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Cell Transfection Principles and Complete Protocol

🚨 Failure Case Library (6) + Submit your own case

critical
Massive cell death after transfection — reagent toxicity is too high
Cells become rounded and detached after transfection; CCK-8 shows a sharp drop in viability vs. mock-transfected control.
💡 4 · ✓ 4
critical
Almost no fluorescence signal — transfection essentially failed
Under the fluorescence microscope, no GFP signal is visible; Western blot also shows essentially no target protein expression.
💡 5 · ✓ 5
severe
Very few cells are fluorescent — transfection efficiency is extremely low
Microscopy shows scattered positive cells; flow cytometry confirms positivity rate is very low (< 5 %).
💡 5 · ✓ 5
moderate
Very high replicate-well variation (poor reproducibility)
Parallel wells show very different fluorescence intensity / positive-cell rates; data fluctuates wildly between replicates.
💡 4 · ✓ 4
moderate
Co-transfection ratio imbalance — uneven dual-plasmid expression
Of two co-transfected plasmids, only one expresses strongly; the fraction of double-positive cells is low.
💡 5 · ✓ 4
minor
Granular Precipitate on Cells
Small granular precipitate visible microscopically on cells after adding transfection complexes
💡 3 · ✓ 4
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