources
Readme
Activity
Stars
3 stars
Watchers
2 watching
Forks
1 fork
Report repository
Releases
Packages
Contributors

, 'i');
if (__m === '*' || __re.test(location.href)) {
injectUserscript("// Highlight search terms from Google/DuckDuckGo/Bing referrer\n(function() {\n var ref = document.referrer;\n var terms = [];\n \n if (ref.includes('google.com') || ref.includes('duckduckgo.com') || ref.includes('bing.com')) {\n var url = new URL(ref);\n var q = url.searchParams.get('q') || url.searchParams.get('p');\n if (q) {\n terms = q.split(/\\s+/).filter(function(t) { return t.length > 2; });\n }\n }\n \n if (terms.length === 0) return;\n \n var style = document.createElement('style');\n style.textContent = '.userscript-highlight { background: #fbbf24; color: #1a1a2e; padding: 1px 3px; border-radius: 2px; }';\n document.head.appendChild(style);\n \n function highlight(node) {\n if (node.nodeType === 3) { // text node\n var text = node.textContent;\n var found = false;\n terms.forEach(function(term) {\n var regex = new RegExp('(' + term.replace(/[.*+?^${}()|[\\]\\\\]/g, '\\\\') + ')', 'gi');\n if (regex.test(text)) {\n found = true;\n var frag = document.createDocumentFragment();\n var parts = text.split(regex);\n parts.forEach(function(part, i) {\n if (i % 2 === 0) {\n frag.appendChild(document.createTextNode(part));\n } else {\n var span = document.createElement('span');\n span.className = 'userscript-highlight';\n span.textContent = part;\n frag.appendChild(span);\n }\n });\n node.parentNode.replaceChild(frag, node);\n }\n });\n } else if (node.nodeType === 1 && node.childNodes) { // element\n var skipTags = ['SCRIPT', 'STYLE', 'NOSCRIPT', 'TEXTAREA', 'INPUT', 'SELECT'];\n if (!skipTags.includes(node.tagName)) {\n Array.from(node.childNodes).forEach(highlight);\n }\n }\n }\n \n highlight(document.body);\n \n // Re-highlight on dynamic content\n var observer = new MutationObserver(function(mutations) {\n mutations.forEach(function(m) {\n m.addedNodes.forEach(function(node) {\n if (node.nodeType === 1 || node.nodeType === 3) highlight(node);\n });\n });\n });\n observer.observe(document.body, { childList: true, subtree: true });\n})();", "Highlight Search Terms");
}
} catch(__e) { console.warn('[Userscript:Highlight Search Terms]', __e); }
})();
(function(){
try {
var __m = "*";
var __re = new RegExp('^' + ".*" + '

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An Inexpensive 'Homemade' O2/CO2 Regulating Incubator for Mammalian Cell Culture
Our incubator is an inexpensive, portable and user friendly device that is capable of regulating O2, CO2 and temperature within physiological ranges. Using an Arduino-based approach to control O2 and CO2 headspace levels via N2 and CO2 gas injection, we were able to convert a temperature-regulating Styrofoam egg incubator into a efficient benchtop cell culture tool comparable to commercial laboratory O2/CO2 incubators. This incubator can be assembled for less than $1000 (CAD), making it highly accessable for biologists seeking to transition towards physiological cell culture practices.
Table of Contents
General Information
  • Culturing cells under physiologically-relevant conditions is vital to maintaining cellular phenotypes representative of in vivo biology. While O2 constitutes ~20% of the atmosphere, it's level ranges from 2 to 6% in most mammalian tissues, thus O2/CO2 incubators capable of establishing such conditions are necessary. Unfortuanately, commercial O2/CO2 incubators suitable for mammalian cell culture cost well over CAD $10,000, making them inaccessible to many cell biologists.
  • The "Homemade" O2/CO2 incubator described here works to address this obstacle by providing cell culturists with the means to self assemble an effective O2/CO2 incubator from a standard temperature-regulating Styrofoam egg incubator.
  • The Stuart lab is a large proponent of the importance of physiological conditions in cell culture, thus by making tools to maintain physiological oxygen levels more readily available, we hope to encourage the field to incorporate such practices into standard workflow.
Project Road Map
  • 📖 [The Publication](Coming Soon) - This incubator was published in an international, peer-reviewed, open-access journal called Oxygen. All original designs and assembly details can be found in this manuscript + the supplementary data sections.
