A Cascade Microfluidic Device for High Quality Mitochondria Extraction

2015 ◽  
Author(s):  
Ching-Wen Li ◽  
Pao-Hsin Yen ◽  
Gou-Jen Wang
Keyword(s):  
Microfluidic Device ◽  
Complex I ◽  
Soft Lithography ◽  
Cell Damage ◽  
High Quality ◽  
Protein Assay ◽  
Lithography Process ◽  
Disease Therapy ◽  
Sds Page ◽  
Total Protein Assay

Recent researches have demonstrated that cells ingest mitochondrial by endocytosis to repair cell damage. This new mitochondrial therapy approach can be used for curing particular disease of neuropathy related diseases. Hence the obtainment of high quality and healthy mitochondria play an important role in mitochondrial based disease therapy. In this study, we propose a cascade microfluidic device for green extraction of healthy mitochondria. The geometry of the device was designed using the commercially available COMSOL package. Soft lithography process was than conducted to realize the device of PDMS. We used C2-GFP cells to demonstrate the efficiency of the proposed cascade microfluidic device. The total protein assay kit (complex I-V) was conducted to examine the extractive protein and the SDS page (Tom 20) was used for measuring the activity of the extracted mitochondria. Experimental results illustrate that the complex I-V expression of the extracted mitochondria by the proposed device is much higher than that of the extracted mitochondria by conventional kit. Furthermore, the results of the Tom 20 expression also demonstrate that our device is able to extract more amounts of mitochondria than the conventional kit.

Prediction of Human Acute Toxicity by the Hep G2/24-hour/Total Protein Assay, with Protein Measurement by the CBQCA Method

2005 ◽  
Vol 33 (3) ◽  
pp. 207-213 ◽  
Author(s):  
Paul J. Dierickx
Keyword(s):  
Protein Content ◽  
Total Protein ◽  
Total Protein Content ◽  
Human Toxicity ◽  
Hep G2 ◽  
Vitro Assay ◽  
Protein Assay ◽  
Lowry Method ◽  
Total Protein Assay

In our previously described Hep G2/24-hour/total protein assay, protein levels were measured by using the Lowry method. This assay was the best acute in vitro assay for the prediction of human toxicity within the Multicentre Evaluation of In Vitro Cytotoxicity (MEIC) study. In order to increase the MEIC data-base with a wider range of chemicals, we were interested in introducing the more practical 3-(4-carboxybenzoyl)-quinoline-2-carboxaldehyde (CBQCA) method for the quantification of the total protein content. Therefore, we investigated whether the same good results for the prediction of acute human toxicity would be obtained with the CBQCA method. The cells were treated for 24 hours, then cytotoxicity was determined by measuring the total protein content with CBQCA. The results were quantified by using the PI50c: the concentration (in mM) of test compound required to reduce the total protein content measured with the CBQCA-method by 50% as compared to the control cells. The results were compared with the PI50, the corresponding value when the Lowry method was used. A relatively low correlation was observed between PI50 and PI50c, reflecting the large and unexpected, differences when using the two protein assays. However, when comparing the log PI50c with the human toxicity, a correlation coefficient of r2 = 0.761 ( n = 44) was obtained for exactly the same series of MEIC chemicals. This value is clearly higher than that for the Lowry method ( r2 = 0.695). Compared to the Lowry method originally used, the Hep G2/24-hour/CBQCA total protein assay has the additional important advantage that it can be very easily adapted for large-scale analyses with robotic systems, including the on-line calculation of the results.

Erratum — Universal method for fabricating PDMS microfluidic device using SU8, 3D printing and soft lithography

2021 ◽  
pp. 1-1
Author(s):  
Charmi Chande ◽  
Nida Riaz ◽  
Andrew House ◽  
Victoria Harbour ◽  
Hathija Noor ◽  
...  
Keyword(s):  
3D Printing ◽  
Microfluidic Device ◽  
Soft Lithography ◽  
Universal Method

High Quality Epitaxial Growth on in-Situ Patterned Inp Substrates

1989 ◽  
Vol 145 ◽  
Author(s):  
H. Temkin ◽  
L. R. Harriott ◽  
J. Weiner ◽  
R. A. Hamm ◽  
M. B. Panish
Keyword(s):  
Epitaxial Growth ◽  
Ion Beam ◽  
Low Energy ◽  
High Quality ◽  
Lithography Process ◽  
Luminescence Efficiency ◽  
Surface Steps ◽  
Inp Substrates ◽  
Vacuum Lithography

AbstractWe demonstrate a vacuum lithography process which uses a finely focused Ga ion beam to write the pattern which is then transferred to the InP pattern by low energy dry etching. Surface steps on the order of 1000-2000A in height, and lateral resolution limited only by size of the ion beam, can be efficiently prepared using moderate Ga ion fluences. The surfaces prepared by this process are damage free and suitable for epitaxial overgrowth. GaInAs/InP heterostructures grown on in-situ patterned substrates show excellent morphology and high luminescence efficiency.

scholarly journals Micropipette-Based Microfluidic Device for Monodisperse Microbubbles Generation

Micromachines ◽  
2018 ◽  
Vol 9 (8) ◽  
pp. 387
Author(s):  
Carlos Toshiyuki Matsumi ◽  
Wilson José da Silva ◽  
Fábio Kurt Schneider ◽  
Joaquim Miguel Maia ◽  
Rigoberto E. M. Morales ◽  
...  
Keyword(s):  
Electric Fields ◽  
Microfluidic Device ◽  
Soft Lithography ◽  
Low Cost ◽  
Characteristic Curve ◽  
Microfluidic Devices ◽  
Average Diameter ◽  
Internal Diameter ◽  
Medical Diagnostic ◽  
Average Size

