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Single-cell gene expression analysis technologies (Which one to select…
Single-cell gene expression analysis technologies
What for?
How
Cell isolation methods
FACS
Microfluidic circuits
Microdroplets
In situ
RNA detection
RNAseq
Reverse transcription
cDNA amplication
Library construction
Transcript converage
Barcoding
Quantification
RT-PCR
Multiplex
RT
Cell lysis
cDNA amplication
Quantification
FISH
Single-molecule FISH
High-throughput (cell)
Large-scale multiplex
encoding schemes
Labeling schemes
Signal amplification
Quantification
Spatial resolution
Which one to select
AHP
GOAL: SC gene expression analysis
CRITERION 1: Technical capabilities
Cellular throughput
Multiplex capability
Detection limit
Transcript coverage
Spatial information
Phenotypic information
Quantification
CRITERION 2: Logistics
Cell capture/recovery rate
Cell size limitations
Long-term cell storage
Processing time
CRITERION 3: Cost
Capital cost
Operating cost
Labour cost
CRITERION 3: Accessibility
Experimental protocol
Analysis
Setup
Open access for customization
Performance matrix
Case studies
Rare cell detection - HSC
Identification and characterization of cell populations