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iFluor® 790 maleimide

HL-60 cells were incubated with (Red, +) or without (Green, -) Anti-human HLA-ABC (W6/32 mAb), followed by iFluor® 790 goat anti-mouse IgG conjugate. The fluorescence signal was monitored using ACEA NovoCyte flow cytometer in APC-Cy7 channel.
HL-60 cells were incubated with (Red, +) or without (Green, -) Anti-human HLA-ABC (W6/32 mAb), followed by iFluor® 790 goat anti-mouse IgG conjugate. The fluorescence signal was monitored using ACEA NovoCyte flow cytometer in APC-Cy7 channel.
Ordering information
Price ()
Catalog Number1366
Unit Size
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Additional ordering information
Telephone1-408-733-1055
Fax1-408-733-1304
Emailsales@aatbio.com
InternationalSee distributors
ShippingStandard overnight for United States, inquire for international
Physical properties
Molecular weight1439.86
SolventDMSO
Spectral properties
Correction Factor (260 nm)0.1
Correction Factor (280 nm)0.09
Extinction coefficient (cm -1 M -1)2500001
Excitation (nm)787
Emission (nm)812
Quantum yield0.131
Storage, safety and handling
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
StorageFreeze (< -15 °C); Minimize light exposure
UNSPSC12171501
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iFluor® 700 goat anti-mouse IgG (H+L)
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iFluor® 350 goat anti-mouse IgG (H+L) *Cross Adsorbed*
iFluor® 405 goat anti-mouse IgG (H+L) *Cross Adsorbed*
iFluor® 488 goat anti-mouse IgG (H+L) *Cross Adsorbed*
iFluor® 514 goat anti-mouse IgG (H+L) *Cross Adsorbed*
iFluor® 532 goat anti-mouse IgG (H+L) *Cross Adsorbed*
iFluor® 555 goat anti-mouse IgG (H+L) *Cross Adsorbed*
iFluor® 594 goat anti-mouse IgG (H+L) *Cross Adsorbed*
iFluor® 633 goat anti-mouse IgG (H+L) *Cross Adsorbed*
iFluor® 647 goat anti-mouse IgG (H+L) *Cross Adsorbed*
iFluor® 680 goat anti-mouse IgG (H+L) *Cross Adsorbed*
iFluor® 700 goat anti-mouse IgG (H+L) *Cross Adsorbed*
iFluor® 750 goat anti-mouse IgG (H+L) *Cross Adsorbed*
iFluor® 790 goat anti-mouse IgG (H+L) *Cross Adsorbed*
iFluor® 350 goat anti-rabbit IgG (H+L)
iFluor® 405 goat anti-rabbit IgG (H+L)
iFluor® 488 goat anti-rabbit IgG (H+L)
iFluor® 514 goat anti-rabbit IgG (H+L)
iFluor® 532 goat anti-rabbit IgG (H+L)
iFluor® 555 goat anti-rabbit IgG (H+L)
iFluor® 594 goat anti-rabbit IgG (H+L)
iFluor® 633 goat anti-rabbit IgG (H+L)
iFluor® 647 goat anti-rabbit IgG (H+L)
iFluor® 680 goat anti-rabbit IgG (H+L)
iFluor® 700 goat anti-rabbit IgG (H+L)
iFluor® 750 goat anti-rabbit IgG (H+L)
iFluor® 790 goat anti-rabbit IgG (H+L)
iFluor® 350 goat anti-rabbit IgG (H+L) *Cross Adsorbed*
iFluor® 405 goat anti-rabbit IgG (H+L) *Cross Adsorbed*
iFluor® 488 goat anti-rabbit IgG (H+L) *Cross Adsorbed*
iFluor® 514 goat anti-rabbit IgG (H+L) *Cross Adsorbed*
iFluor® 532 goat anti-rabbit IgG (H+L) *Cross Adsorbed*
iFluor® 555 goat anti-rabbit IgG (H+L) *Cross Adsorbed*
iFluor® 594 goat anti-rabbit IgG (H+L) *Cross Adsorbed*
iFluor® 633 goat anti-rabbit IgG (H+L) *Cross Adsorbed*
iFluor® 647 goat anti-rabbit IgG (H+L) *Cross Adsorbed*
iFluor® 680 goat anti-rabbit IgG (H+L) *Cross Adsorbed*
iFluor® 700 goat anti-rabbit IgG (H+L) *Cross Adsorbed*
iFluor® 750 goat anti-rabbit IgG (H+L) *Cross Adsorbed*
iFluor® 790 goat anti-rabbit IgG (H+L) *Cross Adsorbed*
iFluor® 350-streptavidin conjugate
iFluor® 405-streptavidin conjugate
iFluor® 488-streptavidin conjugate
iFluor® 514-streptavidin conjugate
iFluor® 532-streptavidin conjugate
iFluor® 555-streptavidin conjugate
iFluor® 594-streptavidin conjugate
iFluor® 633-streptavidin conjugate
iFluor® 647-streptavidin conjugate
iFluor® 680-streptavidin conjugate
iFluor® 700-streptavidin conjugate
iFluor® 750-streptavidin conjugate
iFluor 750™ RGD Conjugate
iFluor® 488 azide
iFluor® 405 hydrazide
iFluor® 800 acid
iFluor® 810 acid
iFluor® 820 acid
iFluor® 860 acid
iFluor® 546 goat anti-mouse IgG (H+L)
iFluor® 546 goat anti-mouse IgG (H+L) *Cross Adsorbed*
iFluor® 568 goat anti-mouse IgG (H+L)
iFluor® 568 goat anti-mouse IgG (H+L) *Cross Adsorbed*
iFluor® 546 goat anti-rabbit IgG (H+L)
iFluor® 546 goat anti-rabbit IgG (H+L) *Cross Adsorbed*
iFluor® 568 goat anti-rabbit IgG (H+L)
iFluor® 568 goat anti-rabbit IgG (H+L) *Cross Adsorbed*
iFluor® 555 alkyne
iFluor® 555 azide
iFluor® 546-streptavidin conjugate
iFluor® 568-streptavidin conjugate
iFluor® 488 alkyne
iFluor® 350 Styramide *Superior Replacement for Alexa Fluor 350 tyramide*
iFluor® 488 Styramide *Superior Replacement for Alexa Fluor 488 tyramide and Opal 520*
iFluor® 546 Styramide *Superior Replacement for Alexa Fluor 546 tyramide*
iFluor® 555 Styramide *Superior Replacement for Alexa Fluor 555 tyramide and Opal 570*