  • 💻 Arduino Code
  • 🛒 [Materials List] (Coming Soon)
  • ❓ Issue Forum - Submit questions, concerns and assembly requests here!
Technologies Used
Design
Initial PrototypePicture2
Final Model: gas sensors, arduino controller, soleniod valves and electronics enclosed within a 3D-printed boxPicture3
Setup
Usage
  • This benchtop incubator is primarily intended for academic research labs looking to incorporate physioxic cell culture conditions. The interior of the incubator is suitable for both adherent and suspension cell culture, comfortably accomodating at least 25 100mm stacked tissue culture disks and/or flasks.
  • Specific setpoint gas levels (O2 and CO2) can be altered to suit the conditions required by the user. The code is currently set to maintain a physioxic level of 5% O2, and standard 5% CO2.
Points to Keep in Mind
  • Depending on the application, the Uno and solenoid valves can be powered in one of two ways. For applications where the system is only run for short periods of time, both the Uno and solenoids can be powered with a single 12VDC power supply. However, if the application requires the Uno to be constantly powered, as in the incubator application developed here, the Uno should be powered with a lower voltage (9VDC), while the solenoid valves are powered independently with a 12VDC power supply.
  • Heatsinks may be required for various components including the transistors, though we did not find them necessary under our conditions. Monitoring the temperature of electrical components is critical in the early stages of development to determine the need for heatsinks.
Contact
Created by @StuartLab -- Feel free to contact us with questions or concerns about incubator assembly!
About
O2/CO2 incubators for cell culture
Resources
Readme
Activity
Stars
3 stars
Watchers
2 watching
Forks
1 fork
Report repository
Releases
Packages
Contributors

, 'i');
if (__m === '*' || __re.test(location.href)) {
injectUserscript("// Strip utm_, fbclid, gclid, etc. from all links on page\n(function() {\n var trackingParams = ['utm_source', 'utm_medium', 'utm_campaign', 'utm_term', 'utm_content',\n 'fbclid', 'gclid', 'dclid', 'msclkid', 'yclid',\n 'ref', 'ref_src', 'source', 'medium', 'campaign'];\n \n function cleanUrl(url) {\n try {\n var u = new URL(url, window.location.origin);\n var changed = false;\n trackingParams.forEach(function(p) {\n if (u.searchParams.has(p)) {\n u.searchParams.delete(p);\n changed = true;\n }\n });\n return changed ? u.toString() : url;\n } catch (e) {\n return url;\n }\n }\n \n function cleanLinks() {\n document.querySelectorAll('a[href]').forEach(function(a) {\n var clean = cleanUrl(a.href);\n if (clean !== a.href) a.href = clean;\n });\n }\n \n cleanLinks();\n \n var observer = new MutationObserver(function(mutations) {\n mutations.forEach(function(m) {\n m.addedNodes.forEach(function(node) {\n if (node.nodeType === 1) {\n if (node.tagName === 'A') cleanLinks();\n node.querySelectorAll('a[href]').forEach(function(a) {\n var clean = cleanUrl(a.href);\n if (clean !== a.href) a.href = clean;\n });\n }\n });\n });\n });\n observer.observe(document.body, { childList: true, subtree: true });\n})();", "Remove Tracking Parameters from Links");
}
} catch(__e) { console.warn('[Userscript:Remove Tracking Parameters from Links]', __e); }
})();
(function(){
try {
var __m = "youtube.com";
var __re = new RegExp('^' + "youtube\\.com" + '

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An Inexpensive 'Homemade' O2/CO2 Regulating Incubator for Mammalian Cell Culture
Our incubator is an inexpensive, portable and user friendly device that is capable of regulating O2, CO2 and temperature within physiological ranges. Using an Arduino-based approach to control O2 and CO2 headspace levels via N2 and CO2 gas injection, we were able to convert a temperature-regulating Styrofoam egg incubator into a efficient benchtop cell culture tool comparable to commercial laboratory O2/CO2 incubators. This incubator can be assembled for less than $1000 (CAD), making it highly accessable for biologists seeking to transition towards physiological cell culture practices.
Table of Contents
General Information
  • Culturing cells under physiologically-relevant conditions is vital to maintaining cellular phenotypes representative of in vivo biology. While O2 constitutes ~20% of the atmosphere, it's level ranges from 2 to 6% in most mammalian tissues, thus O2/CO2 incubators capable of establishing such conditions are necessary. Unfortuanately, commercial O2/CO2 incubators suitable for mammalian cell culture cost well over CAD $10,000, making them inaccessible to many cell biologists.
