3D Printed Microscopy Chambers - Revision history

Admin: /* Chamber Assembly */

2023-02-23T16:59:50Z

Chamber Assembly

← Older revision		Revision as of 16:59, 23 February 2023
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	== Printing Chambers - FDM (Filament) Printing ==		== Printing Chambers - FDM (Filament) Printing ==
	FDM printing is the easiest in terms of printing and post-printing processing, ~~but~~ it is ~~more difficult~~ to ~~get a good seal with the coverslip than with DLP (resin) printing~~. It is important to use a good quality filament free of potentially cytotoxic or fluorescent compounds - we recommend a black or unpigmented, FDA-food grade certified PLA or PETG filament such as those sold by [https://filaments.ca/pages/search-results-page?q=fda Filaments.ca].		FDM printing is the easiest in terms of printing and post-printing processing, and it is easier to find biocomatable materials. It is important to use a good quality filament free of potentially cytotoxic or fluorescent compounds - we recommend a black or unpigmented, FDA-food grade certified PLA or PETG filament such as those sold by [https://filaments.ca/pages/search-results-page?q=fda Filaments.ca].
	[[File:Assembly.png\|thumb\|When printing, be certain that the coverslip-contacting part of the print faces upwards.]]		[[File:Assembly.png\|thumb\|When printing, be certain that the coverslip-contacting part of the print faces upwards.]]

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	* Ironing:		* Ironing:
	** Top-most surface for chambers (this helps with the bond to the coverslip)		** Top-most surface for chambers (this helps with the bond to the coverslip)
	** None on ~~Lids~~		** None on lids or alignment guides
	** All top surfaces for alignment guides
	* No supports		* No supports

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	== Printing Chambers - DLP (Resin) Printing ==		== Printing Chambers - DLP (Resin) Printing ==
	[[File:Resin.png\|thumb\|Only resin currently validated for cell-contact use.]]		[[File:Resin.png\|thumb\|Only resin currently validated for cell-contact use.]]
	DLP (resin) printing provides superior resolution and a flatter surface for bonding the coverslip to the chamber. However, the prints must be treated carefully after printing to remove any residual polymer or photocatalyst, as these are highly cytotoxic. At this time the only ~~verified~~ resin for ~~this purpose is~~ [https://www.amazon.ca/s?k=eSun+eResin-PLA+Bio-Photopolymer+Resin+Black&i=industrial&crid=L2CCXTG76DPP&sprefix=esun+eresin-pla+bio-photopolymer+resin+black%2Cindustrial%2C62&ref=nb_sb_noss eSun's eResin-PLA Bio-Photopolymer Resin (Black)].		DLP (resin) printing provides superior resolution and a flatter surface for bonding the coverslip to the chamber. However, the prints must be treated carefully after printing to remove any residual polymer or photocatalyst, as these are highly cytotoxic. At this time the only resin we've found that works for cell-contact purposes [https://www.amazon.ca/s?k=eSun+eResin-PLA+Bio-Photopolymer+Resin+Black&i=industrial&crid=L2CCXTG76DPP&sprefix=esun+eresin-pla+bio-photopolymer+resin+black%2Cindustrial%2C62&ref=nb_sb_noss eSun's eResin-PLA Bio-Photopolymer Resin (Black)].

