Addressing the autofluorescence issue in deep tissue imaging by two-photon microscopy: the significance of far-red emitting dyes

Jun, Yong Woong; Kim, Hye Rim; Reo, Ye Jin; Dai, Mingchong; Ahn, Kyo Han

doi:10.1039/c7sc03362a

Cited by 143 publications

(136 citation statements)

References 40 publications

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“…As mentioned above, TPFM overcomes autofluorescence from intrinsic biomolecules using TPA-active dyes with emission maximaa bove 625 nm. [4] It is knownt hat planarization and p-expansion of ac hromophore often resultsi nt he bathochromic shift of absorption and emission maxima. Our recent reports have shown an elegant approach for the expansiono f the diketopyrrolopyrrole p-system, based on ad ouble intramolecular Friedel-Crafts reaction.…”

Section: Design and Synthesismentioning

confidence: 99%

“…Secondly, TPFM overcomes autofluorescence issues intrinsic to biomolecules due to the long excitation wavelengths. Thirdly, it grants an unprecedented imaging depth in the order of hundreds of micrometers whilst allowing three‐dimensional resolution at the submicrometer level.…”

Section: Introductionmentioning

confidence: 99%

“…[3] Firstly,i tu ses non-phototoxicn ear-infrared (NIR) irradiation that dramatically reduces photobleaching and photodynamic tissue damage,a llowing for the imaging of living specimens.S econdly,T PFM overcomes autofluorescence issues intrinsict ob iomolecules due to the long excitation wavelengths. Thirdly, it grants an unprecedentedi maging depth in the order of hundredso fm icrometers [4] whilst allowing threedimensional resolutiona tt he submicrometerlevel.…”

Section: Introductionmentioning

confidence: 99%

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Modified Isoindolediones as Bright Fluorescent Probes for Cell and Tissue Imaging

Vakuliuk

Jun

Vygranenko

et al. 2019

Chemistry A European J

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New L‐shaped fluorophores possessing five conjugated rings have been synthesized through a four‐step procedure involving diketopyrrolopyrrole synthesis and its double N‐alkylation, followed by trimethylsilyl bromide‐mediated rearrangement to thieno[2,3‐f]isoindole‐5,8‐dione and an intramolecular Friedel–Crafts reaction. In comparison with the parent isoindolediones and π‐expanded diketopyrrolopyrroles, these new dyes show red‐shifted absorption and emission (up to ≈630 nm). Their structural rigidity is responsible for both the observed small Stokes shifts and large fluorescence quantum yields. Tissue imaging studies revealed that these new dyes show advantageous features including minimal autofluorescence interference and pronounced solvent‐sensitive emission. Interestingly, there is a fundamental difference between a dye possessing an amino group and its analog bearing an N‐alkyl substituent. The former dye under two‐photon excitation at 900 nm gives bright images whereas its N‐alkylated counterpart does not. A new type of membrane localization has been discovered by an N‐alkylated isoindoledione possessing a benzofuryl substituent. In spite of the fact that the fluorescence quantum yield of this dye in a range of solvents is rather low, it does stain cell membranes exclusively. This new mode of cellular staining opens the door towards further development of membrane staining dyes.

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Section: Design and Synthesismentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Modified Isoindolediones as Bright Fluorescent Probes for Cell and Tissue Imaging

Vakuliuk

Jun

Vygranenko

et al. 2019

Chemistry A European J

Self Cite

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“…AF can be attributed to biomolecules such as NADH, folic acid, and retinol which have emission maxima in the range of 450-500 nm, and riboflavin, flavin coenzymes, and flavoproteins which have emission maxima in the range of 520-540 nm. 18 . AF creates a background that can impair the detection of dim markers emitting light at the same wavelengths In spectral flow cytometry, the spectral profile of unstained cells can be collected and treated as an independent parameter (AF), which allows the AF signature to be extracted using the unmixing algorithm.…”

Section: D) Autofluorescence (Af)mentioning

confidence: 99%

Design and Optimization Protocol for High-Dimensional Immunophenotyping Assays using Spectral Flow Cytometry

Ferrer‐Font

Pellefigues

Mayer

et al. 2019

Preprint

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Technological advances in fluorescence flow cytometry and an ever-expanding understanding of the complexity of the immune system has led to the development of large 20+ flow cytometry panels. Yet, as panel complexity and size increases, so does the difficulty involved in designing a high-quality panel, accessing the instrumentation capable of accommodating large numbers of parameters, and in analysing such high-dimensional data.A recent advancement is spectral flow cytometry, which in contrast to conventional flow cytometry distinguishes the full emission spectrum of each fluorochrome across all lasers, rather than identifying only the peak of emission. Fluorochromes with a similar emission maximum but distinct off-peak signatures can therefore be accommodated within the same flow cytometry panel, allowing greater flexibility in terms of panel design and fluorophore detection.Here, we highlight the specific characteristics regarding spectral flow cytometry and aim to guide users through the process of building, designing and optimising high-dimensional spectral flow cytometry panels using a comprehensive step-by-step protocol. Special considerations are also given for using highlyoverlapping dyes and a logical selection process an optimal marker-fluorophore assignment is provided.

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“…Finally,weevaluated the ability of DMONs-PEI to deliver mCherry mRNA( with the same length to EGFP mRNA) in vivo in BALB/c mice to avoid the autofluorescence interference. [20] Strong mCherry signal was detected after 48 hi na xillary,i nguinal and mesenteric lymph nodes for DMONs-PEI-mCherry mRNAt reated mice,w hile in vivo-jetPEI group treated mice exhibited fluorescence signal mainly at injection site and limited in lymph nodes (Figure 4A). Thea verage radiant efficiencya ts ignal sites was calculated and plotted in Figure 4B,w here DMONs-PEI exhibited comparable radiant efficiency to in vivo-jetPEI.…”

mentioning

confidence: 97%

Functional Nanoparticles with a Reducible Tetrasulfide Motif to Upregulate mRNA Translation and Enhance Transfection in Hard‐to‐Transfect Cells

et al. 2020

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Effective messenger RNA( mRNA) transfection in hard-to-transfect cells delivered by vectors is al ong-standing challenge.N ow it is hypothesizedt hat the high intracellular glutathione level is associated with suppressed mRNAt ranslation. This theory leads to an ew design principle of nextgeneration mRNAv ectors:n anoparticles with glutathione depletion chemistry upregulate mRNAt ranslation and enhance transfection, which is beneficial for mRNAd elivery in hard-to-transfect cells in vitro and in vivo.

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Addressing the autofluorescence issue in deep tissue imaging by two-photon microscopy: the significance of far-red emitting dyes

Abstract: We have developed a new class of two-photon absorbing dyes that are far-red emitting, water-soluble, and very bright inside cells as well as in tissue. The significant autofluorescence from yellow wavelength region in tissue imaging can be addressed by deep-red emitting dyes.

Cited by 143 publications

References 40 publications

Modified Isoindolediones as Bright Fluorescent Probes for Cell and Tissue Imaging

Modified Isoindolediones as Bright Fluorescent Probes for Cell and Tissue Imaging

Design and Optimization Protocol for High-Dimensional Immunophenotyping Assays using Spectral Flow Cytometry

Functional Nanoparticles with a Reducible Tetrasulfide Motif to Upregulate mRNA Translation and Enhance Transfection in Hard‐to‐Transfect Cells

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