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cometchip 96-well system (Trevigen)

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Trevigen cometchip 96-well system
Cometchip 96 Well System, supplied by Trevigen, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/cometchip 96-well system/product/Trevigen
Average 90 stars, based on 1 article reviews

cometchip 96-well system - by Bioz Stars, 2026-05

90/100 stars

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Development of the 384-well CometChip. ( A ) Top left: a 96-well CometChip former (Bio-Techne); Top right: an outline image of the entire 96-well CometChip; Bottom left: an image of comets in four wells of a 96-well CometChip; Bottom right: images of comets within one well of a 96-well CometChip. ( B ) Top left: a 384-well CometChip former; Top right: an outline image of the entire 384-well CometChip; Bottom left: an image of comets in 16 wells of a 384-well CometChip; Bottom right: images of comets within one well of a 384-well CometChip.

Journal: NAR Genomics and Bioinformatics

Article Title: A high-throughput 384-well CometChip platform reveals a role for 3-methyladenine in the cellular response to etoposide-induced DNA damage

doi: 10.1093/nargab/lqac065

Figure Lengend Snippet: Development of the 384-well CometChip. ( A ) Top left: a 96-well CometChip former (Bio-Techne); Top right: an outline image of the entire 96-well CometChip; Bottom left: an image of comets in four wells of a 96-well CometChip; Bottom right: images of comets within one well of a 96-well CometChip. ( B ) Top left: a 384-well CometChip former; Top right: an outline image of the entire 384-well CometChip; Bottom left: an image of comets in 16 wells of a 384-well CometChip; Bottom right: images of comets within one well of a 384-well CometChip.

Article Snippet: The 96-well CometChip system (Bio-Techne) is comprised of hardware for macrowell formation (96-well former) wherein a specially designed bottomless 96-well plate is magnetically compressed onto the agarose.

Techniques:

Comparison of the sensitivity of the 384-well CometChip platform to that of the 96-well CometChip platform for DNA damage evaluation. ( A ) TK6 cells were treated with etoposide (0–10 μM), with a 2 μM ascendance, for 30 min and the level of DNA damage was analyzed using either the 384-well platform or the 96-well platform. The number of comets within each well in a 96-well CometChip or in a 384-well CometChip is shown. Each dot represents the number of the counted comets in a well of the 96-well or 384-well CometChip. ( B ) The DNA damage (%Tail DNA) measured in cells after etoposide exposure was quantified by the 96-well CometChip assay or the 384-well CometChip assay. Each dot represents the average DNA damage level (%Tail DNA) of a well of the 96-well CometChip or the 384-well CometChip. ( C ) TK6 cells were treated with etoposide (10 μM) for 30 min or treated with etoposide (10μM) for 30 min followed by a washout of etoposide by 10ml 1x PBS then re-seeded the cells back into growth medium to repair DNA damage induced by etoposide for 15, 30, 45 or 75 min. Subsequently, the DNA damage (%Tail DNA) was quantified by the 384-well CometChip assay. Each dot represents the average DNA damage level (%Tail DNA) of a well of the 384-well CometChip. DMSO treatment was used as the negative control.

Journal: NAR Genomics and Bioinformatics

Article Title: A high-throughput 384-well CometChip platform reveals a role for 3-methyladenine in the cellular response to etoposide-induced DNA damage

doi: 10.1093/nargab/lqac065

Figure Lengend Snippet: Comparison of the sensitivity of the 384-well CometChip platform to that of the 96-well CometChip platform for DNA damage evaluation. ( A ) TK6 cells were treated with etoposide (0–10 μM), with a 2 μM ascendance, for 30 min and the level of DNA damage was analyzed using either the 384-well platform or the 96-well platform. The number of comets within each well in a 96-well CometChip or in a 384-well CometChip is shown. Each dot represents the number of the counted comets in a well of the 96-well or 384-well CometChip. ( B ) The DNA damage (%Tail DNA) measured in cells after etoposide exposure was quantified by the 96-well CometChip assay or the 384-well CometChip assay. Each dot represents the average DNA damage level (%Tail DNA) of a well of the 96-well CometChip or the 384-well CometChip. ( C ) TK6 cells were treated with etoposide (10 μM) for 30 min or treated with etoposide (10μM) for 30 min followed by a washout of etoposide by 10ml 1x PBS then re-seeded the cells back into growth medium to repair DNA damage induced by etoposide for 15, 30, 45 or 75 min. Subsequently, the DNA damage (%Tail DNA) was quantified by the 384-well CometChip assay. Each dot represents the average DNA damage level (%Tail DNA) of a well of the 384-well CometChip. DMSO treatment was used as the negative control.

