Scratch Wound Assay

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  Cell health and viability measurements provide essential insight into a broad range of biological processes and treatment responses. Diverse assay formats and reagents have been developed that measure specific aspects of cell viability corresponding to particular cellular response pathways and mechanisms of injury.
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  Cellular metabolism comprises a number of biochemical reactions that occur in concert within the cells of living organisms. These orchestrated reactions, better described as metabolic pathways allow organisms to function and respond to environmental changes. Agilent cell metabolism assays detect discrete changes in cell bioenergetics in real time, providing a window into the critical functions that provide ATP, the energy that cells need for activation, signaling, proliferation, and biosynthesis.
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  Cell migration and invasion are live cell kinetic assays useful in oncology research. The principle difference between migration and invasion is that migration refers to normal cell movement; whereas invasion describes cells actively invading surrounding tissue. It can do this under the influence of a chemoattractant gradient and secretion of proteases. There are numerous methods for performing these assays including scratch assays performed on a monolayer of cells adhered to plasticware (microplates or culture inserts) or using 3D cell culture models.
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- Endotoxin Analysis
  Bacterial endotoxins are lipopolysaccharides (LPS) generally found in the outer membrane of gram-negative bacteria, such as E.coli, and are widely present in our environment. Because of the pyrogenic nature of endotoxins, with human effects from fever to hemorrhagic stroke, FDA regulations require that raw materials (e.g. water) used in parenteral drugs be tested for endotoxin levels.
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- Immunology
  The immune system relies on diverse mechanisms working in concert to defend the host from infection and to identify and remove aberrant or damaged cells. Dysregulation of the immune system can lead to a variety of disease conditions, including autoimmune disorders, allergies, and cancer. Additionally, the immune system is a critical consideration for vaccine development and cell, tissue and organ transplants.
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  While harnessing the patient’s immune system to fight cancer is not a new concept, there have been significant strides in the past decade to more effectively direct and activate components of the immune system and counter the measures certain tumors adopt to shield itself from immune attack. These include immunotherapeutic drugs based on directing immune cells to tumors based on specific antigens they present, PD-1 checkpoint inhibition and the ex vivo manipulation of the patient’s own immune cells to better drive an immune response to cancer.
- Microbiology
  Since the advent of a lens system that enabled the visualization of bacteria and protists in 1671, the microscope remains a ubiquitous tool for the scientific study of live and fixed microorganisms. Investigating the structure, function, and typing of the highly diverse array of microorganisms spans heterogeneous applications including, but not limited to, the elucidation, diagnosis, and treatment of human and plant disease, food and water safety testing, vaccine development, investigation and production of industrial lipases, protein transfection to mammalian cells, environmental health and impact appraisal, horticultural and agricultural soil assessment, and exploration of alternative fuel sources.
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- Rapid Cell Signaling
  G-protein coupled, seven-transmembrane (7-TM) receptors (GPCRs) represent large, diverse gene families found in fungi, plants and animals. The GPCR family of receptors is involved primarily with internalizing extracellular environmental signals via interaction with intracellular G-proteins. GPCR ligands and modulators range from peptide and non-peptide molecules to chemokines, nucleosides, growth factors and light.
- Stem Cell Research
  Research into the reprogramming of somatic cells into pluripotent stem cells received the Nobel Prize in 2012. The long term importance of this research resides in the use of stem cells for regenerative therapies; for the short term, however these cells represent a potential unlimited source of highly physiologically relevant cells, akin to human primary cells, that can be used as drug discovery reagents.
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The wound healing or scratch assay is one of the most highly used in vitro methods to monitor and quantify collective cell migration. The Agilent BioTek AutoScratch wound making tool creates uniform and precise wounds in either 24- or 96-well plates. Cell migration into the wound area over time is monitored within the Agilent BioTek Lionheart FX automated live cell imager or Agilent BioTek Cytation cell imaging multimode reader. The Agilent BioTek Gen5 microplate reader and imager software analysis delivers detailed and reproducible cell migration results.

AutoScratch: Automated Wound Creation for Scratch Wound Healing Assays

Automated wound creation decreases variability within and between wounds, improving repeatability of cell migration data. Combined with kinetic image capture and analysis, the Agilent BioTek Scratch Assay Starter Kit delivers a complete automated solution for conducting robust kinetic scratch wound assays.

Autoscratch

Figure 1. The AutoScratch wound making tool can automatically create repeatable scratch wounds.

cell-migration-invasion

Figure 2. Clean: Walk-away procedure cleans and sterilizes pins before and after wound creation.

Wound: Repeatable wounds created automatically in 24- or 96-well plates with the push of a button.

Scratch Wound Assay Procedure

Cell Migration Assay: 3D Bioprinting Visual Abstract
Step 1. Capture: Kinetic high contrast brightfield (HCBF) images are captured based on desired interval and duration.

Step 2. Analyze: Images are processed and analyzed to measure cell migration in real-time.

Label-Free Tumor Invasion
Step 3. Report: Kinetic profiles of wound closure rates reveal detailed information on cell migration across treatment conditions.

Increase assay throughput and performance

Agilent BioTek BioSpa compatible assay performs live cell analysis on up to 8 microplates. The BioSpa live cell imaging system enables multiplate kinetic analysis of the scratch wound assay.
BioSpa

Liquid handling automation is achieved with the Agilent BioTek MultiFlo FX multimode dispenser.

Resources

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For Research Use Only. Not for use in diagnostic procedures.

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