CometChip®

High Throughput DNA Damage Analysis

µM Etoposide % DNA in Tail SD CV

0.00 5.03 0.89 17.61

1.25 21.47 1.21 5.64

2.50 37.33 2.63 7.04

5.00 57.70 4.18 7.24

µM Etoposide % DNA in Tail SD CV

0.00 5.43 1.05 19.33

1.25 19.78 1.91 9.68

2.50 32.89 1.97 5.99

5.00 59.65 3.77 6.32

Introduction to CometChip®

The comet assay has traditionally been used due to its high sensitivity and simplicity, but the original format is a laborious and a time-consuming hunt to find non-overlapping cells for analysis.

The CometChip® Platform provides high density data with minimal overlaps by capturing cells into an array of micropores. Scan and analyze over 14,000 comets in 10-15 minutes with R&D Systems’ CometAssay® Analysis Software which automatically locates and scores comets in 4X or 10X well images for instant and unbiased analysis.

Unique Features• Developed and Manufactured under ISO 9001/2008 guidelines

• Only Integrated and Standardized System

• Up to 400 non-overlapping comets per well for analysis

• Lower variance between runs

• Comparable results to traditional comet assay

• Versatility for multiple cell types

• Reproducible Data

• Cost Savings with 10,000% increase in throughput

Lower Variance Between Runs

<10% Variability Between CometChips with treated Samples

Low Chip to Chip CVs • Meets USP guidelines

• Meets ASCLS specification

Unbiased Analysis• No overlapping Comets

• Minimal editing

Inter (Different Day) Run Variabilityn=3 CometChips

Intra (Same Day) Run Variabilityn=3 CometChips

Instant and Unbiased Analysis

- Up to 400 Comets in a single well

- Up to 200 Comets per 4X image

2Weeks of Work Compressed into Days

Comparable Results to Traditional Comet Assay

Comparable Results & Versatility1 CometChip versus 30 slides

Versatility for Multiple Cell Types

Reproducible Data

% T

ail D

NA,

n=3

CometChip®

SlidesCometChip®

Slides

IR Dose (Gy)

Alkaline - SSB Neutral - DSB

IR Dose (Gy)

Tail

Leng

th (μ

m),

n=3

0 2 4 6 8 10 0 20 40 60 80 100

60

40

20

0

120

100

80

60

40

Human Lymphoblast

% T

ail D

NA

IR Dose (Gy)

% Tail D

NA

Ovarian Carcinoma Primary Hepatocyte

0 2 4 6 8 10 0 2 4 6 8 10 0 2 4 6 8 10

80

60

40

20

0

80

60

40

20

0

% D

NA

in T

ail (

n=3)

HepaRG: 4 hr MMS treatment

μg/ml MMS

Operator AOperator B

100

90

80

70

60

50

40

30

20

10

00 10 20 30 40 50 60 70 80

Separate experiments performed by different operators on the same day

• HepaRG cells treated with increasing concentration of MMS.

• Two separate experiments performed by different operators on the same day.

• Each concentration performed in triplicate.

Wood DK, Weingeist DM, Bhatia SN, and Engelward BP. 2010.Single cell trapping and DNA damage analysis using microwell arrays. PNAS 107(22): 10008-10013

A Complete CometChip® Platform

CometChip®, 30 micron (catalog # 4260-096-01)Agarose with patterned micropores adhere to a glass slide. Designed to allow single cells to be analyzed with no overlap.

CometAssay® Analysis Software (catalog # 4260-00-CS)Automatically locate and score comets from 4X or 10X well images to allow single cells to be analyzed with no overlap.

96-Well CometChip® System (catalog # 4260-096-CS)Macrowell former creates 96 individual wells on the CometChip for ease of use.

CometAssay® Electrophoresis System II (catalog # 4250-050-ES)Optimize and standardize electrophoresis of the CometChip.

CometChip® Kit (catalog # 4260-096-K)Qualified reagents and optimized protocols for reproducible results.

Agarose withPatterned

Micropores

ActivatedGlass Slide

CometChip®

Scan Analyze

Edit

Single Well Image

Learn more | rndsystems.com/products/cometassay

Cell Loading and Treatment

~400 Micropores per Well

CometChip® Platform Kits

Frequently Asked Questions

CometChip® Custom Services for Toxicology Studies

Is it possible to use a multi-channel pipette when loading or treating samples in the 96-Well CometChip® System? Yes, the system is designed for multi-channel pipettor.

What are the throughput capabilities with the complete platform?Using the CometAssay Electrophoresis System II, the slide tray can hold 3 CometChips per run resulting in up to 115,200 comets that can simultaneously be analyzed in the CometAssay Analysis Software Program, within a 2-day experiment run.

I noticed the CometChip® is not designed for overnight treatment. Is it possible to seed the cells/treat in normal 96 well tissue culture plate and transfer? Yes, many customers perform cell treatment in tissue culture plates and then transfer to 96-Well CometChip System. It only takes 20-30 minutes for cells to load into the micropores of each well.

Is the CometChip® available with other pore sizes?Yes, inquire about 20 micron and 40 micron options, available for custom ordering.