Techniques: Comparison, Negative Control

96-well CometChip assay validation of the 3-MA increase in the etoposide-induced DNA damage. ( A ) TK6 cells were pre-treated with the top 5 kinase inhibitor hits for 30 min followed by etoposide (2 or 4 μM) for an additional 30 min. The DNA damage was analyzed using a 96-well platform (** P < 0.01, *** P < 0.001). ( B ) TK6 cells were pre-treated with 3-MA at 10 or 20 μM for 1 or 2 h followed by etoposide (2 μM) for an additional 30 min. The DNA damage was analyzed using a 96-well platform (**** P < 0.0001).

Journal: NAR Genomics and Bioinformatics

Article Title: A high-throughput 384-well CometChip platform reveals a role for 3-methyladenine in the cellular response to etoposide-induced DNA damage

doi: 10.1093/nargab/lqac065

Figure Lengend Snippet: 96-well CometChip assay validation of the 3-MA increase in the etoposide-induced DNA damage. ( A ) TK6 cells were pre-treated with the top 5 kinase inhibitor hits for 30 min followed by etoposide (2 or 4 μM) for an additional 30 min. The DNA damage was analyzed using a 96-well platform (** P < 0.01, *** P < 0.001). ( B ) TK6 cells were pre-treated with 3-MA at 10 or 20 μM for 1 or 2 h followed by etoposide (2 μM) for an additional 30 min. The DNA damage was analyzed using a 96-well platform (**** P < 0.0001).

Techniques:

Analysis of known DNA damaging agents. ( A ) Etoposide is shown to induce replication-dependent DSBs and cause significant DNA damage at concentrations greater than 1 μM (n = 1069–1374). Shown is the plot of % tail DNA for TK6 and Jurkat cells following exposure (60 min) to the agent at the doses indicated. ( B ) H 2 O 2 did not cause DNA damage below 1 μM, with elevated levels of DNA damage observed at 50 μM (n = 450–1200). Shown is the plot of % tail DNA for TK6 and Jurkat cells following exposure (60 min) to the agent at the doses indicated. ( C ) MNNG, a potent SN1 DNA alkylating agent , was the most genotoxic of the compounds tested and caused the maximum amount of DNA damage that could be measured at 10 μM (n = 1075–1374). Shown is the plot of % tail DNA for TK6 and Jurkat cells following exposure (60 min) to the agent at the doses indicated. ( D ) MMS, an SN2 DNA alkylating agent , induced only modest DNA damage on the cells tested, with significant DNA damage observed only at concentrations above 50 μM (n = 812–1294). All assays were conducted in duplicate. Error bars represent mean ± 95% CI. Shown is the plot of % tail DNA for TK6 and Jurkat cells following exposure (60 min) to the agent at the doses indicated. For each panel, the images below the plot are representative CometChip images at the doses indicated.

Journal: Scientific Reports

Article Title: Next generation high throughput DNA damage detection platform for genotoxic compound screening

