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<p>Optical imaging is becoming a versatile tool in life science and biomedical research. Various conventional and advanced optical imaging techniques are used in the research. However, most biological samples are transparent; label-free images provide low contrast. Exogenous fluorophores can overcome this limitation by labelling it to the samples which will provide better contrast and additional morphological information. As we consider labelled imaging, fluorescent probes like organic dyes provide lower photostability and brightness along with a narrow emission spectrum. So as an alternative fluorescent probe, Quantum Dots (QDs) having wide emission spectra, and higher photostability are being used in bioimaging. For fluorescent probes like QDs, ligand-receptor pairs play a vital role in precise drug delivery, molecular tracking and cellular imaging. The current review describes the mechanism of ligand-receptor pairs inside the cellular components. Ligand-modified QDs target cell surface receptors for specific endocytosis uptake, through non-specific mechanisms. Internalized QDs localize to organelles via surface coatings, enabling bright, stable fluorescence for precise cancer cell tracking. Also, multivalent conjugation minimizes off-target effects, supports multiplexing, and amplifies signals for superior live imaging. Also, role of various QDs is explained which have been used to enhance the sensitivity and resolution of imaging techniques providing more specific molecular and cellular analysis by using different advanced optical imaging techniques such as Widefield microscopy, Confocal microscopy, Förster Resonance Energy Transfer (FRET), Fluorescence Lifetime Imaging Microscopy (FLIM), Fluorescence Recovery After Photobleaching (FRAP), Two-Photon Microscopy (TPM). As we consider TPM, it excels over confocal and widefield methods with more than 1&#xa0;mm penetration for thick samples, focal-plane excitation minimizing photodamage and photobleaching, higher signal-to-noise, and a single NIR wavelength for multicolor imaging. Considering the advantages among all imaging techniques, several studies suggested two-photon Microscopy (TPM) as very advantageous over other imaging techniques. This review summarises the revolutionising role of QDs and ligand-receptor pairs in optical imaging by referring to various studies and recent discoveries in advanced imaging techniques, offering innovative insights into biomedical research.</p>

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A review on fluorescent probes and ligand–receptor pairs for two-photon and multimodal optical microscopic techniques in cellular imaging

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2026年3月6日

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  • Varad Mahesh Phadke,
  • Shyamala Guruvare,
  • Anuradha Calicut Kini Rao,
  • Saritha Kamath U.,
  • Bharath Prasad A.S.,
  • Bhushan A. Kanagalekar,
  • Nirmal Mazumder,
  • Anand Krishna Asundi,
  • Ajeetkumar Patil

摘要

Optical imaging is becoming a versatile tool in life science and biomedical research. Various conventional and advanced optical imaging techniques are used in the research. However, most biological samples are transparent; label-free images provide low contrast. Exogenous fluorophores can overcome this limitation by labelling it to the samples which will provide better contrast and additional morphological information. As we consider labelled imaging, fluorescent probes like organic dyes provide lower photostability and brightness along with a narrow emission spectrum. So as an alternative fluorescent probe, Quantum Dots (QDs) having wide emission spectra, and higher photostability are being used in bioimaging. For fluorescent probes like QDs, ligand-receptor pairs play a vital role in precise drug delivery, molecular tracking and cellular imaging. The current review describes the mechanism of ligand-receptor pairs inside the cellular components. Ligand-modified QDs target cell surface receptors for specific endocytosis uptake, through non-specific mechanisms. Internalized QDs localize to organelles via surface coatings, enabling bright, stable fluorescence for precise cancer cell tracking. Also, multivalent conjugation minimizes off-target effects, supports multiplexing, and amplifies signals for superior live imaging. Also, role of various QDs is explained which have been used to enhance the sensitivity and resolution of imaging techniques providing more specific molecular and cellular analysis by using different advanced optical imaging techniques such as Widefield microscopy, Confocal microscopy, Förster Resonance Energy Transfer (FRET), Fluorescence Lifetime Imaging Microscopy (FLIM), Fluorescence Recovery After Photobleaching (FRAP), Two-Photon Microscopy (TPM). As we consider TPM, it excels over confocal and widefield methods with more than 1 mm penetration for thick samples, focal-plane excitation minimizing photodamage and photobleaching, higher signal-to-noise, and a single NIR wavelength for multicolor imaging. Considering the advantages among all imaging techniques, several studies suggested two-photon Microscopy (TPM) as very advantageous over other imaging techniques. This review summarises the revolutionising role of QDs and ligand-receptor pairs in optical imaging by referring to various studies and recent discoveries in advanced imaging techniques, offering innovative insights into biomedical research.