User:Madeleine Y. Bee/Notebook/CHEM-572 2014S/2014/04/22: Difference between revisions
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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> | |style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Cell Tests</span> | ||
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== | ==April 22, 2014== | ||
* | ===Objectives=== | ||
*Cell tests (only 2&3) | |||
===Test Procedures=== | |||
=====Type I Collagen Test (Detection and Quantification)===== | |||
*[http://www.chondrex.com/products/mouse-type-i-collagen-detection-kit Company procedure ] | |||
=====Collagen Staining Procedure & Microscope Imaging===== | |||
*[http://www.chondrex.com/products/sirius-red-fast-green-collagen-staining-kit Company procedure] | |||
=====Trypsin Cell Detachment Procedure & Cell Counting===== | |||
*[http://www.sigmaaldrich.com/content/dam/sigma-aldrich/docs/Sigma/General_Information/1/celldetachmentprotocol.pdf Company procedure] | |||
===Test Results=== | |||
=====Collagen Staining Procedure & Microscope Imaging Test===== | |||
*Absorbance data: | |||
[[Image:2014_0422_abs_collagen_quantification_test.PNG]] | |||
*Calculations (from company procedure) | |||
**"To calculate the amount of collagen, correct the OD 540 value by subtracting the contribution of Fast Freen at 540nm, which is 29.1% of the OD 605 value. The color equivalence (OD values/ug protein) is 0.0378 for collagen and 0.00204 for non-collagenous protein at OD 540 and 605, respectively." | |||
**Collagen (ug/3 scaffold layers)= [OD 540 value-(OD 605 value x 0.291)]/0.0378 | |||
**Non-collagenous proteins (ug/3 scaffold layers)= OD 605 value/0.00204 | |||
[[Image:2014_0422_collagen_quantification_calcs.png]] | |||
=====Trypsin Cell Detachment Procedure & Cell Counting===== | |||
Performed on 3 scaffold layers. | |||
*“Original” - 125 cells on scaffold | |||
*Polydopmaine & “original” - 450 cells on scaffold | |||
*Polydopmaine only- 150 cells on scaffold | |||
*Brute force - 300 cells on scaffold | |||
*Plain - 100 cells on scaffold | |||
===Next time=== | |||
*Perform all three tests in triplicate | |||
*Have one scaffold for assays that has no cells one it (for blank & with "leftover" media [under microscope?]) | |||
*Keep solutions post-assay to look at under microscope to see if cells are still there | |||
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Revision as of 11:57, 23 April 2014
Cell Tests | <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page <html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html> </html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html> |
April 22, 2014Objectives
Test ProceduresType I Collagen Test (Detection and Quantification)Collagen Staining Procedure & Microscope ImagingTrypsin Cell Detachment Procedure & Cell CountingTest ResultsCollagen Staining Procedure & Microscope Imaging Test
Trypsin Cell Detachment Procedure & Cell CountingPerformed on 3 scaffold layers.
Next time
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