t cell study methods

what can happen to Tregs without AIRE

Expect

autoimmunity

and impaired

immune function

• Some population

of self-reactive

cells (so differciation)

how can scientists study treg dysfunction

using in vivo or in silico models with controlled MHC expression that regulates T cell development through genetic modificaiton of mouse models (GEMMs)

after collection of mouse models, what is the next step in analysis

Mechanical and enzymatic (such

as collagenase) digestion break

down matrix and intercellular

connections

• Without single cell suspensions,

flow cytometry analysis and

single cell sequencing (i.e. TCR)

is confounded

how does magnetic bead isolate help in single cell population obtained

abelled magnetic beads for specific targets (i.e. CD45) can be used to

enrich/deplete labelled populations to improve yield following flow sorting

contrast to magnetic bead isolation, how can you obtain multi-labeled specific cell pop.

Flow:

Cells can be labelled and enriched (i.e. magnetic beads) before fluorescent antibody labelling and sorting

After desired cell populations are prepared, histograms or dot/scatter plots can

be used to analyze presence of desired positive or negative labelled populations

• Setting desired “gates” allows for analysis or sorting of sought-after cell type

• Higher abundance is indicated by moving right and up on the dot plo

tcr seuencing + next generation sequencing