BioLT

Workflow

Cell Culture Workflow

Seeding, counting, and media handling checklist

This workflow connects daily maintenance, counting, and seeding decisions into one repeatable sequence. It is intended for routine mammalian cell culture work where consistency matters more than speed alone.

Steps

  1. Label plates, flasks, and waste containers before opening the hood.
  2. Warm complete medium and detachment reagents to the correct temperature.
  3. Harvest cells gently and resuspend until clumps are minimized.
  4. Count cells with a hemocytometer and confirm viability assumptions.
  5. Use the seeding calculator to match plate format, density, and final volume.
  6. Mix again immediately before dispensing to reduce settling bias.
  7. Record the actual seeded density, volume, passage number, and unusual observations.

Control points

  • Confirm that morphology and contamination status are acceptable before passage.
  • Check whether the intended density fits the plate surface area and experiment timeline.
  • Add overage for pipetting loss and dead volume instead of improvising at the bench.

Use these tools

  • Cell Seeding
  • Hemocytometer

Tips

  • Pre-fill edge wells when appropriate to reduce evaporation artifacts.
  • Do not rely on the initial suspension mix if the dispense takes several minutes.
  • Record viability and passage number because both can explain downstream variability.