date: 2020-03-13T09:48:39Z pdf:PDFVersion: 1.5 pdf:docinfo:title: On-Chip Inverted Emulsion Method for Fast Giant Vesicle Production, Handling, and Analysis xmp:CreatorTool: LaTeX with hyperref package access_permission:can_print_degraded: true subject: Liposomes and giant unilamellar vesicles (GUVs) in particular are excellent compartments for constructing artificial cells. Traditionally, their use requires bench-top vesicle growth, followed by experimentation under a microscope. Such steps are time-consuming and can lead to loss of vesicles when they are transferred to an observation chamber. To overcome these issues, we present an integrated microfluidic chip which combines GUV formation, trapping, and multiple separate experiments in the same device. First, we optimized the buffer conditions to maximize both the yield and the subsequent trapping of the vesicles in micro-posts. Captured GUVs were monodisperse with specific size of 18 4 m in diameter. Next, we introduce a two-layer design with integrated valves which allows fast solution exchange in less than 20 s and on separate sub-populations of the trapped vesicles. We demonstrate that multiple experiments can be performed in a single chip with both membrane transport and permeabilization assays. In conclusion, we have developed a versatile all-in-one microfluidic chip with capabilities to produce and perform multiple experiments on a single batch of vesicles using low sample volumes. We expect this device will be highly advantageous for bottom-up synthetic biology where rapid encapsulation and visualization is required for enzymatic reactions. dc:format: application/pdf; version=1.5 pdf:docinfo:creator_tool: LaTeX with hyperref package access_permission:fill_in_form: true pdf:encrypted: false dc:title: On-Chip Inverted Emulsion Method for Fast Giant Vesicle Production, Handling, and Analysis modified: 2020-03-13T09:48:39Z cp:subject: Liposomes and giant unilamellar vesicles (GUVs) in particular are excellent compartments for constructing artificial cells. Traditionally, their use requires bench-top vesicle growth, followed by experimentation under a microscope. Such steps are time-consuming and can lead to loss of vesicles when they are transferred to an observation chamber. To overcome these issues, we present an integrated microfluidic chip which combines GUV formation, trapping, and multiple separate experiments in the same device. First, we optimized the buffer conditions to maximize both the yield and the subsequent trapping of the vesicles in micro-posts. Captured GUVs were monodisperse with specific size of 18 4 m in diameter. Next, we introduce a two-layer design with integrated valves which allows fast solution exchange in less than 20 s and on separate sub-populations of the trapped vesicles. We demonstrate that multiple experiments can be performed in a single chip with both membrane transport and permeabilization assays. In conclusion, we have developed a versatile all-in-one microfluidic chip with capabilities to produce and perform multiple experiments on a single batch of vesicles using low sample volumes. We expect this device will be highly advantageous for bottom-up synthetic biology where rapid encapsulation and visualization is required for enzymatic reactions. pdf:docinfo:subject: Liposomes and giant unilamellar vesicles (GUVs) in particular are excellent compartments for constructing artificial cells. Traditionally, their use requires bench-top vesicle growth, followed by experimentation under a microscope. Such steps are time-consuming and can lead to loss of vesicles when they are transferred to an observation chamber. To overcome these issues, we present an integrated microfluidic chip which combines GUV formation, trapping, and multiple separate experiments in the same device. First, we optimized the buffer conditions to maximize both the yield and the subsequent trapping of the vesicles in micro-posts. Captured GUVs were monodisperse with specific size of 18 4 m in diameter. Next, we introduce a two-layer design with integrated valves which allows fast solution exchange in less than 20 s and on separate sub-populations of the trapped vesicles. We demonstrate that multiple experiments can be performed in a single chip with both membrane transport and permeabilization assays. In conclusion, we have developed a versatile all-in-one microfluidic chip with capabilities to produce and perform multiple experiments on a single batch of vesicles using low sample volumes. We expect this device will be highly advantageous for bottom-up synthetic biology where rapid encapsulation and visualization is required for enzymatic reactions. pdf:docinfo:creator: Naresh Yandrapalli, Tina Seemann and Tom Robinson