(Difference between revisions)
 Revision as of 14:21, 22 April 2014 (view source)← Previous diff Current revision (13:57, 23 April 2014) (view source) 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=====

## Current revision

Cell Tests Main project page
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## April 22, 2014

### Objectives

• Cell tests (only 2&3)

### 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