Hi Everyone!
I've been toying with some designs for a protoplast culture and PEG transformation chip and had a few questions:
1. Some of the only papers citing tobacco protoplast cultures and microfluidics have a design that consists of a wide channel which at one end have a sieve of posts a few microns in diameter and a few micros apart. This collects the cells, which range from 40-70um while replacing the media. This would cause an aggregate as the sieve fills and would damage cells...i think. Do you think an laser printer set to 1200 DPI can print a grid 20-30 microns apart? The post size won't matter as long as the sieving space is tight.
2. How is oxygenation and media recirculation done in bacterial culture chips? Do they just circulate the media without losing cells somehow?
3. Lets say I get protoplasts to make microcolonies, how (aside from cutting open the PDMS culture chamber) would I get them out to replate?
The whole idea of this chip is to repeatedly perform PEG fusions and naked plasmid uptake experiments on protoplasts and regenerate them. There is a lot of manual skill and tricks involved in PEG transformation so I want to take luck and dexterity out of the equation. Any advice would be fantastic.
4. Know any literature on standardized or good design practices for general microfluidics fabrication? I'm rockin PDMS-glass chips made using shrinky dinks and if it works Polyolefin shrink wrap chips too. I found a book that covers all the theory and fluid dynamics behind the curtain which is great but little on channel layout theory/advice/style.
5. Has anyone made valves using the shrinky dink method? The channels end up rounded which isn't the ideal geometry for the control layer.
Thanks in advance!
Sebastian S. Cocioba
CEO & Founder
New York Botanics, LLC
Plant Biotech R&D
I've been toying with some designs for a protoplast culture and PEG transformation chip and had a few questions:
1. Some of the only papers citing tobacco protoplast cultures and microfluidics have a design that consists of a wide channel which at one end have a sieve of posts a few microns in diameter and a few micros apart. This collects the cells, which range from 40-70um while replacing the media. This would cause an aggregate as the sieve fills and would damage cells...i think. Do you think an laser printer set to 1200 DPI can print a grid 20-30 microns apart? The post size won't matter as long as the sieving space is tight.
2. How is oxygenation and media recirculation done in bacterial culture chips? Do they just circulate the media without losing cells somehow?
3. Lets say I get protoplasts to make microcolonies, how (aside from cutting open the PDMS culture chamber) would I get them out to replate?
The whole idea of this chip is to repeatedly perform PEG fusions and naked plasmid uptake experiments on protoplasts and regenerate them. There is a lot of manual skill and tricks involved in PEG transformation so I want to take luck and dexterity out of the equation. Any advice would be fantastic.
4. Know any literature on standardized or good design practices for general microfluidics fabrication? I'm rockin PDMS-glass chips made using shrinky dinks and if it works Polyolefin shrink wrap chips too. I found a book that covers all the theory and fluid dynamics behind the curtain which is great but little on channel layout theory/advice/style.
5. Has anyone made valves using the shrinky dink method? The channels end up rounded which isn't the ideal geometry for the control layer.
Thanks in advance!
Sebastian S. Cocioba
CEO & Founder
New York Botanics, LLC
Plant Biotech R&D
0 comments:
Post a Comment