Hi, what would bethe outcome measured in this assay? The slope of a plot of area coverd by cell vs time, or The decrease of the area of the wound at a certain timepoint? Thank you very much!
I got this information from you elsewhere: "What I normally do is that I draw a line across all wells with a black marker prior to seeding cells (this can also be done after cells have attached and media is removed). On the day of the assay, I scratch two or three lines with a 200ul tip perpendicular to the black marker line. By doing this, the black line is my point of reference and I image the wound right below or above the marker line and label accordingly (Right wound below line would be RB for right bottom). Best, Olga Soto"
2 ปีที่แล้ว +1
Here are my two cents to this matter. If you follow the procedure indicated by Olga (see below), it will work out just fine, probably even better than using the standard method. The important thing is to have a proper reference to where the images were captured at the beginning of the experiment, so that you can compare the initial gap with the one obtained at the end of the experiment. A major source of error in this type of experiments is making measurements at different positions. Some of the imaging software available nowadays allow you to calculate the "empty" area rather than just the distance between the edges where the cells are located. This is probably better simply because it allows for more accurate measurements, including cells that migrate apart from the main body of cells. The area occupied by those cells is taken off from the "empty" area. Combining this approach with the use of the reference lines described by Olga are likely to give the most accurate measurements. Thank you for your question!
To accelerate wound healing a no contact dressing works well. It is called Podophylus antimicrobial Insole and DermaDomus, both allow oxygen in the wound…
I have a question: she says around 8 minutes, "Our vehicle control will be DMSO" but the subtitle instructions say, "add negative control DMSO." Is DMSO both the vehicle and the negative control? Thank you!
4 ปีที่แล้ว +7
Maya, DMSO is an additional control that is needed simply because the compound being tested was dissolved using DMSO (many chemicals are not soluble in water and require the use of an organic solvent such as DMSO). So, since the compound was dissolved in DMSO, you need to test to see whether DMSO by itself has any effect. The negative control should be NO TREATMENT, meaning nothing is added at all, so the cells are kept in normal culture medium without the addition of any additional compound or solvent. This allows you to determine whether the cells are capable of any migration whatsoever (not all cells migrate). It also allows you to determine how much they would migrate under normal conditions. I hope this helps.
![FIJI (ImageJ): Analysis of Scratch Assays, Wound Healing & Cell Migration [Wound Healing Size Tool]](/img/n.gif)
I have a question, did you do two scrapes with tip? why?
I have the same question..
Many thanks for your information. I'm curious about how can we record the video cell proliferation process in the incubator?
Hi, what would bethe outcome measured in this assay?
The slope of a plot of area coverd by cell vs time, or
The decrease of the area of the wound at a certain timepoint?
Thank you very much!
Many thanks for this video. However, will there be any difference if you had drawn the reference lines before ever plating the cells?
I got this information from you elsewhere: "What I normally do is that I draw a line across all wells with a black marker prior to seeding cells (this can also be done after cells have attached and media is removed). On the day of the assay, I scratch two or three lines with a 200ul tip perpendicular to the black marker line. By doing this, the black line is my point of reference and I image the wound right below or above the marker line and label accordingly (Right wound below line would be RB for right bottom).
Best,
Olga Soto"
Here are my two cents to this matter. If you follow the procedure indicated by Olga (see below), it will work out just fine, probably even better than using the standard method. The important thing is to have a proper reference to where the images were captured at the beginning of the experiment, so that you can compare the initial gap with the one obtained at the end of the experiment. A major source of error in this type of experiments is making measurements at different positions. Some of the imaging software available nowadays allow you to calculate the "empty" area rather than just the distance between the edges where the cells are located. This is probably better simply because it allows for more accurate measurements, including cells that migrate apart from the main body of cells. The area occupied by those cells is taken off from the "empty" area. Combining this approach with the use of the reference lines described by Olga are likely to give the most accurate measurements. Thank you for your question!
which cell line u used in this video?
To accelerate wound healing a no contact dressing works well. It is called Podophylus antimicrobial Insole and DermaDomus, both allow oxygen in the wound…
I have a question: she says around 8 minutes, "Our vehicle control will be DMSO" but the subtitle instructions say, "add negative control DMSO." Is DMSO both the vehicle and the negative control? Thank you!
Maya, DMSO is an additional control that is needed simply because the compound being tested was dissolved using DMSO (many chemicals are not soluble in water and require the use of an organic solvent such as DMSO). So, since the compound was dissolved in DMSO, you need to test to see whether DMSO by itself has any effect. The negative control should be NO TREATMENT, meaning nothing is added at all, so the cells are kept in normal culture medium without the addition of any additional compound or solvent. This allows you to determine whether the cells are capable of any migration whatsoever (not all cells migrate). It also allows you to determine how much they would migrate under normal conditions. I hope this helps.
thank u
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