  • The "Homemade" O2/CO2 incubator described here works to address this obstacle by providing cell culturists with the means to self assemble an effective O2/CO2 incubator from a standard temperature-regulating Styrofoam egg incubator.
  • The Stuart lab is a large proponent of the importance of physiological conditions in cell culture, thus by making tools to maintain physiological oxygen levels more readily available, we hope to encourage the field to incorporate such practices into standard workflow.
Project Road Map
  • 📖 [The Publication](Coming Soon) - This incubator was published in an international, peer-reviewed, open-access journal called Oxygen. All original designs and assembly details can be found in this manuscript + the supplementary data sections.
  • 💻 Arduino Code
  • 🛒 [Materials List] (Coming Soon)
  • ❓ Issue Forum - Submit questions, concerns and assembly requests here!
Technologies Used
Design
Initial PrototypePicture2
Final Model: gas sensors, arduino controller, soleniod valves and electronics enclosed within a 3D-printed boxPicture3
Setup
Usage
  • This benchtop incubator is primarily intended for academic research labs looking to incorporate physioxic cell culture conditions. The interior of the incubator is suitable for both adherent and suspension cell culture, comfortably accomodating at least 25 100mm stacked tissue culture disks and/or flasks.
  • Specific setpoint gas levels (O2 and CO2) can be altered to suit the conditions required by the user. The code is currently set to maintain a physioxic level of 5% O2, and standard 5% CO2.
Points to Keep in Mind
  • Depending on the application, the Uno and solenoid valves can be powered in one of two ways. For applications where the system is only run for short periods of time, both the Uno and solenoids can be powered with a single 12VDC power supply. However, if the application requires the Uno to be constantly powered, as in the incubator application developed here, the Uno should be powered with a lower voltage (9VDC), while the solenoid valves are powered independently with a 12VDC power supply.
  • Heatsinks may be required for various components including the transistors, though we did not find them necessary under our conditions. Monitoring the temperature of electrical components is critical in the early stages of development to determine the need for heatsinks.
Contact
Created by @StuartLab -- Feel free to contact us with questions or concerns about incubator assembly!
About
O2/CO2 incubators for cell culture
Resources
Readme
Activity
Stars
3 stars
Watchers
2 watching
Forks
1 fork
Report repository
Releases
Packages
Contributors

, 'i');
if (__m === '*' || __re.test(location.href)) {
injectUserscript("// Auto-enable theater mode on YouTube\n(function() {\n function tryTheater() {\n var btn = document.querySelector('button[aria-label=\"Theater mode\"], ytd-player #player button[title=\"Theater mode\"]');\n if (btn && !btn.classList.contains('activated')) {\n btn.click();\n }\n }\n \n // Try immediately\n tryTheater();\n \n // Try after navigation (SPA)\n var lastUrl = location.href;\n setInterval(function() {\n if (location.href !== lastUrl) {\n lastUrl = location.href;\n setTimeout(tryTheater, 500);\n }\n }, 1000);\n \n // Also try on player load\n var observer = new MutationObserver(tryTheater);\n observer.observe(document.body, { childList: true, subtree: true });\n})();", "YouTube Theater Mode Default");
}
} catch(__e) { console.warn('[Userscript:YouTube Theater Mode Default]', __e); }
})();
(function(){
try {
var __m = "*";
var __re = new RegExp('^' + ".*" + '

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An Inexpensive 'Homemade' O2/CO2 Regulating Incubator for Mammalian Cell Culture
Our incubator is an inexpensive, portable and user friendly device that is capable of regulating O2, CO2 and temperature within physiological ranges. Using an Arduino-based approach to control O2 and CO2 headspace levels via N2 and CO2 gas injection, we were able to convert a temperature-regulating Styrofoam egg incubator into a efficient benchtop cell culture tool comparable to commercial laboratory O2/CO2 incubators. This incubator can be assembled for less than $1000 (CAD), making it highly accessable for biologists seeking to transition towards physiological cell culture practices.
Table of Contents
General Information
  • Culturing cells under physiologically-relevant conditions is vital to maintaining cellular phenotypes representative of in vivo biology. While O2 constitutes ~20% of the atmosphere, it's level ranges from 2 to 6% in most mammalian tissues, thus O2/CO2 incubators capable of establishing such conditions are necessary. Unfortuanately, commercial O2/CO2 incubators suitable for mammalian cell culture cost well over CAD $10,000, making them inaccessible to many cell biologists.