doi: 10.1038/s41598-018-20995-w

Figure Lengend Snippet: Analysis of known DNA damaging agents. ( A ) Etoposide is shown to induce replication-dependent DSBs and cause significant DNA damage at concentrations greater than 1 μM (n = 1069–1374). Shown is the plot of % tail DNA for TK6 and Jurkat cells following exposure (60 min) to the agent at the doses indicated. ( B ) H 2 O 2 did not cause DNA damage below 1 μM, with elevated levels of DNA damage observed at 50 μM (n = 450–1200). Shown is the plot of % tail DNA for TK6 and Jurkat cells following exposure (60 min) to the agent at the doses indicated. ( C ) MNNG, a potent SN1 DNA alkylating agent , was the most genotoxic of the compounds tested and caused the maximum amount of DNA damage that could be measured at 10 μM (n = 1075–1374). Shown is the plot of % tail DNA for TK6 and Jurkat cells following exposure (60 min) to the agent at the doses indicated. ( D ) MMS, an SN2 DNA alkylating agent , induced only modest DNA damage on the cells tested, with significant DNA damage observed only at concentrations above 50 μM (n = 812–1294). All assays were conducted in duplicate. Error bars represent mean ± 95% CI. Shown is the plot of % tail DNA for TK6 and Jurkat cells following exposure (60 min) to the agent at the doses indicated. For each panel, the images below the plot are representative CometChip images at the doses indicated.

Article Snippet: The CometChip Platform is now available from Trevigen, a division of Bio-Techne (Minneapolis, MN) and includes the disposable 30 μM CometChips, the CometChip 96-well magnetically sealed cassettes (formers), the CometChip Electrophoresis System (CES) and the Comet Analysis Software (CAS).

Techniques:

Design of Next-Generation CometChip hardware and work-flow. Close up of the CometChip showing the agarose surface chemically bound on a glass support. The CometChip is inserted into a Former base. When assembled, the Former allows each of the 96 wells in the CometChip to be treated individually. Each well has, in turn, more than 500 microwells that enable cells to be gravity loaded into the CometChip, the result is that all cells are on a single focal plane. The dedicated electrophoresis system limits the field variability by reducing the distance between the two electrodes. After electrophoresis, the CometChip is stained and images are acquired using an automated image cytometer (Celigo). These images are subsequently analyzed using dedicated CAS optimized for the CometChip high throughput methodology.

Journal: Scientific Reports

Article Title: Next generation high throughput DNA damage detection platform for genotoxic compound screening

doi: 10.1038/s41598-018-20995-w

Figure Lengend Snippet: Design of Next-Generation CometChip hardware and work-flow. Close up of the CometChip showing the agarose surface chemically bound on a glass support. The CometChip is inserted into a Former base. When assembled, the Former allows each of the 96 wells in the CometChip to be treated individually. Each well has, in turn, more than 500 microwells that enable cells to be gravity loaded into the CometChip, the result is that all cells are on a single focal plane. The dedicated electrophoresis system limits the field variability by reducing the distance between the two electrodes. After electrophoresis, the CometChip is stained and images are acquired using an automated image cytometer (Celigo). These images are subsequently analyzed using dedicated CAS optimized for the CometChip high throughput methodology.

Techniques: Electrophoresis, Staining, Cytometry, High Throughput Screening Assay

Acquisition and analysis of image data. ( A ) Image acquisition and comet analysis using dedicated CAS. Representative image of a single CometChip well after DNA damage treatment. Image is comprised of 16 individual post-acquisition stitched images. The insert shows the region expanded in ( B ). ( B ) Higher magnification of the insert from ( A ), shows individual wells. The CAS automated analysis software detects and boxes each comet and colors the comet based on the intensity of the signal. ( C ) Two examples of comets post-CAS analysis – ( upper ) negative control comet with endogenous level of DNA damage. The DNA does not migrate significantly due to supercoiling. ( lower ) Example of a comet from a highly-damaged cell that has been exposed to etoposide. In each example, the white line at the bottom of the image marks the beginning of the comet head, the red line marks the outermost edge of the comet head and the green line marks the end of the comet tail.

Journal: Scientific Reports

Article Title: Next generation high throughput DNA damage detection platform for genotoxic compound screening

doi: 10.1038/s41598-018-20995-w

Figure Lengend Snippet: Acquisition and analysis of image data. ( A ) Image acquisition and comet analysis using dedicated CAS. Representative image of a single CometChip well after DNA damage treatment. Image is comprised of 16 individual post-acquisition stitched images. The insert shows the region expanded in ( B ). ( B ) Higher magnification of the insert from ( A ), shows individual wells. The CAS automated analysis software detects and boxes each comet and colors the comet based on the intensity of the signal. ( C ) Two examples of comets post-CAS analysis – ( upper ) negative control comet with endogenous level of DNA damage. The DNA does not migrate significantly due to supercoiling. ( lower ) Example of a comet from a highly-damaged cell that has been exposed to etoposide. In each example, the white line at the bottom of the image marks the beginning of the comet head, the red line marks the outermost edge of the comet head and the green line marks the end of the comet tail.

