characterizing cells
Download
Skip this Video
Download Presentation
Characterizing Cells

Loading in 2 Seconds...

play fullscreen
1 / 18

Characterizing Cells - PowerPoint PPT Presentation


  • 82 Views
  • Uploaded on

Characterizing Cells. What To Characterize. Confirmation Of Species Of Origin Correlation With The Tissue Of Origin Transformation Status Finite Or Continuous Cross-Contamination Stability (ex. susceptibility to transformation). How Is Characterization Done. Species Identification

loader
I am the owner, or an agent authorized to act on behalf of the owner, of the copyrighted work described.
capcha
Download Presentation

PowerPoint Slideshow about ' Characterizing Cells' - juliana-ortiz


An Image/Link below is provided (as is) to download presentation

Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author.While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server.


- - - - - - - - - - - - - - - - - - - - - - - - - - E N D - - - - - - - - - - - - - - - - - - - - - - - - - -
Presentation Transcript
what to characterize
What To Characterize
  • Confirmation Of Species Of Origin
  • Correlation With The Tissue Of Origin
  • Transformation Status
  • Finite Or Continuous
  • Cross-Contamination
  • Stability (ex. susceptibility to transformation)
slide3

How Is Characterization Done

  • Species Identification
    • Chromosomal analysis
  • Tissue Markers
    • Cell surface markers
      • Ex. CD11c if DC
    • Intermediate filament proteins
      • Ex. 1 Glial fibrillary acidic protein (GFAP) for astrocytes. Ex. 2 Desmin for muscle cells. Ex. 3 Cytokeratin for epithelial cells
    • Differentiated products
      • Ex. Melanin for melanocytes, hemoglogin for erythroid cells, serum albumin for hepatocytes
    • Unique Markers
      • Ex. HLA highly polymorphic, unique to an individual
  • Morphology
    • Easy and fast but has variability depending on culturing conditions and site
flow cytometry is a powerful technique for characterizing cells
Flow Cytometry Is A Powerful Technique For Characterizing Cells
  • Allows For Detection Of Surface Markers Of Cells
  • Allows For Detection Of Intracellular Factors
  • Allows Detection Of Secreted Factors By Cells
  • Allows For Detection Of DNA Content
how is it done
How Is It Done:

http://biology.berkeley.edu/crl/flow_cytometry_basic.html

limitations with light scattering
Limitations With Light Scattering
  • Some Information Can Be Obtained
  • FSC Correlates With Cell Size
  • SSC Correlates With Internal Complexity
  • To Distinguish Between 2 Cell types
    • A. Size Has To Be Different OR
    • B. Internal Complexity i.e amount of granules
  • If These Two Parameters Are The Same, Then No Distinction Can Be Made
  • See The Following Figure
fluorescent dyes and antibodies
Fluorescent Dyes And Antibodies
  • Fluorochromes Are Molecules That Emit Fluorescence Upon Excitation With Light
    • Ex. FITC (Fluorescein Isothiocyanate)
    • PE (Phycoerythrin)
    • PerCP (Peridinin Chlorophyll Protein)
    • APC (Allophycocyanin)
  • Some Fluorochromes Are Proteins, Some Are Small Organic Compounds
    • Ex. PE (Phycoerythrin)-Protein
    • Ex. FITC (Fluorescein Isothiocyanate)
principles of fluorescence
Principles Of Fluorescence

E= h f

 = f 

slide16

GM-CSF+IL-4

GM-CSF+IL-4+PGE2

Fig. 2

GMF: 1.85

GMF: 168.92

25

75

99

1

GMF: 11.82

GMF: 391.83

99

80

1

20

slide17

Unstained

ISOTYPES

ad