	=== Recommended Print Settings ===		=== Recommended Print Settings ===
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	# Layout prints as desired in Lychee Slicer or Chitubox (or your preferred slicing software). Be certain to position the prints such that the surface that will contact the coverslip face upwards. The numbers and letters on the chamber will be upside-down if the chamber is positioned correctly.		# Layout prints as desired in Lychee Slicer or Chitubox (or your preferred slicing software). Be certain to position the prints such that the surface that will contact the coverslip face upwards. The numbers and letters on the chamber will be upside-down if the chamber is positioned correctly.
	# Rotate the prints 40° along the long axis (see image to right).		# Rotate the prints 40° along the long axis (see image to right).
	#Use autosupports to lift and generate supports for the chambers. A ~~minimum of~~ 3 mm lift is recommended.		#Use autosupports to lift and generate supports for the chambers. A 3 to 5 mm lift is recommended.
	#Manually remove any supports that contact the surface that will contact the coverslip as this surface needs to be absolutely smooth.		#Manually remove any supports that contact the surface that will contact the coverslip as this surface needs to be absolutely smooth.
	#Print		#Print
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	#Carefully remove the print from the print bed and place in a wash container of isopropyl alcohol. It is fine to use isopropyl that has been used previously for mashing prints in this step.		#Carefully remove the print from the print bed and place in a wash container of isopropyl alcohol. It is fine to use isopropyl that has been used previously for mashing prints in this step.
	#Using flat cutters carefully remove all supports.		#Using flat cutters carefully remove all supports.
	#Place the chambers in a ~~dedicated cleaning container, coverslip-contact side up~~. Do not stack the prints. Cover completely with clean (previously unused) isopropyl alcohol. ~~Wash with gentle shaking~~ for ~~20 minutes~~.		#Place the chambers in a ultrasonic cleaner. Do not stack the prints. Cover completely with clean (previously unused) isopropyl alcohol. Clean for 90 seconds.
	#Carefully remove the ~~chamber~~ and place in the curing station, coverslip-contact side up. Position the chambers such that they do not contact each other. Cure for 20 minutes, then flip the parts and cure an additional 20 minutes. Do not crowd parts during this step - it is better to cure in multiple batches than it is to have shadowed regions that do no cure.		#Carefully remove the chambers and rinse with cold tap water. Then place the prints in the curing station, coverslip-contact side up. Position the chambers such that they do not contact each other. Cure for 20 minutes, then flip the parts and cure an additional 20 minutes. Do not crowd parts during this step - it is better to cure in multiple batches than it is to have shadowed regions that do no cure.

	== Chamber Assembly ==		== Chamber Assembly ==
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	# Acid wash coverslips in advance of assembly day, ensuring they are completely dry and dust free before assembly.		# Acid wash coverslips in advance of assembly day, ensuring they are completely dry and dust free before assembly.
	# Place a coverslip into the bottom of the alignment guide.		# Place a coverslip into the bottom of the alignment guide.
	# ~~Place~~ a ~~small drop~~ of [https://siliconesolutions.com/ss-433t.html SS-433T silicone glue] (Silicone Solutions, OH, USA) on a ~~phenolic sheet~~, and using a ~~roller, spread it into a thin (~0.5 mm) thick layer that is larger than your chamber. Note that PDMS can be used as well (~~10:1 polymer:catalyst~~), but the bond will not be as strong~~.		# Apply the glue:
	~~# Press~~ the coverslip-~~contact part~~ of the chamber into the glue, and check for even glue across the entire sealing surface. If the whole surface is not coated, press the chamber into the glue again. Repeat until all coverslip-contact surfaces are whetted with glue.		## Option A: Paint a thin layer of [https://siliconesolutions.com/ss-433t.html SS-433T silicone glue] (Silicone Solutions, OH, USA) onto the coverslip-contacting surface of the chamber. This provides a stronger bond, but may be less biocompatable than PDMS.
			## Option B: Mix and degass PDMS, using a 10:1 polymer:catalyst ratio. Paint a thin layer onto the coverslip-contacting surface of the chamber.
	# Put the chamber (glue-side down) into the alignment guide and press onto the coverslip for 10 seconds.		# Put the chamber (glue-side down) into the alignment guide and press onto the coverslip for 10 seconds.
	# Carefully invert the alignment guide and remove the chamber.		# Carefully invert the alignment guide and remove the chamber.
	# Check the seal - a good seal will appear wet; any dry-appearing areas are not glued. If you see a dry area, gently press that part of the chamber with your finger. If a good seal cannot be made, remove the coverslip and repeat steps 2-6 with a new coverslip. This step is critical, as poorly glued chambers will leak.		# Check the seal - a good seal will appear wet; any dry-appearing areas are not glued. If you see a dry area, gently press that part of the chamber with your finger. If a good seal cannot be made, remove the coverslip and repeat steps 2-6 with a new coverslip. This step is critical, as poorly glued chambers will leak.
	# Transfer the completed chambers to a humidified, 37°C incubator for 24 hours to cure.		# Transfer the completed chambers to a humidified, 37°C incubator for 24 hours to cure.
			Optional: The strength of the PDMS bond can be increased by treating the chambers with an ~3 second exposure to oxygen plasma after step 7.