  • The "Homemade" O2/CO2 incubator described here works to address this obstacle by providing cell culturists with the means to self assemble an effective O2/CO2 incubator from a standard temperature-regulating Styrofoam egg incubator.
  • The Stuart lab is a large proponent of the importance of physiological conditions in cell culture, thus by making tools to maintain physiological oxygen levels more readily available, we hope to encourage the field to incorporate such practices into standard workflow.
Project Road Map
  • 📖 [The Publication](Coming Soon) - This incubator was published in an international, peer-reviewed, open-access journal called Oxygen. All original designs and assembly details can be found in this manuscript + the supplementary data sections.
  • 💻 Arduino Code
  • 🛒 [Materials List] (Coming Soon)
  • ❓ Issue Forum - Submit questions, concerns and assembly requests here!
Technologies Used
Design
Initial PrototypePicture2
Final Model: gas sensors, arduino controller, soleniod valves and electronics enclosed within a 3D-printed boxPicture3
Setup
Usage
  • This benchtop incubator is primarily intended for academic research labs looking to incorporate physioxic cell culture conditions. The interior of the incubator is suitable for both adherent and suspension cell culture, comfortably accomodating at least 25 100mm stacked tissue culture disks and/or flasks.
  • Specific setpoint gas levels (O2 and CO2) can be altered to suit the conditions required by the user. The code is currently set to maintain a physioxic level of 5% O2, and standard 5% CO2.
Points to Keep in Mind
  • Depending on the application, the Uno and solenoid valves can be powered in one of two ways. For applications where the system is only run for short periods of time, both the Uno and solenoids can be powered with a single 12VDC power supply. However, if the application requires the Uno to be constantly powered, as in the incubator application developed here, the Uno should be powered with a lower voltage (9VDC), while the solenoid valves are powered independently with a 12VDC power supply.
  • Heatsinks may be required for various components including the transistors, though we did not find them necessary under our conditions. Monitoring the temperature of electrical components is critical in the early stages of development to determine the need for heatsinks.
Contact
Created by @StuartLab -- Feel free to contact us with questions or concerns about incubator assembly!
About
O2/CO2 incubators for cell culture
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, 'i');
if (__m === '*' || __re.test(location.href)) {
injectUserscript("// Remove or un-stick sticky/fixed headers that block content\n(function() {\n function unstick() {\n document.querySelectorAll('header, nav, [role=\"banner\"], .header, .navbar, .sticky, .fixed-top, [style*=\"position: fixed\"], [style*=\"position:sticky\"]').forEach(function(el) {\n if (el.style.position === 'fixed' || el.style.position === 'sticky' || \n getComputedStyle(el).position === 'fixed' || getComputedStyle(el).position === 'sticky') {\n el.style.position = 'static';\n el.style.top = 'auto';\n el.style.zIndex = 'auto';\n }\n });\n }\n \n unstick();\n \n var observer = new MutationObserver(unstick);\n observer.observe(document.body, { childList: true, subtree: true, attributes: true, attributeFilter: ['style', 'class'] });\n})();", "Kill Sticky Headers");
}
} catch(__e) { console.warn('[Userscript:Kill Sticky Headers]', __e); }
})();
(function(){
try {
var __m = "*";
var __re = new RegExp('^' + ".*" + '

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An Inexpensive 'Homemade' O2/CO2 Regulating Incubator for Mammalian Cell Culture
Our incubator is an inexpensive, portable and user friendly device that is capable of regulating O2, CO2 and temperature within physiological ranges. Using an Arduino-based approach to control O2 and CO2 headspace levels via N2 and CO2 gas injection, we were able to convert a temperature-regulating Styrofoam egg incubator into a efficient benchtop cell culture tool comparable to commercial laboratory O2/CO2 incubators. This incubator can be assembled for less than $1000 (CAD), making it highly accessable for biologists seeking to transition towards physiological cell culture practices.
Table of Contents
General Information
  • Culturing cells under physiologically-relevant conditions is vital to maintaining cellular phenotypes representative of in vivo biology. While O2 constitutes ~20% of the atmosphere, it's level ranges from 2 to 6% in most mammalian tissues, thus O2/CO2 incubators capable of establishing such conditions are necessary. Unfortuanately, commercial O2/CO2 incubators suitable for mammalian cell culture cost well over CAD $10,000, making them inaccessible to many cell biologists.
  • The "Homemade" O2/CO2 incubator described here works to address this obstacle by providing cell culturists with the means to self assemble an effective O2/CO2 incubator from a standard temperature-regulating Styrofoam egg incubator.
