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Enzyme Kinetics01:19

Enzyme Kinetics

104.1K
Enzymes speed up reactions by lowering the activation energy of the reactants. The speed at which the enzyme turns reactants into products is called the rate of reaction. Several factors impact the rate of reaction, including the number of available reactants. Enzyme kinetics is the study of how an enzyme changes the rate of a reaction.
Scientists typically study enzyme kinetics with a fixed amount of enzyme in the controlled environment of a test tube. When more reactant, or substrate, is...
104.1K
Introduction to Enzyme Kinetics01:19

Introduction to Enzyme Kinetics

33.7K
Enzyme kinetics studies the rates of biochemical reactions. Scientists monitor the reaction rates for a particular enzymatic reaction at various substrate concentrations. Additional trials with inhibitors or other molecules that affect the reaction rate may also be performed.
The experimenter can then plot the initial reaction rate or velocity (Vo) of a given trial against the substrate concentration ([S]) to obtain a graph of the reaction properties. For many enzymatic reactions involving a...
33.7K
Mass Spectrometry: Overview01:19

Mass Spectrometry: Overview

8.9K
Mass spectrometry is an analytical technique used to determine the molecular mass and molecular formula of a compound. The basic principle of mass spectrometry is to generate ions from the analyte molecule and measure these ion abundances against their molecular mass. One common type of ionization, known as electron ionization or EI, bombards the analyte molecules in the gas phase with high-energy electron beams. The electron beams displace an electron from the molecule and leave behind a...
8.9K
Tandem Mass Spectrometry01:21

Tandem Mass Spectrometry

2.5K
Tandem mass spectrometry is a technique that uses multiple mass analyzers in series to obtain a higher selectivity and reduce chemical noise during analyte detection. Instruments with multiple analyzers separated by an interaction cell enable secondary fragmentation and selected study of the fragment ions.Secondary fragmentations occur in the interaction cell and can be induced by various factors. Fragmentation induced by collision with inert gases, such as N2, Ar, He, etc., is called...
2.5K
Mass Spectrometry: Isotope Effect01:13

Mass Spectrometry: Isotope Effect

4.3K
Most elements exist in nature as a mixture of isotopes. The isotopes differ in weight due to their respective number of neutrons. The molecular weight of a molecule is different depending on the specific isotope of its elements involved. As a result, the mass spectrum of the molecule exhibits peaks from the same fragment at multiple positions. The positions of these mass signals depend on the mass differences between isotopes. Furthermore, the intensity of these signals is dependent on the...
4.3K
Mass Spectrometry of Amines01:15

Mass Spectrometry of Amines

5.4K
In mass spectroscopy, amines undergo fragmentation to give parent ions with odd molecule weights. This observed mass spectrum follows the nitrogen rule; a molecule with an odd number of nitrogen atoms produces a molecular ion with an odd molecular weight. Amines undergo fragmentation through α cleavage, producing nitrogen-containing cations—iminium ions—and alkyl radicals. Mass spectra of aromatic and cyclic aliphatic amines exhibit strong molecular ion peaks, but acyclic...
5.4K

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Related Experiment Video

Updated: Feb 4, 2026

Sample Preparation Strategies for Mass Spectrometry Imaging of 3D Cell Culture Models
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Sample Preparation Strategies for Mass Spectrometry Imaging of 3D Cell Culture Models

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High-Throughput Enzyme Kinetics with 3D Microfluidics and Imaging SAMDI Mass Spectrometry.

Jennifer Grant, Sohrab Habibi Goudarzi, Milan Mrksich

    Analytical Chemistry
    |September 27, 2018
    PubMed
    Summary
    This summary is machine-generated.

    This study presents a novel microfluidic device for high-throughput enzyme reactions, enabling precise analysis of 2592 distinct reactions with minimal reagent. It introduces imaging self-assembled monolayers for mass spectrometry (iSAMDI-MS) to quantify reaction kinetics.

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    Area of Science:

    • Biochemistry
    • Analytical Chemistry
    • Microfluidics

    Background:

    • Microfluidic systems offer precise reagent control and reduced consumption in biological assays.
    • Optical detection methods are limited to fluorescent or UV-active compounds, and 2D designs restrict experimental flexibility.
    • Existing high-throughput screening methods often require substantial reagent volumes.

    Purpose of the Study:

    • To develop a microfluidic device capable of performing a large number of distinct enzyme reactions with minimal reagent.
    • To introduce a novel detection method, imaging self-assembled monolayers for matrix-assisted laser desorption/ionization mass spectrometry (iSAMDI-MS), for reaction monitoring.
    • To enable quantitative characterization of enzyme kinetics, including Michaelis constant (Km) determination, within the microfluidic system.

    Main Methods:

    • Design and fabrication of a microfluidic device with three inputs for parallel reaction execution.
    • Utilization of imaging self-assembled monolayers for matrix-assisted laser desorption/ionization mass spectrometry (iSAMDI-MS) to immobilize and image reaction products.
    • Quantitative analysis of immobilized products to calculate kinetic parameters like Michaelis constant (Km).

    Main Results:

    • Successfully performed 2592 distinct enzyme reactions using only 150 μL of reagent.
    • Demonstrated quantitative characterization of reaction progress and determination of kinetic parameters.
    • Developed a method to map reaction products onto the microfluidic channel floor for imaging mass spectrometry analysis.

    Conclusions:

    • The developed microfluidic system significantly enhances throughput for enzyme reaction screening.
    • iSAMDI-MS provides a powerful tool for visualizing and quantifying reaction products in microfluidic devices.
    • This integrated approach expands the capabilities of microfluidic detection and high-throughput reaction systems.