Description
Compare supplied before/after fluorescence levels relative to a background.
Biophysical scalar models require explicit signal, concentration, temperature, energy and geometric conventions. Effective concentration differs from dimensionless activity, a single displacement differs from mean-square displacement, and a fluorescence yield ratio needs absorption and refractive-index corrections. Supplied model parameters do not establish that the model applies to an experiment.
Compare supplied before/after fluorescence levels relative to a background.
When to use this tool
Use Fluorophore Bleach Step Height when you already have beforeLevel, afterLevel, backgroundLevel and need to compare supplied before/after fluorescence levels relative to a background.
How to use the tool and how it works
- Enter beforeLevel, afterLevel, backgroundLevel. Use the units and conventions below.
- Run the tool. Correct any format, range, or cross-field input error before interpreting the output.
- Read stepHeight, backgroundCorrectedBefore, relativeStep. Use the output convention below.
Require beforeLevel>backgroundLevel; step=before-after, correctedBefore=before-background, relativeStep=step/correctedBefore.1
| Input field | Meaning and units | Accepted input |
|---|---|---|
| beforeLevel | Before Level | Required. Finite number. |
| afterLevel | After Level | Required. Finite number. |
| backgroundLevel | Background Level | Required. Finite number. |
| Output field | Interpretation | Returned shape |
|---|---|---|
| stepHeight | Step Height | Required. Finite number. |
| backgroundCorrectedBefore | Background Corrected Before | Required. Finite number. |
| relativeStep | (beforeLevel-afterLevel)/(beforeLevel-backgroundLevel); can be negative or above one, without counting molecules. | Required. Finite number. |
{
"input": {
"beforeLevel": 120,
"afterLevel": 90,
"backgroundLevel": 20
},
"output": {
"stepHeight": 30,
"backgroundCorrectedBefore": 100,
"relativeStep": 0.3
}
}Limitations and assumptions
- After-level is unrestricted, so negative or above-one relative steps are accepted. No bleaching, fluorophore count, background or actual molecular event is inferred.
- Inputs must satisfy both the field types and the cross-field conditions above. Invalid input causes an error; a successful false flag or documented placeholder is a different result.
- Numeric calculations use binary floating point unless exact decimal-integer arithmetic is explicitly stated. No hidden measurement tolerance or decimal rounding is added; nonfinite or out-of-contract results are rejected. Numeric signed-zero results are exported as zero.
Alternative or Complementary methods
Use the complete physical model, calibrated measurements and uncertainty analysis. Check reference states, photon-signal bases, diffusion assumptions and binding-constant conventions before interpreting local contributions as physical measurements.
References
Similar or alternative tools
- Fluorescence Quantum Yield Reference Ratio
Calculate a supplied relative fluorescence quantum-yield term.
- Fluorophore Background Shot Noise Reference
Calculate a Poisson background-count standard-deviation reference.