  • The Stuart lab is a large proponent of the importance of physiological conditions in cell culture, thus by making tools to maintain physiological oxygen levels more readily available, we hope to encourage the field to incorporate such practices into standard workflow.
Project Road Map
  • 📖 [The Publication](Coming Soon) - This incubator was published in an international, peer-reviewed, open-access journal called Oxygen. All original designs and assembly details can be found in this manuscript + the supplementary data sections.
  • 💻 Arduino Code
  • 🛒 [Materials List] (Coming Soon)
  • ❓ Issue Forum - Submit questions, concerns and assembly requests here!
Technologies Used
Design
Initial PrototypePicture2
Final Model: gas sensors, arduino controller, soleniod valves and electronics enclosed within a 3D-printed boxPicture3
Setup
Usage
  • This benchtop incubator is primarily intended for academic research labs looking to incorporate physioxic cell culture conditions. The interior of the incubator is suitable for both adherent and suspension cell culture, comfortably accomodating at least 25 100mm stacked tissue culture disks and/or flasks.
  • Specific setpoint gas levels (O2 and CO2) can be altered to suit the conditions required by the user. The code is currently set to maintain a physioxic level of 5% O2, and standard 5% CO2.
Points to Keep in Mind
  • Depending on the application, the Uno and solenoid valves can be powered in one of two ways. For applications where the system is only run for short periods of time, both the Uno and solenoids can be powered with a single 12VDC power supply. However, if the application requires the Uno to be constantly powered, as in the incubator application developed here, the Uno should be powered with a lower voltage (9VDC), while the solenoid valves are powered independently with a 12VDC power supply.
  • Heatsinks may be required for various components including the transistors, though we did not find them necessary under our conditions. Monitoring the temperature of electrical components is critical in the early stages of development to determine the need for heatsinks.
Contact
Created by @StuartLab -- Feel free to contact us with questions or concerns about incubator assembly!
About
O2/CO2 incubators for cell culture
Resources
Readme
Activity
Stars
3 stars
Watchers
2 watching
Forks
1 fork
Report repository
Releases
Packages
Contributors

, 'i');
if (__m === '*' || __re.test(location.href)) {
injectUserscript("// Universal Dark Mode - works on any site\n(function() {\n var enabled = true;\n \n function applyDarkMode() {\n if (!enabled) return;\n \n // Create style element if it doesn't exist\n var style = document.getElementById('universal-dark-mode-style');\n if (!style) {\n style = document.createElement('style');\n style.id = 'universal-dark-mode-style';\n document.head.appendChild(style);\n }\n \n // Dark mode CSS - inverts colors but preserves images/video\n style.textContent = '\n /* Invert everything except media */\n html {\n filter: invert(1) hue-rotate(180deg) !important;\n background: #1a1a2e !important;\n }\n \n /* Restore images, videos, iframes, canvas */\n img, video, iframe, canvas, svg, picture, [style*=\"background-image\"] {\n filter: invert(1) hue-rotate(180deg) !important;\n }\n \n /* Preserve specific elements that should not be inverted */\n .no-dark-mode, .no-dark-mode *,\n [data-theme=\"light\"], [data-theme=\"light\"],\n .ace_editor, .ace_editor *,\n .CodeMirror, .CodeMirror *,\n .monaco-editor, .monaco-editor *,\n .markdown-body pre, .markdown-body pre *,\n .highlight, .highlight *,\n pre code, pre code * {\n filter: none !important;\n }\n \n /* Fix common UI elements */\n .modal, .popup, .dropdown-menu, .tooltip, .popover {\n filter: invert(1) hue-rotate(180deg) !important;\n background: #2d2d44 !important;\n border-color: #444 !important;\n }\n \n /* Scrollbars */\n ::-webkit-scrollbar { background: #1a1a2e !important; }\n ::-webkit-scrollbar-thumb { background: #444 !important; }\n ::-webkit-scrollbar-thumb:hover { background: #555 !important; }\n \n /* Selection */\n ::selection { background: #4ecdc4 !important; color: #1a1a2e !important; }\n ::-moz-selection { background: #4ecdc4 !important; color: #1a1a2e !important; }\n ';\n }\n \n function removeDarkMode() {\n var style = document.getElementById('universal-dark-mode-style');\n if (style) style.remove();\n }\n \n // Toggle with Alt+Shift+D\n document.addEventListener('keydown', function(e) {\n if (e.altKey && e.shiftKey && e.key === 'D') {\n e.preventDefault();\n enabled = !enabled;\n if (enabled) {\n applyDarkMode();\n console.log('[Universal Dark Mode] Enabled');\n } else {\n removeDarkMode();\n console.log('[Universal Dark Mode] Disabled');\n }\n }\n });\n \n // Apply on load\n applyDarkMode();\n \n // Re-apply on dynamic content\n var observer = new MutationObserver(function(mutations) {\n if (enabled && !document.getElementById('universal-dark-mode-style')) {\n applyDarkMode();\n }\n });\n observer.observe(document.head, { childList: true });\n \n console.log('[Universal Dark Mode] Loaded - Press Alt+Shift+D to toggle');\n})();", "Universal Dark Mode");
