Monodisperse droplet generator: On-chip Droplet Generator
On-chip Droplet Generator allows mass production of micrometer-sized water-in-oil (W/O) emulsion droplets and gel microdrops (GMDs) with a unique microfluidic chip. The individual droplets function as miniaturized vessels for reactions such as microbial/cell cultivation and genetic analysis. Droplet technology is now recognized as a prominent high-throughput screening method allowing single cell analysis or simultaneous assay of more than 1 million samples.
Exploration of novel microorganisms
Encapsulation, cultivation and isolation of a variety of microorganisms including soil microorganisms, intestinal microbiota and marine microorganisms.
Development of highly productive strains
Detection and separation of enzymes and metabolites secreted in droplets.
Screening of antibody-producing cells
Development of stable and productive antibody forming cell lines.
Generation of stable droplets
On-chip Droplet Generator allows production of stable and monodisperse droplets.
Easy adjustment of droplet size
Droplet size can be easily adjusted between 60 µm and 120 µm in diameter by regulating the pressure applied to the sample and oil reservoirs.
Real-time monitoring of droplet generation
Droplet size can be viewed and monitored real-time with the supplied monitoring device.
On-chip Droplet Generator can produce two types of droplets: water-in-oil (W/O) emulsion droplets and gel microdrops (GMDs).
Utilization of On-chip Droplet Generator and On-chip® Sort
Workflow of On-chip Droplet Generator and On-chip® Sort
Droplets produced by On-chip Droplet Generator can be screened at a high throughput manner by our microfluidic chip-based cell sorter, On-chip® Sort. On-chip® Sort is capable of using oil as the sheath fluid for analyzing and sorting of W/O emulsion droplets.
Sorting of droplets containing proliferated cells
W/O and GMD droplets containing proliferated cells encapsulated by On-chip Droplet Generator can be sorted on On-chip® Sort. Examples are:
(1) Sorting of E. coli encapsulated and incubated in W/O droplets based on the fluorescence of GFP (expression within E. coli as an indicator).
(2) Sorting of Tetrahymena encapsulated and incubated in W/O droplets based on the autofluorescence as an indicator.
(3) Sorting of A549 cells encapsulated and incubated in GMDs based on the fluorescence of GFP as an indicator.
Detection of microorganisms proliferated within W/O droplets
A simple and reliable method that allows bacteria containing droplets to be isolated on On-chip® Sort has been developed. W/O droplets containing bacteria were sorted based on the fluorescence produced as a result of the interaction between the bacteria and fluorescence resonance energy transfer (FRET)-based RNA probe encapsulated inside the droplets. This method is named Fluorescent Nucleic Acid Probe in droplets for Sorting bacteria (FNAP-sort). An increase in the fluorescence intensity of E. coli, Bacillus subtilis, Streptomyces aureofaciens or Bradyrhizobium japonicum cultivated as model microorganisms were identified in the droplets in which these microorganisms proliferated. The use of this system allows screening based on the proliferative activity in the environment and other cellular events.
Screening for high enzyme producing bacteria from soil
Microorganisms from the soil were encapsulated and incubated with the substrate which responded to enzyme (peptidase) in W/O droplets. One million droplets were analyzed and droplets with higher fluorescence intensity (i.e., high enzymatic activity) were sorted on On-chip® Sort. Collected droplets were transferred to agar plate. For those that formed colonies, supernatant assay was performed after re-incubation. Successful isolation of highly peptidase-producing microorganism was achieved using On-chip Droplet Generator, followed by On-chip® Sort.
Cultivation within W/O droplets maintains microbial diversity
Bacteria from soil were cultivated in liquid media and in W/O droplets over 7 days. DNA was extracted and analyzed by NGS to study the bacterial flora. The bacterial flora in the droplets showed to be much higher in individual Operational Taxonomic Units (OUTs) compared to those cultivated conventionally in liquid media.
Screening of high protein producer yeast with GMD
Yeast transformant that produces secreted luciferase were UV irradiated for random mutagenesis. They were then encapsulated and cultured within GMDs. GMDs with high fluorescence intensity derived from luciferase were sorted using On-chip® Sort and transferred to agar medium. For colonies formed, the supernatant was collected, and an assay was performed on the supernatant after re-incubation. As a result of screening, strains with higher producing ability than that of the parent strain were successfully collected.
Specifications of On-chip Droplet Generator
The On-chip Droplet Generator set up contains of the main unit for supplying pressure, monitoring unit for real-time monitoring of droplets, and temperature controller for precise temperature adjustment.
|Specifications||On-chip Droplet Generator
Monitoring unit with temperature control
|Main unit||Size||270 x 270 x 280 mm (W x D x H)|
|Liquid delivery pressure||7 – 80 kPa|
|Maximum liquid delivery line||3 lines (sample, oil, other reagents)|
|Droplet size generated||Diameter 60 – 120 μm (can be adjusted discretionally)|
|Generation rate||Approx. 100,000 droplets/3 min (for generation of 100 μm droplets)|
|Regulation||Laptop PC (Windows 10)|
|Power input||AC 100-240V, 50/60Hz|
|Power consumption||1.2A typ (ACIN 100V)|
|Monitoring unit||Size of monitoring unit||215 x 310 x 430 mm (W x D x H)|
|Accessories||Monitor camera, Peltier unit, Controller|
|Temperature range||-5℃ – 70℃|
|Ambient environment||Temperature 5 – 40˚C, humidity ≤ 85% RH|
|Power consumption||6.5A max|
|Product No.||Product name||Product description|
|60003||On-chip Droplet Generator
(Monitoring unit with temperature control)
|Main unit, chip holder, monitoring unit with temperature control|
|Samples applied||Aqueous solution, culture medium, gel solution etc.|
|Oils applied||Fluorinated oil|
|Fluid channel size||60 μm x 60 μm|
|Reservoir volume||Sample- 640 μL max.
Oil- 2,300 μL max.
|Output||Approx. 100,000 droplets (diameter 100 μm in approx. 3 min)|
|Product No.||Product name||Product description||Packing unit|
|63006||1060DG Chip Holder||Chip holder, tube rack, glass vials||1 piece|
|1003003||2D chip-1060DG||Droplet generation chip (10 chips/box)||10 chips/box|
Fluorinated oil is used for producing stable droplets.
|Product No.||Product name||Product description||Volume|
|2003003||On-chip® FluoroSurfactant-5wtH Oil||Fluorinated oil containing
|2003004||On-chip® FluoroSurfactant-0.1wtH Oil||Fluorinated oil containing