Which microscope do you need to image a tissue section that is stained with FITC (fluorescein isothiocyanate)-conjugated antibody?

Question

BlackTom AI · Accepted Answer

Florophores such as FITC absorb blue light and emit green light, so imaging FITC requires an instrument that can excite the fluorophore and selectively collect its emitted fluorescence.

1. Relevant knowledge points and concepts
- FITC (fluorescein isothiocyanate) is a fluorescent dye with an excitation peak around ~490–495 nm (blue) and an emission peak around ~515–520 nm (green).
- A fluorescence microscope contains an excitation light source (e.g., mercury/xenon lamp or LED), excitation and emission filters, and a dichroic mirror to separate excitation and emitted light so the faint fluorescence signal can be observed or imaged.
- Other microscope types (brightfield light microscope, phase contrast, polarizing, electron) do not provide the excitation/emission filter/dichroic arrangement required to selectively detect fluorophore emission.

2. Step-by-step reasoning
- The tissue is stained with an FITC-conjugated antibody, so the signal of interest is FITC fluorescence, not simple transmitted light contrast or electron-density contrast.
- To detect FITC you must illuminate the sample with the appropriate excitation wavelength and collect the specific emission wavelength while blocking the excitation light; this is precisely what a fluorescence microscope is designed to do (filter cubes or tunable filter sets accomplish this).
- Therefore the correct instrument is a fluorescence microscope.

3. Calculations/intermediate steps
- No numerical calculations are needed; the selection depends on matching instrument capabilities to FITC’s spectral properties (excite ~495 nm, detect ~515 nm).

4. Why the correct answer is correct
- A fluorescence microscope provides the excitation source, filter sets, and optics to excite FITC and capture its emitted green fluorescence, yielding high-contrast images of antibody labeling.

5. Why other options are incorrect (brief)
- Electron microscope: detects electron scattering/absorption, incompatible with optical fluorescence labeling.
- Polarizing microscope: detects birefringence and polarization effects, not fluorescent emission.
- Light (brightfield) microscope: uses transmitted white light and cannot selectively detect faint fluorescence without fluorescence optics.
- Phase contrast microscope: enhances contrast for transparent specimens but does not provide fluorescence excitation/emission filtering.

6. Conclusion
- The correct choice is Fluorescence microscope, which matches the provided answer.

Which microscope do you need to image a tissue section that is stained with FITC (fluorescein isothiocyanate)-conjugated antibody?单项选择题

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