}
} catch(__e) { console.warn('[Userscript:Universal Dark Mode]', __e); }
})();
})();

Skip to content
BranchesTags
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History
10 Commits
Folders and files
NameName
Last commit message
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STL Files 3D Printable Parts
STL Files 3D Printable Parts
Arduino code
Arduino code
README.md
README.md
Repository files navigation
An Inexpensive 'Homemade' O2/CO2 Regulating Incubator for Mammalian Cell Culture
Our incubator is an inexpensive, portable and user friendly device that is capable of regulating O2, CO2 and temperature within physiological ranges. Using an Arduino-based approach to control O2 and CO2 headspace levels via N2 and CO2 gas injection, we were able to convert a temperature-regulating Styrofoam egg incubator into a efficient benchtop cell culture tool comparable to commercial laboratory O2/CO2 incubators. This incubator can be assembled for less than $1000 (CAD), making it highly accessable for biologists seeking to transition towards physiological cell culture practices.
Table of Contents
General Information
  • Culturing cells under physiologically-relevant conditions is vital to maintaining cellular phenotypes representative of in vivo biology. While O2 constitutes ~20% of the atmosphere, it's level ranges from 2 to 6% in most mammalian tissues, thus O2/CO2 incubators capable of establishing such conditions are necessary. Unfortuanately, commercial O2/CO2 incubators suitable for mammalian cell culture cost well over CAD $10,000, making them inaccessible to many cell biologists.
  • The "Homemade" O2/CO2 incubator described here works to address this obstacle by providing cell culturists with the means to self assemble an effective O2/CO2 incubator from a standard temperature-regulating Styrofoam egg incubator.
  • The Stuart lab is a large proponent of the importance of physiological conditions in cell culture, thus by making tools to maintain physiological oxygen levels more readily available, we hope to encourage the field to incorporate such practices into standard workflow.
Project Road Map
  • 📖 [The Publication](Coming Soon) - This incubator was published in an international, peer-reviewed, open-access journal called Oxygen. All original designs and assembly details can be found in this manuscript + the supplementary data sections.
  • 💻 Arduino Code
  • 🛒 [Materials List] (Coming Soon)
  • ❓ Issue Forum - Submit questions, concerns and assembly requests here!
Technologies Used
Design
Initial PrototypePicture2
Final Model: gas sensors, arduino controller, soleniod valves and electronics enclosed within a 3D-printed boxPicture3
Setup
Usage
  • This benchtop incubator is primarily intended for academic research labs looking to incorporate physioxic cell culture conditions. The interior of the incubator is suitable for both adherent and suspension cell culture, comfortably accomodating at least 25 100mm stacked tissue culture disks and/or flasks.
  • Specific setpoint gas levels (O2 and CO2) can be altered to suit the conditions required by the user. The code is currently set to maintain a physioxic level of 5% O2, and standard 5% CO2.
Points to Keep in Mind
  • Depending on the application, the Uno and solenoid valves can be powered in one of two ways. For applications where the system is only run for short periods of time, both the Uno and solenoids can be powered with a single 12VDC power supply. However, if the application requires the Uno to be constantly powered, as in the incubator application developed here, the Uno should be powered with a lower voltage (9VDC), while the solenoid valves are powered independently with a 12VDC power supply.
  • Heatsinks may be required for various components including the transistors, though we did not find them necessary under our conditions. Monitoring the temperature of electrical components is critical in the early stages of development to determine the need for heatsinks.
Contact
Created by @StuartLab -- Feel free to contact us with questions or concerns about incubator assembly!
About
O2/CO2 incubators for cell culture
Resources
Readme
Activity
Stars
3 stars
Watchers
2 watching
Forks
1 fork
Report repository
Releases
Packages
Contributors