What does the viability percentage measure?

It is the percentage of counted cells that are live: live cells ÷ (live cells + dead cells) × 100.

What dilution factor should I enter for a 1:1 trypan blue mix?

Enter 2 because the final dye mixture contains one part sample and one part trypan blue.

What does the squares count represent?

It is the number of 1 mm² large hemocytometer squares included in the cell counts.

How are live cells per mL calculated?

The calculation uses the live-cell count, dilution factor, 10⁴, and the number of large squares counted.

Is this calculator intended for clinical decisions?

No. It is for educational use and experiment planning, not clinical diagnosis or treatment decisions.

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Trypan Blue Cell Viability Calculator

Viability % and live/dead/total cells per mL from trypan blue exclusion counts, with culture-health interpretation. Educational use only.

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2 examples

180 live + 20 dead in 4 squares, 1:1 dye → 90% viability

A 1:1 trypan blue mix (dilution factor 2) counted over 4 large squares gives 180 bright and 20 blue cells. Viability is 180 ÷ 200 = 90%, with 9×10⁵ live cells/mL and 1×10⁶ total cells/mL — a healthy culture ready to passage.

{"result":"Viability ≈ 90% (180 live / 20 dead); live ≈ 900,000 cells/mL, total ≈ 1,000,000 cells/mL. Culture looks healthy.","metadata":{"result":{"viabilityPercent":90,"livePerMl":900000,"totalPerMl":1000000,"healthStatus":"healthy"}}}

55 live + 45 dead → 55% viability, stressed culture

An over-confluent flask counted neat (dilution 1, single square) shows 55 bright versus 45 blue cells. The 55% viability lands in the stressed band, and the tool flags overconfluency, harsh dissociation, and mycoplasma as likely causes to investigate before experiments.

{"result":"Viability ≈ 55% (55 live / 45 dead); live ≈ 550,000 cells/mL, dead ≈ 450,000 cells/mL. Culture is stressed.","metadata":{"result":{"viabilityPercent":55,"livePerMl":550000,"healthStatus":"stressed"}}}

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5 options

SettingsAdjust formats, ranges, numbers, and modes.5

Live (Unstained) CellsnumberRequiredBright, dye-excluding cells counted.Dead (Blue) CellsnumberRequiredDeep-blue, dye-permeable cells counted.Dilution FactornumberRequiredTotal-to-sample volume ratio of the dye mix (1:1 with trypan blue ⇒ 2).Squares CountednumberRequiredNumber of 1 mm² large squares the counts cover.Decimal PlacesnumberOptional

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Trypan Blue Cell Viability Calculator

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1. Healthy culture with a 1:1 trypan blue mix

2. Stressed culture after an over-confluent flask

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