Microbubbles have various applications including their use as carrier agents for localized delivery of genes and drugs and in medical diagnostic imagery. Various techniques are used for the production of monodisperse microbubbles including the Gyratory, the coaxial electro-hydrodynamic atomization (CEHDA), the sonication methods, and the use of microfluidic devices. Some of these techniques require safety procedures during the application of intense electric fields (e.g., CEHDA) or soft lithography equipment for the production of microfluidic devices. This study presents a hybrid manufacturing process using micropipettes and 3D printing for the construction of a T-Junction microfluidic device resulting in simple and low cost generation of monodisperse microbubbles. In this work, microbubbles with an average size of 16.6 to 57.7 μm and a polydispersity index (PDI) between 0.47% and 1.06% were generated. When the device is used at higher bubble production rate, the average diameter was 42.8 μm with increased PDI of 3.13%. In addition, a second-order polynomial characteristic curve useful to estimate micropipette internal diameter necessary to generate a desired microbubble size is presented and a linear relationship between the ratio of gaseous and liquid phases flows and the ratio of microbubble and micropipette diameters (i.e., Qg/Ql and Db/Dp) was found.

Highly stretchable conductive polypyrrole film on a three dimensional porous polydimethylsiloxane surface fabricated by a simple soft lithography process

RSC Advances ◽  
2016 ◽  
Vol 6 (114) ◽  
pp. 113344-113351 ◽  
Author(s):  
Chia-Wen Tsao ◽  
Xu-Cheng Guo ◽  
Wei-Wen Hu
Keyword(s):  
Soft Lithography ◽  
Low Cost ◽  
Three Dimensional ◽  
Lithography Process ◽  
Conductive Substrate ◽  
Polypyrrole Film ◽  
Highly Stretchable ◽  
Porous Microstructures ◽  
Abrasive Paper ◽  
Conductive Polypyrrole

We create an elastic porous polydimethylsiloxane highly stretchable conductive substrate. The surface is fabricated by a simple soft lithography process that replicates the 3D corrugated porous microstructures from a low-cost commercially available abrasive paper.

scholarly journals Effects of Fluoride on the Interactions between Amelogenin and Apatite Crystals

2008 ◽  
Vol 87 (1) ◽  
pp. 39-44 ◽  
Author(s):  
K. Tanimoto ◽  
T. Le ◽  
L. Zhu ◽  
J. Chen ◽  
J.D.B. Featherstone ◽  
...  
Keyword(s):  
Crystal Growth ◽  
Crystal Surface ◽  
Titration Calorimetry ◽  
Carbonated Hydroxyapatite ◽  
Protein Assay ◽  
Binding Rate ◽  
Apatite Crystals ◽  
Sds Page ◽  
Initial Binding

Fluorosed enamel is more porous and less mineralized, possibly related to altered amelogenin-modulated crystal growth. The purpose of this study was to examine the role of fluoride in interactions between amelogenin and apatite crystals. Recombinant human amelogenin (rh174) was bound to carbonated hydroxyapatite containing various amounts of fluoride, and analyzed by protein assay, SDS PAGE, and AFM. Interactions between rh174 and fluoride were assayed by isothermal titration calorimetry (ITC). The initial binding rate of rh174, as well as total amount of rh174 bound to fluoride-containing carbonated hydroxyapatite, was greater than that in the control carbonated hydroxyapatite. Fluoride in solution at physiologic (5.3 micromolar, or 0.1 ppm) concentrations showed no significant effect on binding, but higher fluoride levels significantly decreased protein binding. ITC showed no interactions between fluoride and rh174. These results suggest that fluoride incorporation into the crystal lattice alters the crystal surface to enhance amelogenin binding, with no direct interactions between fluoride and amelogenin.

The Electrophoretic Contribution to the Solutions of Taxonomical Problems of Some Lathyrus L. Taxa Growing in Turkey

2020 ◽  
Author(s):  
H. Genc ◽  
İ. Emre ◽  
A. Sahin
Keyword(s):  
Seed Storage Protein ◽  
Extraction Procedure ◽  
Seed Proteins ◽  
Seed Storage ◽  
Morphological Data ◽  
Protein Patterns ◽  
Documentation System ◽  
Natural Habitats ◽  
Protein Assay ◽  
Sds Page

Background: The seed proteins are used as molecular markers to clarify the systematic problems. Also, electrophoretic techniques are safe tools to identify the seed proteins. In present study, it was used the SDS-PAGE technique to solve the taxonomical problems of eight taxa of genus Lathyrus belong to three sections Orobus, Lathyrostylis and Pratensis according to the globulin B and glutelin. Methods: The seed materials were collected from natural habitats and 0.5 g seed were homogenized and centrifuged based on extraction procedure. In addition, the amounts of seed storage protein subfractions were determined by using protein assay. The electrophoretic analysis were performed according to the 12% SDS-PAGE. The gel documentation system (Bio-Rad, USA) was used to analyse the electrophoretic data and UPGAMA was used to construct the dendogram to show the relationships among the species under focus. Results: Current study showed that the species of section Pratensis were different from species of sections Orobus and Lathyrostylis. The differences among the studied taxa shown clearly and all studied taxa were choosen from the protein patterns. Furthermore, present study demonstrated that L. nivalis has the highest globulin B and glutelin. Results of the present study generally supported the morphological data.

Sign in / Sign up

Export Citation Format

Share Document