	== Chamber Sanitation ==		== Chamber Cleaning & Sanitation ==
	These chambers cannot be fully sterilized, but can be sanitized. As such, all cultures should be performed using media containing anitbiotic/anitmycotic solution.		These chambers cannot be fully sterilized, but can be sanitized. As such, all cultures should be performed using media containing anitbiotic/anitmycotic solution.

	# Soak the chambers, with gentle agitation, ~~in 70% ethanol or 70% isopropryl for 15 minutes.~~		# Soak the chambers, with gentle agitation, ddH<sub>2</sub>O, for 30 minutes. Replace the water and soak with agitation for an additional 30 minutes.
	~~# Rinse the chambers, , with gentle agitation, in autoclaved~~ ddH<sub>2</sub>O for 10 minutes. Dry ~~as much as possible~~.		# Dry using a slow flow of air from a compressed air valve.
	# Place into small zip-seal bags, with a matching lid, and seal.		# Place into small zip-seal bags, with a matching lid, and seal.
	# Place in the UV ~~chamber and irradiate for 30 minutes~~, being certain to not overlap/shadow any parts. ~~After 30 minutes~~, flip the ~~parts~~ and irradiate for ~~an additional 30 minutes~~.		# Place in the UV crosslinker, being certain to not overlap/shadow any parts. Irradiate for 5,000 mJ, then flip the chambers and irradiate for another 5,000 mJ.

	== 3D Printing Moulds for Casting PDMS Chambers ==		== 3D Printing Moulds for Casting PDMS Chambers ==

Admin at 18:31, 3 March 2022

2022-03-03T18:31:28Z

← Older revision		Revision as of 18:31, 3 March 2022
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	==Chamber Designs==		==Chamber Designs==
	Chambers are available to fit 18 mm x 18 mm and 24 mm x 50 mm coverslips, with different well arrangements. We are still testing some of these designs, but the images below ~~will link to~~ the ~~corresponding~~ .~~STL file once designs are finalized~~.		Chambers are available to fit 18 mm x 18 mm and 24 mm x 50 mm coverslips, with different well arrangements. We are still testing some of these designs, but the files can be downloaded at the following link. The images below show the file name, chamber layout, culture areas, and recommended media volumes for the varying chambers.

			* [http://phagocytes.ca/wp-content/uploads/2022/03/18mm.zip 18 mm chambers]
			* [http://phagocytes.ca/wp-content/uploads/2022/03/50mm.zip 24 x 50 mm chambers]

	{\| class="wikitable"		{\| class="wikitable"
	\|+
	![[File:50mm 1well.png\|thumb]]		![[File:50mm 1well.png\|thumb]]
	![[File:50mm 2well.png\|thumb]]		![[File:50mm 2well.png\|thumb]]
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	== Baseplates ==		== Baseplates ==
	These baseplates are designed to hold the 18 mm or 50 mm chambers in a Chamlide TC Live Cell Chamber. These should be printed in black PETG for temperature resistance and durability, 0.2 mm layer height. The bottom plates require supports, and it helps to iron (smooth) all top surfaces. The small circular inserts hold 4 mm diameter, 1 mm thick, disk magnets. These hold the top-plates in-place during imaging. To ensure the lid and base magnets lineup properly, a ~~magnetic~~ alignment tool is provided. Glue 4 mm diameter x 1 mm thick disk magnets onto either end of the alignment tool, ensuring that the pole of the magnet on the 'B' end is the opposite pole of the magnet on the 'L' end (e.g. North for 'B', South for 'L'). Use the tool to hold onto, and place, magnets into the lid ('L') and Base/Bottom ('B'). Place a small drop of instant (cyanoacrylate) glue into each circular insert, and then use the 'L' magnet to hold and place the magnets for the lids, and the 'B' magnet to hold and place the magnets for the base. This will ensure that the magnets in the base will attract the magnets in the lid, holding the chamber together. ~~We are still testing some of these designs, but~~ the ~~images below will link to the corresponding~~ .~~STL file once designs are finalized~~.		These baseplates are designed to hold the 18 mm or 50 mm chambers in a Chamlide TC Live Cell Chamber. These should be printed in black PETG for temperature resistance and durability, 0.2 mm layer height. The bottom plates require supports, and it helps to iron (smooth) all top surfaces. The small circular inserts hold 4 mm diameter, 1 mm thick, disk magnets. These hold the top-plates in-place during imaging. To ensure the lid and base magnets lineup properly, a magnet alignment tool is provided. Glue 4 mm diameter x 1 mm thick disk magnets onto either end of the alignment tool, ensuring that the pole of the magnet on the 'B' end is the opposite pole of the magnet on the 'L' end (e.g. North for 'B', South for 'L'). Use the tool to hold onto, and place, magnets into the lid ('L') and Base/Bottom ('B'). Place a small drop of instant (cyanoacrylate) glue into each circular insert, and then use the 'L' magnet to hold and place the magnets for the lids, and the 'B' magnet to hold and place the magnets for the base. This will ensure that the magnets in the base will attract the magnets in the lid, holding the chamber together. The STL files for both stages and the magnet alignment tool can be [http://phagocytes.ca/wp-content/uploads/2022/03/Base-Plates.zip downloaded here].
	{\|		{\|
	![[File:18mm BasePlate.png\|thumb]]		![[File:18mm BasePlate.png\|thumb]]

