How is the object-space pixel size calculated?

It is calculated by dividing the physical camera pixel size (multiplied by the binning factor) by the total optical magnification (objective magnification multiplied by relay magnification).

What is the difference between optical magnification and screen magnification?

Optical magnification is the physical magnification of the lenses, whereas screen magnification depends on the display size and resolution when viewing the image.

How does the tool estimate optical resolution?

It uses the emission wavelength and numerical aperture (NA) to calculate the diffraction limits based on the Abbe and Rayleigh criteria.

What is the Nyquist sampling check?

It evaluates whether your pixel size is small enough (at least twice as small as the optical resolution limit) to resolve the finest details without aliasing.

Can I customize the appearance of the scale bar?

Yes, you can adjust the scale bar length, color, thickness, margin, position, and toggle the text label.

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Science & Education

Microscope Scale Bar Calibrator

Calculate microscope object scale, field of view, diffraction limits, Nyquist sampling, and download a transparent SVG scale bar.

Details

What this tool helps you do

Uses camera pixel pitch, binning, objective and relay magnification to calculate object-space microns per pixel. Optical resolution estimates use Abbe and Rayleigh criteria. The reported optical magnification is not screen viewing magnification.

Execution

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1 examples

40× fluorescence microscope calibration

Calibrates a 2048 × 1536 camera image with 3.45 µm pixels, a 40× objective, NA 1.0, and 550 nm emission.

<article><h2>0.08625 µm / pixel</h2><p>Field of view, diffraction limits, sampling assessment, and SVG overlay.</p></article>

Inputs

Options are grouped so long tools stay easier to scan.

14 options

SettingsAdjust formats, ranges, numbers, and modes.13

Image width (px)numberRequiredImage height (px)numberRequiredCamera pixel size (µm)numberRequiredObjective magnificationnumberRequiredRelay magnificationnumberRequiredCamera binningnumberRequiredEmission wavelength (nm)numberRequiredNumerical aperture (NA)numberRequiredScale bar length (µm, 0 = auto)numberOptionalScale bar colorcolorOptionalScale bar positionselectOptionalScale bar thickness (px)numberOptionalScale bar margin (px)numberOptional

TogglesEnable or disable optional behavior.1

Show labelcheckboxOptionalEnabled when checked

Result

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Samples

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imageWidthPx: 2048, imageHeightPx: 1536, cameraPixelSizeUm: 3.45, objectiveMagnification: 40, relayMagnification: 1, binning: 1, wavelengthNm: 509, numericalAperture: 1.3, barLengthUm: 20, barColor: '#ffffff', barPosition: 'bottom-right', barThicknessPx: 8, showLabel: true

Microscope Scale Bar Calibrator

What this tool helps you do

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Prepared example runs

Inputs

Result

Examples that match this tool

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Prepared example runs

Inputs

Result

Learn when to use this tool, what it supports, and how real users apply it.

Key facts

Overview

When to use

How it works

Use cases

Examples

1. Fluorescence Imaging Scale Bar Generation

2. Nyquist Sampling Verification for Super-Resolution

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