Lecture Notes - Tumor Immunology - week 3

Module 2: Tumor Immunology (Week 3)

LECTURE 21: TOOLS IN TUMOR IMMUNOLOGY/SINGLE CELL TECH.




Cytometry examines cell characteristics: size, count, morphology, phase/cycle, DNA content & presence of
specific protein  HD cytometry: addressing molecule expression at single cell level (highly specific); allows
discovery of rare cells/molecules

, Module 2: Tumor Immunology (Week 3)

Cytometry allows examination of cells as
single cell/individual: behavioral change
upon certain stimulation (e.g. oncogenic
mutation) & response of each cell to certain
drug




Fundamental principle of cytometry (conventional)


Determination of normal & abnormal
functions of the cells

Single cell suspension goes through the
sheath  laser dispersion (light source) 
collected by optics system (prism, mirror,
filter)  electronic amplification 
analysis




Examining the morphology & functional
characteristics from tissue biopsy

Cells extracted from tissue sample is loaded with
antigen  put through the suspension tube 
analysis of morphology & function at the same time
(TOF-MS)

Different wavelength/light source to examine
different function  composite/concomitant
analysis (e.g. cell status, immune cell types, regulatory
functions/Treg, IDO, etc, stromal conditions)

, Module 2: Tumor Immunology (Week 3)

HD Cytometry (less noise d/t reduced scatter)

Metal-elements based: mass cytometry (using rare natural elements)

Signal detection affected by: antigen level, sensitivity of sensor, background signals & cross-talk between
abundance sensitivity, isotope purity, and metal ion oxidation

, Module 2: Tumor Immunology (Week 3)




Fluorescence-based:

 BD Symphony (each light source emits 1 wavelength): risk of cross-talk & spillover >>> (non-specific)
 using PMT (photon multiplier tube)



PMT