Admin at 16:26, 3 March 2022

2022-03-03T16:26:32Z

← Older revision		Revision as of 16:26, 3 March 2022
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	Customized imaging chambers can be 3D printed using FDM or SLA printing technology, and attached to a #1.5 thickness coverglass to allow for high-resolution imaging. When prepared properly, these chambers show no cytotoxicity or other signs of altered cell behaviour, even in cells cultured for multiple days. These ~~are the~~ chamber designs used in the [https://www.phagocytes.ca Heit Lab], and are ~~based on~~ our publication in Biochemistry and Cell Biology<ref name=":0">Tepperman A, Zheng DJ, Taka MA, Vrieze A, Le Lam A, Heit B. '''Customizable live-cell imaging chambers for multimodal and multiplex fluorescence microscopy.''' ''Biochem Cell Biol''. 2020 Oct;98(5):612-623. doi: 10.1139/bcb-2020-0064. Epub 2020 Apr 27. PMID: [https://pubmed.ncbi.nlm.nih.gov/32339465/ 32339465]. [https://cdnsciencepub.com/doi/10.1139/bcb-2020-0064 Link to Paper]. [https://www.biorxiv.org/content/10.1101/2020.02.19.955971v1 Link to Preprint].</ref>, ~~and~~ on the Mod3D preprint<ref>C. Barba Bazan, S. Goss, C. Peng, N. Begeja, CE. Suart, K. Neuman, Ray Truant. '''Mod3D: A Low-Cost, Flexible Modular System of Live-Cell Microscopy Chambers and Holders.''' ''bioRxiv'' 2021.10.18.462400; doi: https://doi.org/10.1101/2021.10.18.462400</ref> by the [https://raytruantlab.ca/ Truant Lab].		Customized imaging chambers can be 3D printed using FDM or SLA printing technology, and attached to a #1.5 thickness coverglass to allow for high-resolution imaging. When prepared properly, these chambers show no cytotoxicity or other signs of altered cell behaviour, even in cells cultured for multiple days. These chamber designs used in the [https://www.phagocytes.ca Heit Lab], and are the newest version of the chambers presented in our publication in Biochemistry and Cell Biology<ref name=":0">Tepperman A, Zheng DJ, Taka MA, Vrieze A, Le Lam A, Heit B. '''Customizable live-cell imaging chambers for multimodal and multiplex fluorescence microscopy.''' ''Biochem Cell Biol''. 2020 Oct;98(5):612-623. doi: 10.1139/bcb-2020-0064. Epub 2020 Apr 27. PMID: [https://pubmed.ncbi.nlm.nih.gov/32339465/ 32339465]. [https://cdnsciencepub.com/doi/10.1139/bcb-2020-0064 Link to Paper]. [https://www.biorxiv.org/content/10.1101/2020.02.19.955971v1 Link to Preprint].</ref>, modified based on some exciting advances in the Mod3D preprint<ref>C. Barba Bazan, S. Goss, C. Peng, N. Begeja, CE. Suart, K. Neuman, Ray Truant. '''Mod3D: A Low-Cost, Flexible Modular System of Live-Cell Microscopy Chambers and Holders.''' ''bioRxiv'' 2021.10.18.462400; doi: https://doi.org/10.1101/2021.10.18.462400</ref> by the [https://raytruantlab.ca/ Truant Lab].

