Talk:20.109(S07): Measuring calcium in vivo

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Data from the Tuesday/Thursday lab section

Before you leave lab today, please upload at least on image and include a description of it. Write as though you are submitting a figure for a journal article, i.e. include a title for the image (often the conclusion you would like the reader to draw) and some information about the image for readers who know nothing about the experiment you've performed (cell type, magnification, intent of the experiment, conclusion you can draw). Be sure to include your team color as well.

  • T/Th Green - Increased fluorescence in cyproheptadine treated Fluo-3 MES cells (10x mag)

    T/Th Green - Increased fluorescence in cyproheptadine treated Fluo-3 MES cells (10x mag)

  • High fluorescence in CaCl2 treated Fluo-3 MES cells. (Source: TR Purple Team)

    High fluorescence in CaCl2 treated Fluo-3 MES cells. (Source: TR Purple Team)

  • T/R Pink - High fluorescence in EGTA treated Fluo-3 MES cells (400x mag)

    T/R Pink - High fluorescence in EGTA treated Fluo-3 MES cells (400x mag)

  • Positive Control, Mice Embryonic Stem Cells under Fluorescent light with inverse pericam DNA transfected (400x) (Source: TR Red Team)

    Positive Control, Mice Embryonic Stem Cells under Fluorescent light with inverse pericam DNA transfected (400x) (Source: TR Red Team)

  • T/TH Yellow - High fluorescence in Fluo-3 MES cells treated with 1 uM histamine, 400X. (Source: TR Yellow Team)

    T/TH Yellow - High fluorescence in Fluo-3 MES cells treated with 1 uM histamine, 400X. (Source: TR Yellow Team)

  • T/TH BLUE- High Flourescence in Fluo-3 MES cells treated with 1 uM histamine.

    T/TH BLUE- High Flourescence in Fluo-3 MES cells treated with 1 uM histamine.

  • W/F PINK- This picture is of the positive control with the +Fluo-3, AM. The magnification is 100X on mouse embryonic stem cells under FITC lighting. The intent of the procedure was to see the effect of histamine on the flourescence of these cells. Histamine would increase Ca2+ by opening ion channels in the cells and allowing Ca2+ to rush in due to a concentration gradient. This positive control is without the histamine, but flourescence can still be seen due to the free Ca2+ in the cells normally.

    W/F PINK- This picture is of the positive control with the +Fluo-3, AM. The magnification is 100X on mouse embryonic stem cells under FITC lighting. The intent of the procedure was to see the effect of histamine on the flourescence of these cells. Histamine would increase Ca2+ by opening ion channels in the cells and allowing Ca2+ to rush in due to a concentration gradient. This positive control is without the histamine, but flourescence can still be seen due to the free Ca2+ in the cells normally.

  • W/F Purple - "Negative Control of Fluo-3 treated ES Cells exhibit background fluorescence." These are mouse embryonic stem cells magnified at 400x. The intent of the experiment is to note the difference in calcium levels between cells treated with calcium mobilization chemicals (cyproheptadine hydrochloride in our experiment)and those cells not treated. Our fluorescence levels between treated and untreated cells appears to be the same. As expected the cyproheptadine is proved to be selective inhibitor of histamine - and does not act as a calcium chelator.

    W/F Purple - "Negative Control of Fluo-3 treated ES Cells exhibit background fluorescence." These are mouse embryonic stem cells magnified at 400x. The intent of the experiment is to note the difference in calcium levels between cells treated with calcium mobilization chemicals (cyproheptadine hydrochloride in our experiment)and those cells not treated. Our fluorescence levels between treated and untreated cells appears to be the same. As expected the cyproheptadine is proved to be selective inhibitor of histamine - and does not act as a calcium chelator.

  • W/F Yellow - This is a control of Fluo-3 labeled cells, which appear generally healthy, serving as a good comparison against our treated sample.

    W/F Yellow - This is a control of Fluo-3 labeled cells, which appear generally healthy, serving as a good comparison against our treated sample.

  • W/F Yellow - This is a representative sample of cells treated with 1mM CaCl2. In this work, we had hoped to test the specificity of our Calcium sensors to Calcium compared to Magnesium by adding CaCl2 to some cells and MgCl2 to others. This concentration was probably detrimental to the cells, resulting in the withered appearance. Both images are magnified at 400x.

    W/F Yellow - This is a representative sample of cells treated with 1mM CaCl2. In this work, we had hoped to test the specificity of our Calcium sensors to Calcium compared to Magnesium by adding CaCl2 to some cells and MgCl2 to others. This concentration was probably detrimental to the cells, resulting in the withered appearance. Both images are magnified at 400x.

  • W/F Blue - This is a sample of our Fluo-3 treated ES cells with 1 uM Histamine and 50 uM of Ca2+ after a 4-minute incubation magnified at 400X.

    W/F Blue - This is a sample of our Fluo-3 treated ES cells with 1 uM Histamine and 50 uM of Ca2+ after a 4-minute incubation magnified at 400X.

  • W/F Green - These are mouse embryonic stem cells expressing the calcium indicator with 0.2mM Histamine stimulation

    W/F Green - These are mouse embryonic stem cells expressing the calcium indicator with 0.2mM Histamine stimulation


Schedule

Today's lab will require that you arrive in lab at 1/2 hour intervals. Please knock on the door loudly so we can hear you in the cell culture facility.

Tuesday/Thursday Schedule

Team Color arrive in teaching lab at
Green 1:00
Purple 1:30
Red 2:00
Blue 4:00
Pink 2:30
Yellow 3:30


Wednesday/Friday Schedule

Team Color arrive in teaching lab at
Green 1:30
Purple 2:00
Red 2:30
Blue 4:00
Pink 1:00
Yellow 3:30

Data from the Wednesday/Friday lab section

Before you leave lab today, please upload at least on image and include a description of it. Write as though you are submitting a figure for a journal article, i.e. include a title for the image (often the conclusion you would like the reader to draw) and some information about the image for readers who know nothing about the experiment you've performed (cell type, magnification, intent of the experiment, conclusion you can draw). Be sure to include your team color as well.

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