PTEX.Fullbanner: This is pdfTeX, Version 3.14159265-2.6-1.40.18 (TeX Live 2017/W32TeX) kpathsea version 6.2.3 meta:author: Naresh Yandrapalli trapped: False meta:creation-date: 2020-03-10T11:41:58Z created: 2020-03-10T11:41:58Z access_permission:extract_for_accessibility: true Creation-Date: 2020-03-10T11:41:58Z Author: Naresh Yandrapalli producer: pdfTeX-1.40.18 pdf:docinfo:producer: pdfTeX-1.40.18 pdf:unmappedUnicodeCharsPerPage: 17 dc:description: Liposomes and giant unilamellar vesicles (GUVs) in particular are excellent compartments for constructing artificial cells. Traditionally, their use requires bench-top vesicle growth, followed by experimentation under a microscope. Such steps are time-consuming and can lead to loss of vesicles when they are transferred to an observation chamber. To overcome these issues, we present an integrated microfluidic chip which combines GUV formation, trapping, and multiple separate experiments in the same device. First, we optimized the buffer conditions to maximize both the yield and the subsequent trapping of the vesicles in micro-posts. Captured GUVs were monodisperse with specific size of 18 4 m in diameter. Next, we introduce a two-layer design with integrated valves which allows fast solution exchange in less than 20 s and on separate sub-populations of the trapped vesicles. We demonstrate that multiple experiments can be performed in a single chip with both membrane transport and permeabilization assays. In conclusion, we have developed a versatile all-in-one microfluidic chip with capabilities to produce and perform multiple experiments on a single batch of vesicles using low sample volumes. We expect this device will be highly advantageous for bottom-up synthetic biology where rapid encapsulation and visualization is required for enzymatic reactions. Keywords: lab-on-chip; inverted emulsion method; microfluidics; giant vesicles; giant unilamellar vesicles (GUVs); bottom-up synthetic biology access_permission:modify_annotations: true dc:creator: Naresh Yandrapalli description: Liposomes and giant unilamellar vesicles (GUVs) in particular are excellent compartments for constructing artificial cells. Traditionally, their use requires bench-top vesicle growth, followed by experimentation under a microscope. Such steps are time-consuming and can lead to loss of vesicles when they are transferred to an observation chamber. To overcome these issues, we present an integrated microfluidic chip which combines GUV formation, trapping, and multiple separate experiments in the same device. First, we optimized the buffer conditions to maximize both the yield and the subsequent trapping of the vesicles in micro-posts. Captured GUVs were monodisperse with specific size of 18 4 m in diameter. Next, we introduce a two-layer design with integrated valves which allows fast solution exchange in less than 20 s and on separate sub-populations of the trapped vesicles. We demonstrate that multiple experiments can be performed in a single chip with both membrane transport and permeabilization assays. In conclusion, we have developed a versatile all-in-one microfluidic chip with capabilities to produce and perform multiple experiments on a single batch of vesicles using low sample volumes. We expect this device will be highly advantageous for bottom-up synthetic biology where rapid encapsulation and visualization is required for enzymatic reactions. dcterms:created: 2020-03-10T11:41:58Z Last-Modified: 2020-03-13T09:48:39Z dcterms:modified: 2020-03-13T09:48:39Z title: On-Chip Inverted Emulsion Method for Fast Giant Vesicle Production, Handling, and Analysis xmpMM:DocumentID: uuid:9dd58c45-c310-499a-bce7-33025fa0d4d4 Last-Save-Date: 2020-03-13T09:48:39Z pdf:docinfo:keywords: lab-on-chip; inverted emulsion method; microfluidics; giant vesicles; giant unilamellar vesicles (GUVs); bottom-up synthetic biology pdf:docinfo:modified: 2020-03-13T09:48:39Z meta:save-date: 2020-03-13T09:48:39Z pdf:docinfo:custom:PTEX.Fullbanner: This is pdfTeX, Version 3.14159265-2.6-1.40.18 (TeX Live 2017/W32TeX) kpathsea version 6.2.3 Content-Type: application/pdf X-Parsed-By: org.apache.tika.parser.DefaultParser creator: Naresh Yandrapalli dc:subject: lab-on-chip; inverted emulsion method; microfluidics; giant vesicles; giant unilamellar vesicles (GUVs); bottom-up synthetic biology access_permission:assemble_document: true xmpTPg:NPages: 11 pdf:charsPerPage: 3138 access_permission:extract_content: true access_permission:can_print: true pdf:docinfo:trapped: False meta:keyword: lab-on-chip; inverted emulsion method; microfluidics; giant vesicles; giant unilamellar vesicles (GUVs); bottom-up synthetic biology access_permission:can_modify: true pdf:docinfo:created: 2020-03-10T11:41:58Z