	In addition to directly printing chambers, 3D printing can also be used to make moulds for casting PDMS chambers, as described in our publication in Biochemistry and Cell Biology<ref name=":0" />.		In addition to directly printing chambers, 3D printing can also be used to make moulds for casting PDMS chambers, as described in our publication in Biochemistry and Cell Biology<ref name=":0" />.

Admin: /* DLP Printing */

2022-03-03T15:44:07Z

DLP Printing

← Older revision		Revision as of 15:44, 3 March 2022
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	# Rotate the prints 40° along the long axis (see image to right).		# Rotate the prints 40° along the long axis (see image to right).
	#Use autosupports to lift and generate supports for the chambers. A minimum of 3 mm lift is recommended.		#Use autosupports to lift and generate supports for the chambers. A minimum of 3 mm lift is recommended.
			#Manually remove any supports that contact the surface that will contact the coverslip as this surface needs to be absolutely smooth.
	#Print		#Print
	#The prints are soft at this point, so treat them gently for the next few steps.		#The prints are soft at this point, so treat them gently for the next few steps.
	#Carefully remove the print from the print bed and place in a wash container of isopropyl alcohol. It is fine to use isopropyl that has been used previously for this step.		#Carefully remove the print from the print bed and place in a wash container of isopropyl alcohol. It is fine to use isopropyl that has been used previously for mashing prints in this step.
	#Using flat cutters carefully remove all supports.		#Using flat cutters carefully remove all supports.
	#Place the chambers in a dedicated cleaning container, coverslip-contact side up. Do not stack the prints. Cover completely with clean (previously unused) isopropyl alcohol. Wash with gentle shaking for 20 minutes.		#Place the chambers in a dedicated cleaning container, coverslip-contact side up. Do not stack the prints. Cover completely with clean (previously unused) isopropyl alcohol. Wash with gentle shaking for 20 minutes.

Admin: /* FDM (Filament) Printing */

2022-03-03T15:42:02Z

FDM (Filament) Printing

← Older revision		Revision as of 15:42, 3 March 2022
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	\|}		\|}

	== FDM (Filament) Printing ==		== Printing Chambers - FDM (Filament) Printing ==
	FDM printing is the easiest in terms of printing and post-printing processing, but it is more difficult to get a good seal with the coverslip than with DLP (resin) printing. It is important to use a good quality filament free of potentially cytotoxic or fluorescent compounds - we recommend a black or unpigmented, FDA-food grade certified PLA or PETG filament such as those sold by [https://filaments.ca/pages/search-results-page?q=fda Filaments.ca].		FDM printing is the easiest in terms of printing and post-printing processing, but it is more difficult to get a good seal with the coverslip than with DLP (resin) printing. It is important to use a good quality filament free of potentially cytotoxic or fluorescent compounds - we recommend a black or unpigmented, FDA-food grade certified PLA or PETG filament such as those sold by [https://filaments.ca/pages/search-results-page?q=fda Filaments.ca].
	[[File:Assembly.png\|thumb\|When printing, be certain that the coverslip-contacting part of the print faces upwards.]]		[[File:Assembly.png\|thumb\|When printing, be certain that the coverslip-contacting part of the print faces upwards.]]
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	# Dry chamber before assembly (see below).		# Dry chamber before assembly (see below).