Techniques: Software, Negative Control

CometChip validation. ( A ) Intra-assay variability. The wells on the outer edge of the plate had the most variability compared to controls. Overall, we observed average variability of less than 4.2% of mean difference and maximum variability less than 10% of mean difference. The average group value is depicted as a red dotted line. Error bars = mean with 95% CI (n = 644–749). ( B ) Inter-assay variability. Cells were treated with etoposide (5 μM) and DNA damage was measured after 1 h. Each point represents a separate experiment (n = 17 per cell line). CV was measured at 13% for the JK cells and 11% for the TK6 cells (n = 420–580 per point). Error bars = mean ± SD. Statistical analysis was conducted via a two-sided Student’s t-test. ( C ) Linear DNA damage response to the DSB causing agent, etoposide. Assay was able to resolve differences in DNA damage caused by 1 μM increments of the compound (****p < 0.0001, n = 1000–2000 comets; r = 0.98). Statistical analysis was conducted via a two-sided Student’s t-test.

Journal: Scientific Reports

Article Title: Next generation high throughput DNA damage detection platform for genotoxic compound screening

doi: 10.1038/s41598-018-20995-w

Figure Lengend Snippet: CometChip validation. ( A ) Intra-assay variability. The wells on the outer edge of the plate had the most variability compared to controls. Overall, we observed average variability of less than 4.2% of mean difference and maximum variability less than 10% of mean difference. The average group value is depicted as a red dotted line. Error bars = mean with 95% CI (n = 644–749). ( B ) Inter-assay variability. Cells were treated with etoposide (5 μM) and DNA damage was measured after 1 h. Each point represents a separate experiment (n = 17 per cell line). CV was measured at 13% for the JK cells and 11% for the TK6 cells (n = 420–580 per point). Error bars = mean ± SD. Statistical analysis was conducted via a two-sided Student’s t-test. ( C ) Linear DNA damage response to the DSB causing agent, etoposide. Assay was able to resolve differences in DNA damage caused by 1 μM increments of the compound (****p < 0.0001, n = 1000–2000 comets; r = 0.98). Statistical analysis was conducted via a two-sided Student’s t-test.

Techniques: Biomarker Discovery, Intra Assay, Inter Assay

National Toxicology Program 74 compound plate – agents scored as damaging by CometChip (in descending order by mean % tail DNA within each category). (P value was calculated using a one tailed Student’s t-test).

Journal: Scientific Reports

Article Title: Next generation high throughput DNA damage detection platform for genotoxic compound screening

doi: 10.1038/s41598-018-20995-w

Figure Lengend Snippet: National Toxicology Program 74 compound plate – agents scored as damaging by CometChip (in descending order by mean % tail DNA within each category). (P value was calculated using a one tailed Student’s t-test).

Techniques: Significance Assay, One-tailed Test

National Toxicology Program 74 compound plate – agents scored as non-damaging by CometChip (in descending order by mean % tail DNA within each category).

Journal: Scientific Reports

Article Title: Next generation high throughput DNA damage detection platform for genotoxic compound screening

doi: 10.1038/s41598-018-20995-w

Figure Lengend Snippet: National Toxicology Program 74 compound plate – agents scored as non-damaging by CometChip (in descending order by mean % tail DNA within each category).

Techniques:

27 known genotoxic agents in the NTP plate and results of the CometChip pre-screen (100 µM concentration, 30-min exposure duration).

Journal: Scientific Reports

Article Title: Next generation high throughput DNA damage detection platform for genotoxic compound screening

doi: 10.1038/s41598-018-20995-w

Figure Lengend Snippet: 27 known genotoxic agents in the NTP plate and results of the CometChip pre-screen (100 µM concentration, 30-min exposure duration).

Techniques: Concentration Assay, In Vitro, Single Cell Gel Electrophoresis

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