User:Madeleine Y. Bee/Notebook/CHEM-572 2014S/2014/04/22: Difference between revisions

From OpenWetWare
Jump to navigationJump to search
No edit summary
(fix raw html notebook nav)
 
(4 intermediate revisions by one other user not shown)
Line 2: Line 2:
|-
|-
|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Cell Tests</span>
|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Cell Tests</span>
|style="background-color: #F2F2F2" align="center"|<html><img src="/images/9/94/Report.png" border="0" /></html> [[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>[[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}}
|style="background-color: #F2F2F2" align="center"|[[File:Report.png|frameless|link={{#sub:{{FULLPAGENAME}}|0|-11}}]][[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|[[File:Resultset_previous.png|frameless|link={{#lnpreventry:{{FULLPAGENAME}}}}]][[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}}
|-
|-
| colspan="2"|
| colspan="2"|
Line 11: Line 11:


===Test Procedures===
===Test Procedures===
====Type I Collagen Test (Detection and Quantification)=====
=====Type I Collagen Test (Detection and Quantification)=====
*[http://www.chondrex.com/products/mouse-type-i-collagen-detection-kit Company procedure ]
*[http://www.chondrex.com/products/mouse-type-i-collagen-detection-kit Company procedure ]


Line 23: Line 23:
=====Collagen Staining Procedure & Microscope Imaging Test=====
=====Collagen Staining Procedure & Microscope Imaging Test=====
*Absorbance data:
*Absorbance data:
**Original
[[Image:2014_0422_abs_collagen_quantification_test.PNG]]
**pD only
**pD & original
**Brute force
**Plain


*Calculations (from company procedure)
*Calculations (from company procedure)
Line 33: Line 29:
**Collagen (ug/3 scaffold layers)= [OD 540 value-(OD 605 value x 0.291)]/0.0378
**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
**Non-collagenous proteins (ug/3 scaffold layers)= OD 605 value/0.00204
***Original
[[Image:2014_0422_collagen_quantification_calcs.png]]
***pD only
***pD & original
***Brute force
***Plain
 
*Microscope images?


=====Trypsin Cell Detachment Procedure & Cell Counting=====
=====Trypsin Cell Detachment Procedure & Cell Counting=====
Line 51: Line 41:
===Next time===  
===Next time===  
*Perform all three tests in triplicate
*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


<!-- ##### DO NOT edit below this line unless you know what you are doing. ##### -->
<!-- ##### DO NOT edit below this line unless you know what you are doing. ##### -->

Latest revision as of 23:54, 26 September 2017

Cell Tests Main project page
Previous entry      Next entry

April 22, 2014

Objectives

  • Cell tests (only 2&3)

Test Procedures

Type I Collagen Test (Detection and Quantification)
Collagen Staining Procedure & Microscope Imaging
Trypsin Cell Detachment Procedure & Cell Counting

Test Results

Collagen Staining Procedure & Microscope Imaging Test
  • Absorbance data:

  • 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

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


Retrieved from "https://openwetware.org/mediawiki/index.php?title=User:Madeleine_Y._Bee/Notebook/CHEM-572_2014S/2014/04/22&oldid=1014454"