	== DLP (Resin) Printing ==		== Printing Chambers - DLP (Resin) Printing ==
	[[File:Resin.png\|thumb\|Only resin currently validated for cell-contact use.]]		[[File:Resin.png\|thumb\|Only resin currently validated for cell-contact use.]]
	DLP (resin) printing provides superior resolution and a flatter surface for bonding the coverslip to the chamber. However, the prints must be treated carefully after printing to remove any residual polymer or photocatalyst, as these are highly cytotoxic. At this time the only verified resin for this purpose is [https://www.amazon.ca/s?k=eSun+eResin-PLA+Bio-Photopolymer+Resin+Black&i=industrial&crid=L2CCXTG76DPP&sprefix=esun+eresin-pla+bio-photopolymer+resin+black%2Cindustrial%2C62&ref=nb_sb_noss eSun's eResin-PLA Bio-Photopolymer Resin (Black)].		DLP (resin) printing provides superior resolution and a flatter surface for bonding the coverslip to the chamber. However, the prints must be treated carefully after printing to remove any residual polymer or photocatalyst, as these are highly cytotoxic. At this time the only verified resin for this purpose is [https://www.amazon.ca/s?k=eSun+eResin-PLA+Bio-Photopolymer+Resin+Black&i=industrial&crid=L2CCXTG76DPP&sprefix=esun+eresin-pla+bio-photopolymer+resin+black%2Cindustrial%2C62&ref=nb_sb_noss eSun's eResin-PLA Bio-Photopolymer Resin (Black)].
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	#Carefully remove the chamber and place in the curing station, coverslip-contact side up. Position the chambers such that they do not contact each other. Cure for 20 minutes, then flip the parts and cure an additional 20 minutes. Do not crowd parts during this step - it is better to cure in multiple batches than it is to have shadowed regions that do no cure.		#Carefully remove the chamber and place in the curing station, coverslip-contact side up. Position the chambers such that they do not contact each other. Cure for 20 minutes, then flip the parts and cure an additional 20 minutes. Do not crowd parts during this step - it is better to cure in multiple batches than it is to have shadowed regions that do no cure.

	== Assembly ==		== Chamber Assembly ==
	Assembly is the same whether the chambers are printed via DLP or FDM.		Assembly is the same whether the chambers are printed via DLP or FDM.

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	# Transfer the completed chambers to a humidified, 37°C incubator for 24 hours to cure.		# Transfer the completed chambers to a humidified, 37°C incubator for 24 hours to cure.

	== Sanitation ==		== Chamber Sanitation ==
	These chambers cannot be fully sterilized, but can be sanitized. As such, all cultures should be performed using media containing anitbiotic/anitmycotic solution.		These chambers cannot be fully sterilized, but can be sanitized. As such, all cultures should be performed using media containing anitbiotic/anitmycotic solution.

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	# Place in the UV chamber and irradiate for 30 minutes, being certain to not overlap/shadow any parts. After 30 minutes, flip the parts and irradiate for an additional 30 minutes.		# Place in the UV chamber and irradiate for 30 minutes, being certain to not overlap/shadow any parts. After 30 minutes, flip the parts and irradiate for an additional 30 minutes.

	== PDMS ~~Moulds~~ ==		== 3D Printing Moulds for Casting PDMS Chambers ==
	Both FDM and DLP can be used to prepare PDMS moulds, which can be used to cast PDMS chambers. FDM requires the minimal preparation, but has lower resolution than DLP.		Both FDM and DLP can be used to prepare PDMS moulds, which can be used to cast PDMS chambers. FDM requires the minimal preparation, but has lower resolution than DLP.

Admin at 15:39, 3 March 2022

2022-03-03T15:39:28Z

@@ Line 38: / Line 38: @@
 |[[File:18mm Lid Clear.png|thumb]]
+|
 |}

Admin: /* DLP Printing */

2022-03-02T19:03:22Z

DLP Printing

← Older revision		Revision as of 19:03, 2 March 2022
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	#Carefully remove the print from the print bed and place in a wash container of isopropyl alcohol. It is fine to use isopropyl that has been used previously for this step.		#Carefully remove the print from the print bed and place in a wash container of isopropyl alcohol. It is fine to use isopropyl that has been used previously for this step.
	#Using flat cutters carefully remove all supports.		#Using flat cutters carefully remove all supports.
	#Place the chambers in a dedicated cleaning container, coverslip-contact side up. Do not stack the prints. Cover completely with clean (previously unused) isopropyl alcohol. Wash with ~~gently~~ shaking for 20 minutes.		#Place the chambers in a dedicated cleaning container, coverslip-contact side up. Do not stack the prints. Cover completely with clean (previously unused) isopropyl alcohol. Wash with gentle shaking for 20 minutes.
	#Carefully remove the chamber and place in the curing station, coverslip-contact side up. Position the chambers such that they do not contact each other. Cure for 20 minutes, then flip the parts and cure an additional 20 minutes. Do not crowd parts during this step - it is better to cure in multiple batches than it is to have shadowed regions that do no cure.		#Carefully remove the chamber and place in the curing station, coverslip-contact side up. Position the chambers such that they do not contact each other. Cure for 20 minutes, then flip the parts and cure an additional 20 minutes. Do not crowd parts during this step - it is better to cure in multiple batches than it is to have shadowed regions that do no cure.

