A core skill in modern immunology is the ability to translate a biological question into the right experimental approach. 
 
𝗤𝘂𝗲𝘀𝘁𝗶𝗼𝗻 → 𝗠𝗲𝘁𝗵𝗼𝗱 → 𝗜𝗻𝘀𝗶𝗴𝗵𝘁 
 
𝗘𝘅𝗮𝗺𝗽𝗹𝗲𝘀: 
1. Which immune subpopulations drive therapy resistance? 
Method: Single-cell RNA-seq (Day 80) 
Insight: Reveals T cell exhausted subsets, suppressive myeloid populations, or unique transcriptional states influencing checkpoint therapy response. 
 
2. How does the tumor microenvironment regulate T cell activation? 
Method: Spatial Transcriptomics (Day 79) https://lnkd.in/eU6Du4ec 

Insight: Maps cytokine gradients, stromal barriers, and immune niches 𝘪𝘯 𝘴𝘪𝘵𝘶 . 
 
3. Which genes are essential for cytotoxicity or persistence? 
Method: CRISPR screens (Day 72) https://lnkd.in/eC_QZyR2 

Insight: Identifies regulators of exhaustion, memory formation, or CAR-T functionality. 
 
4. Can we model tumor–immune interactions 𝘦𝘹 𝘷𝘪𝘷𝘰? 
Method: Organoids and tumor-immune co-cultures (Day 76) https://lnkd.in/epxvJQQb 

Insight: Predicts responses to immunotherapies, studies infiltration, and models antigen escape. 
 
5. How do engineered receptors behave under controlled stimulation? 
Method: Optogenetics (Day 75) https://lnkd.in/e_pKFZGJ 

Insight: Light-controllable immune signaling reveals dose-response kinetics and circuit logic. 
 
𝗤𝘂𝗲𝘀𝘁𝗶𝗼𝗻 𝗳𝗼𝗿 𝘁𝗵𝗲 𝗔𝘂𝗱𝗶𝗲𝗻𝗰𝗲 
Which technique do you wish more immunologists understood how to use properly? 
 
Stay tuned for 𝗗𝗮𝘆 𝟴𝟯: Hot vs. 𝗖𝗼𝗹𝗱 𝗧𝘂𝗺𝗼𝗿𝘀 – 𝗶𝗺𝗺𝘂𝗻𝗲-𝗜𝗻𝗳𝗹𝗮𝗺𝗲𝗱 𝗮𝗻𝗱 𝗜𝗺𝗺𝘂𝗻𝗲-𝗘𝘅𝗰𝗹𝘂𝗱𝗲𝗱 𝗖𝗮𝗻𝗰𝗲𝗿𝘀 
 
#Immunology #TranslationalImmunology #SingleCell #SpatialTranscriptomics #CyTOF #CRISPR #ATACseq #Organoids #Microfluidics #Optogenetics #100DaysOfImmunology #SystemsImmunology