How is transfection efficiency calculated?

Efficiency is GFP-positive cells divided by total cells, multiplied by 100.

What does single measurement mode provide?

It provides the GFP-positive percentage and a Wilson 95% confidence interval.

What does replicate mode provide?

It calculates efficiency for each replicate, then reports the mean and sample standard deviation.

How should replicate values be entered?

Enter comma-separated totals and GFP-positive counts in the same order, with the same number of values.

Is this calculator suitable for clinical decisions?

No. It is intended for education and experimental planning, not clinical decisions.

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Transfection Efficiency Calculator (GFP Positive Rate)

GFP reporter transfection scoring: efficiency = GFP+ / total × 100 with a Wilson 95% CI in single mode and per-replicate mean ± SD in replicate mode. Interpretation bands from excellent (≥70%) to low (<10%) with optimization hints. Wilson 1927 / Newcombe 1998 / Kim & Eberwine 2010 derived. Educational use only.

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2 examples

Flow cytometry: 150 GFP+ of 500 cells → 30% transfection

A GFP-plasmid transfection read by flow cytometry counts 150 GFP-positive events among 500 viable single cells: efficiency = 150/500 = 30%, with a Wilson 95% confidence interval of roughly 26–34%. The tool classifies 30% as good — a typical result for many immortalized lines — and reminds that dead cells should be gated out before scoring.

{"result":"Transfection efficiency = 30% (150/500), 95% CI 26.15–34.16%.","metadata":{"input":{"mode":"single","totalCells":500,"gfpPositiveCells":150},"result":{"efficiencyPct":30,"ciLowerPct":26.15,"ciUpperPct":34.16}}}

Three microscopy fields: 25.0, 30.39, 25.74% → mean 27.04%

Three replicate fluorescence fields scored manually give 120/480, 155/510, and 130/505 GFP-positive cells — 25.00%, 30.39%, and 25.74%. The tool averages these to 27.04% with a sample SD of 2.92 percentage points and flags the spread between fields as a reminder to score several random fields per well rather than one cherry-picked image.

{"result":"Transfection efficiency = 27.04% ± 2.92 SD over 3 replicates.","metadata":{"input":{"mode":"replicates","replicateTotals":"480, 510, 505"},"result":{"replicateCount":3,"meanPct":27.04,"sdPct":2.92}}}

Inputs

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6 options

ContentPaste or type the main input values.2

Replicate Totals (comma-separated)textRequiredTotal cells per replicate well/field (replicate mode; same count as the GFP list).Replicate GFP Positives (comma-separated)textRequiredGFP-positive cells per replicate in the same order (replicate mode).

SettingsAdjust formats, ranges, numbers, and modes.4

Calculation ModeselectRequiredOne pooled count, or several replicate wells/fields for mean ± SD.Total Cells CountednumberRequiredTotal viable cells scored (single mode).GFP-Positive CellsnumberRequiredCells scoring above the GFP threshold (single mode).Decimal PlacesnumberOptional

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Transfection Efficiency Calculator (GFP Positive Rate)

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Examples

1. Flow cytometry: 150 GFP-positive cells out of 500

2. Three microscopy fields with replicate counts

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