Admin: /* DLP Printing */

2022-03-02T19:02:41Z

DLP Printing

← Older revision		Revision as of 19:02, 2 March 2022
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	# Layout prints as desired in Lychee Slicer or Chitubox (or your preferred slicing software). Be certain to position the prints such that the surface that will contact the coverslip face upwards. The numbers and letters on the chamber will be upside-down if the chamber is positioned correctly.		# Layout prints as desired in Lychee Slicer or Chitubox (or your preferred slicing software). Be certain to position the prints such that the surface that will contact the coverslip face upwards. The numbers and letters on the chamber will be upside-down if the chamber is positioned correctly.
	# Rotate the prints 40° ~~on the short axis (e.g. objects are rotated~~ along the long axis, image to right).		# Rotate the prints 40° along the long axis (see image to right).
	#Use autosupports to lift and generate supports for the chambers. A minimum of 3 mm lift is recommended.		#Use autosupports to lift and generate supports for the chambers. A minimum of 3 mm lift is recommended.
	#Print		#Print

Admin: /* FDM (Filament) Printing */

2022-03-02T19:01:53Z

FDM (Filament) Printing

← Older revision		Revision as of 19:01, 2 March 2022
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	== FDM (Filament) Printing ==		== FDM (Filament) Printing ==
	FDM printing is the easiest in terms of printing and post-printing ~~assembly~~, but it is more difficult to get a good seal with the coverslip than with DLP (resin) printing. It is important to use a good quality filament free of potentially cytotoxic or fluorescent compounds - we recommend a black or unpigmented, FDA-food grade certified PLA or PETG filament such as those sold by [https://filaments.ca/pages/search-results-page?q=fda Filaments.ca].		FDM printing is the easiest in terms of printing and post-printing processing, but it is more difficult to get a good seal with the coverslip than with DLP (resin) printing. It is important to use a good quality filament free of potentially cytotoxic or fluorescent compounds - we recommend a black or unpigmented, FDA-food grade certified PLA or PETG filament such as those sold by [https://filaments.ca/pages/search-results-page?q=fda Filaments.ca].
	[[File:Assembly.png\|thumb\|When printing, be certain that the coverslip-contacting part of the print faces upwards.]]		[[File:Assembly.png\|thumb\|When printing, be certain that the coverslip-contacting part of the print faces upwards.]]

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	== DLP (Resin) Printing ==		== DLP (Resin) Printing ==
	[[File:Resin.png\|thumb\|Only resin currently validated for cell-contact use.]]		[[File:Resin.png\|thumb\|Only resin currently validated for cell-contact use.]]
	DLP (resin) printing provides superior resolution and a flatter surface for bonding the coverslip to the chamber. However, the prints must be treated carefully to remove any residual polymer or photocatalyst, as these are highly cytotoxic. At this time the only verified resin for this purpose is [https://www.amazon.ca/s?k=eSun+eResin-PLA+Bio-Photopolymer+Resin+Black&i=industrial&crid=L2CCXTG76DPP&sprefix=esun+eresin-pla+bio-photopolymer+resin+black%2Cindustrial%2C62&ref=nb_sb_noss eSun's eResin-PLA Bio-Photopolymer Resin (Black)].		DLP (resin) printing provides superior resolution and a flatter surface for bonding the coverslip to the chamber. However, the prints must be treated carefully after printing to remove any residual polymer or photocatalyst, as these are highly cytotoxic. At this time the only verified resin for this purpose is [https://www.amazon.ca/s?k=eSun+eResin-PLA+Bio-Photopolymer+Resin+Black&i=industrial&crid=L2CCXTG76DPP&sprefix=esun+eresin-pla+bio-photopolymer+resin+black%2Cindustrial%2C62&ref=nb_sb_noss eSun's eResin-PLA Bio-Photopolymer Resin (Black)].

	=== Recommended Print Settings ===		=== Recommended Print Settings ===

Admin at 19:01, 2 March 2022

2022-03-02T19:01:16Z

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