Fluorescence Upconversion
Gaussian-deconvolve gate width from a fluorescence-upconversion cross-correlation.
Description
Gaussian width deconvolution subtracts squared gate width from measured cross-correlation width.
Gaussian width deconvolution subtracts squared gate width from measured cross-correlation width. Width conventions must match.1
Gaussian-deconvolve gate width from a fluorescence-upconversion cross-correlation.
When to use Fluorescence Upconversion
Use this tool to gaussian-deconvolve gate width from a fluorescence-upconversion cross-correlation.
Prepare the optical measurements, geometry, fitted coefficients or caller-selected thresholds requested. Confirm wavelength, radius/diameter, pulse-width, angle and unit conventions before running the calculation.
How to use Fluorescence Upconversion and understand its results
- Prepare the optical measurements, geometry, fitted coefficients or caller-selected thresholds requested. Confirm wavelength, radius/diameter, pulse-width, angle and unit conventions before running the calculation.
- Enter measuredCrossCorrelationFemtoseconds, gatePulseWidthFemtoseconds using the field table and calculation rules, then run the tool.
- Read deconvolvedFluorescenceWidthFemtoseconds, resolvable. The explanation below defines the units, signs, ordering and any flags or placeholder values.
- If the tool reports an input issue, correct the values and run it again. Apply the stated assumptions when interpreting a successfully returned result.
| Input field | Meaning | Value type; see rules below | Requirement |
|---|---|---|---|
| measuredCrossCorrelationFemtoseconds | Measured upconversion width (fs) | number; minimum: 0 | Required |
| gatePulseWidthFemtoseconds | Gate pulse width (fs) | number; minimum: 0 | Required |
Enter nonnegative fs widths. Return deconvolved width and resolvable only when the squared difference is strictly positive. Equality or a smaller measured width returns zero with false.
deconvolved=sqrt(max(0,measured²−gate²)) Units, indexing, boundary conditions and exceptional cases are defined in the calculation rules above.
| Output field | Meaning | Value type |
|---|---|---|
| deconvolvedFluorescenceWidthFemtoseconds | Deconvolved fluorescence width (fs) | number; minimum: 0 |
| resolvable | Measured width exceeds gate width | boolean |
{
"input": {
"measuredCrossCorrelationFemtoseconds": 500,
"gatePulseWidthFemtoseconds": 300
},
"output": {
"deconvolvedFluorescenceWidthFemtoseconds": 400,
"resolvable": true
}
}Limitations and assumptions
No instrument-response fit or physical negative-width recovery is performed. Non-Gaussian shapes require another model.
Numeric inputs must be finite. Follow the calculation-specific rules for overflow, underflow and exceptional cases. Missing required fields, extra fields, wrong value types and violations of the calculation rules are input errors. A successfully returned false flag is an evaluated outcome; its meaning is explained above.
Alternative or Complementary methods
Use Transient-Absorption Lifetime Fit: Fit a positive transient-absorption trace to a single exponential using log-linear regression. For a complementary task, use FLIM Mean Lifetime: Calculate a count-weighted mean fluorescence lifetime from a supplied histogram.
References
-
Fluorescence Upconversion — tool.io
Similar or alternative tools
- Transient-Absorption Lifetime Fit
Fit a positive transient-absorption trace to a single exponential using log-linear regression.
- FLIM Mean Lifetime
Calculate a count-weighted mean fluorescence lifetime from a supplied histogram.