How do I improve the efficiency of cell loading into the micropores of the CometChip®? To improve loading, filter cells using 40-75 micron filter. Cells can generally be loaded without counting if between 1-10 x 10^5 cells/ml. There is no harm in increasing the loading time to 30-60 minutes or cell concentration. It is helpful to rock the macroformer after the first 10-15 minutes to promote loading efficiency.

Is the CometChip® available with other matrices?Yes, inquire about collagen CometChips. Available for custom ordering.

Can the CometChip® be used with different electrophoresis systems? Conditions have been optimized using R&D Systems’ CometAssay Electrophoresis System II to generate reproducible results.

Are changes documented in the CometAssay® Analysis Software? Yes, all edits performed during comet analysis are documented.

Is the CometAssay® Analysis Software Mac compatible? No, the software is designed for PC to work under Windows 10, 8/8.1 and 7.

Can the CometAssay® Analysis Software import all well images simultaneously? No, this would limit customer flexibility in the number of images per well. The Well Explorer platform offers the simple “Drag and Drop” of files into slide wells within minutes.

How many people can use the software? The software is purchased as a single user license, however multiple license pricing is available.

Do you have additional training material for the CometChip platform?Additional information and training can be found on our CometAssay web page, ndsystems.com/products/cometassay.

CometChip® Kit (catalog # 4260-096-K)

CometChip® Starter Kit (catalog # 4260-096-CSK)

CometChip® Electrophoresis Starter Kit (catalog # 4260-096-ESK)

Component Amount Catalog #

CometChip®, 30 micron 1 each 4260-096-01

Lysis Solution 100 ml 4250-010-01

CometAssay® LMAgarose 15 ml 4250-050-02

Component Amount Catalog #

CometChip® Kit 1 each 4260-096-K

96-Well CometChip® System 1 system 4260-096-CS

Component Amount Catalog #

CometChip® Starter Kit 1 each 4260-096-CSK

CometAssay® Electrophoresis System II 1 system 4250-050-ES

Utilize our in-house CometChip expertise to propel your toxicology project forward. Consult with our expert scientists to design your next project.

CometChip Custom Services Include:

• Custom micropore size

• Collagen matrix CometChips

• Customer sample testing services

• Data analysis using our CometAssay Analysis Software

Learn More: rndsystems.com/services

BR_Comet Assay_RSDFY20-13178

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For research use or manufacturing purposes only. Trademarks and registered trademarks are the property of their respective owners.

References Using CometChip®

Wood, David K. et al. “Single Cell Trapping and DNA Damage Analysis Using Microwell Arrays.” Proceedings of the National Academy of Sciences of the United States of America 107.22 (2010): 10008–10013.

Seo, JE et al., “Quantitative comparison of in vitro genotoxicity between metabolically competent HepaRG cells and HepG2 cells using the high-throughput high-content CometChip assay” Arch Toxicol. (2019)

Ngo, LN et al., “Ultrasensitive CometChip Assay for Screening Potentially Carcinogenic DNA Adducts by Trapping DNA Repair Intermediates” Nucleic Acids Research (2019)

Sykora, Peter et al. “Next Generation High Throughput DNA Damage Detection Platform for Genotoxic Compound Screening.” Scientific Reports 8:2771 (2018): 1-20.

Ge, Jing et al. “Micropatterned Comet Assay Enables High Throughput and Sensitive DNA Damage Quantification.” Mutagenesis 30.1 (2015): 11–19.

Li, Na et al. “Influenza Infection Induces Host DNA Damage and Dynamic DNA Damage Responses during Tissue Regeneration.” Cellular and molecular life sciences : CMLS 72.15 (2015): 2973–2988.

Ge, Jing et al. “CometChip: A High-Throughput 96-Well Platform for Measuring DNA Damage in Microarrayed Human Cells.” Journal of Visualized Experiments : JoVE 92 (2014): 50607.

Sotiriou, Georgios A. et al. “Engineering Safer-by-Design, Transparent, Silica-Coated ZnO Nanorods with Reduced DNA Damage Potential.” Environmental science. Nano 1.2 (2014): 144–153.

Watson, Christa et al. “High-Throughput Screening Platform for Engineered NanoparticleMediated Genotoxicity Using CometChip Technology.” ACS Nano 8.3 (2014): 2118–2133.

Ge, Jing et al. “Standard Fluorescent Imaging of Live Cells Is Highly Genotoxic.” Cytometry. Part A : the journal of the International Society for Analytical Cytology 83.6 (2013): 552–560.

Weingeist, David M. et al. “Single-Cell Microarray Enables High-Throughput Evaluation of DNA Double-Strand Breaks and DNA Repair Inhibitors.” Cell Cycle 12.6 (2013): 907–915.

Chao, Ming-Wei et al. “Genotoxicity of 2,6- and 3,5-Dimethylaniline in Cultured Mammalian Cells: The Role of Reactive Oxygen Species.” Toxicological Sciences 130.1 (2012): 48–59.

Mutamba, James T. et al. “XRCC1 and Base Excision Repair Balance in Response to Nitric Oxide.” DNA repair 10.12 (2011): 1282–1293.