iFluor® 568 Styramide *Superior Replacement for Alexa Fluor 568 tyramide*
iFluor® 594 Styramide *Superior Replacement for Alexa Fluor 594 tyramide*
iFluor® 647 Styramide *Superior Replacement for Alexa Fluor 647 tyramide*
iFluor® 680 Styramide *Superior Replacement for Alexa Fluor 680 tyramide and Opal 690*
iFluor® 700 Styramide *Superior Replacement for Alexa Fluor 700 tyramide*
iFluor® 750 Styramide *Superior Replacement for Alexa Fluor 750 tyramide*
iFluor® 555 Tyramide
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iFluor® 350 PSA™ Imaging Kit with Goat Anti-Rabbit IgG
iFluor® 488 PSA™ Imaging Kit with Goat Anti-Rabbit IgG
iFluor® 555 PSA™ Imaging Kit with Goat Anti-Rabbit IgG
iFluor® 594 PSA™ Imaging Kit with Goat Anti-Rabbit IgG
iFluor® 647 PSA™ Imaging Kit with Goat Anti-Rabbit IgG
iFluor 350™ PSA™ Imaging Kit with Goat Anti-Mouse IgG
iFluor 488™ PSA™ Imaging Kit with Goat Anti-Mouse IgG
iFluor 555™ PSA™ Imaging Kit with Goat Anti-Mouse IgG
iFluor 594™ PSA™ Imaging Kit with Goat Anti-Mouse IgG
iFluor 647™ PSA™ Imaging Kit with Goat Anti-Mouse IgG
iFluor® 840 acid
iFluor® 790-streptavidin conjugate
iFluor® 800-streptavidin conjugate
iFluor® 820-streptavidin conjugate
iFluor® 840-streptavidin conjugate
iFluor® 860-streptavidin conjugate
iFluor® 488-Wheat Germ Agglutinin (WGA) Conjugate
iFluor® 555-Wheat Germ Agglutinin (WGA) Conjugate
iFluor® 594-Wheat Germ Agglutinin (WGA) Conjugate
iFluor® 647-Wheat Germ Agglutinin (WGA) Conjugate
iFluor® 488-Concanavalin A Conjugate
iFluor® 555-Concanavalin A Conjugate
iFluor® 594-Concanavalin A Conjugate
iFluor® 647-Concanavalin A Conjugate
iFluor® 350 Tyramide
iFluor® 546 Tyramide
iFluor® 568 Tyramide
iFluor® 594 Tyramide
iFluor® 488 TCO
iFluor® 555 TCO
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iFluor® 555 Tetrazine
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iFluor® 440-dUTP *1 mM in Tris Buffer (pH 7.5)*
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iFluor® 514 Styramide *Superior Replacement for Opal 540*
iFluor® 532 Styramide
iFluor® 633 Styramide *Superior Replacement for Opal 650*
iFluor® 440 Styramide
iFluor® 460 Styramide
iFluor® 610 Styramide
iFluor® 660 Styramide
iFluor® 405 Styramide
iFluor®488-PEG12-dUTP *1 mM in Tris Buffer (pH 7.5)*
iFluor®555-PEG12-dUTP *1 mM in Tris Buffer (pH 7.5)*
iFluor®647-PEG12-dUTP *1 mM in Tris Buffer (pH 7.5)*
iFluor® 800 goat anti-mouse IgG (H+L)
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iFluor® 810 goat anti-mouse IgG (H+L)
iFluor® 810 goat anti-mouse IgG (H+L) *Cross Adsorbed*
iFluor® 820 goat anti-mouse IgG (H+L)
iFluor® 820 goat anti-mouse IgG (H+L) *Cross Adsorbed*
iFluor® 840 goat anti-mouse IgG (H+L)
iFluor® 840 goat anti-mouse IgG (H+L) *Cross Adsorbed*
iFluor® 860 goat anti-mouse IgG (H+L)
iFluor® 860 goat anti-mouse IgG (H+L) *Cross Adsorbed*
iFluor® 800 goat anti-rabbit IgG (H+L)
iFluor® 800 goat anti-rabbit IgG (H+L) *Cross Adsorbed*
iFluor® 810 goat anti-rabbit IgG (H+L)
iFluor® 810 goat anti-rabbit IgG (H+L) *Cross Adsorbed*
iFluor® 820 goat anti-rabbit IgG (H+L)
iFluor® 820 goat anti-rabbit IgG (H+L) *Cross Adsorbed*
iFluor® 840 goat anti-rabbit IgG (H+L)
iFluor® 840 goat anti-rabbit IgG (H+L) *Cross Adsorbed*
iFluor® 860 goat anti-rabbit IgG (H+L)
iFluor® 860 goat anti-rabbit IgG (H+L) *Cross Adsorbed*
iFluor® 430 Tyramide *Superior Replacement for Opal 480*
iFluor® 450 Tyramide *Superior Replacement for Opal 480*
iFluor® 350 succinimidyl ester
iFluor® 405 succinimidyl ester
iFluor® 488 succinimidyl ester
iFluor® 514 succinimidyl ester
iFluor® 532 succinimidyl ester
iFluor® 555 succinimidyl ester
iFluor® 594 succinimidyl ester
iFluor® 633 succinimidyl ester
iFluor® 647 succinimidyl ester
iFluor® 660 succinimidyl ester
iFluor® 680 succinimidyl ester
iFluor® 700 succinimidyl ester
iFluor® 750 succinimidyl ester
iFluor® 610 succinimidyl ester
iFluor® 710 succinimidyl ester
iFluor® 800 succinimidyl ester
iFluor® 810 succinimidyl ester
iFluor® 820 succinimidyl ester
iFluor® 860 succinimidyl ester
iFluor® 546 succinimidyl ester
iFluor® 568 succinimidyl ester
iFluor® 430 succinimidyl ester
iFluor® 450 succinimidyl ester
iFluor® 840 succinimidyl ester
iFluor® 560 succinimidyl ester
iFluor® 670 succinimidyl ester
iFluor® 460 succinimidyl ester
iFluor® 440 succinimidyl ester
iFluor® 665 succinimidyl ester
iFluor® 690 succinimidyl ester
iFluor® Ultra 594 succinimidyl ester
iFluor® Ultra 647 succinimidyl ester
iFluor® Ultra 750 succinimidyl ester
iFluor® 720 succinimidyl ester
iFluor® 740 succinimidyl ester
iFluor® 597 succinimidyl ester
iFluor® 770 succinimidyl ester
iFluor® 780 succinimidyl ester
iFluor® 570 succinimidyl ester
iFluor® 830 acid
iFluor® 830 succinimidyl ester
iFluor™ 405 azide
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OverviewpdfSDSpdfProtocol


Molecular weight
1439.86
Correction Factor (260 nm)
0.1
Correction Factor (280 nm)
0.09
Extinction coefficient (cm -1 M -1)
2500001
Excitation (nm)
787
Emission (nm)
812
Quantum yield
0.131
In vivo fluorescence imaging uses a sensitive camera to detect the fluorescence emission from fluorophores in whole-body living small animals. To overcome the photon attenuation in living tissue, fluorophores with long emission at the near-infrared (NIR) region are generally preferred, including widely used small indocarbocyanine dyes. Recent advances in imaging strategies and reporter techniques for in vivo fluorescence imaging include novel approaches to improve the specificity and affinity of the probes and to modulate and amplify the signal at target sites for enhanced sensitivity. Further emerging developments aim to achieve high-resolution, multimodality, and lifetime-based in vivo fluorescence imaging. Our iFluor® 790 is designed to label proteins and other biomolecules with near-infrared fluorescence. Conjugates prepared with iFluor® 790 have excitation and emission spectra similar to that of indocyanine green (ICG) and the IRDye® 800, with 787/812 nm excitation/emission maxima. iFluor® 790 dye emission is well separated from commonly used far-red fluorophores such as Cy5, Cy7, or allophycocyanin (APC), facilitating multicolor analysis. This fluorophore is also useful for small animal in-vivo imaging applications or other imaging applications requiring NIR detections, such as the two-color western applications with the LI-COR® Odyssey® infrared imaging system.

Example protocol


PREPARATION OF STOCK SOLUTIONS

Unless otherwise noted, all unused stock solutions should be divided into single-use aliquots and stored at -20 °C after preparation. Avoid repeated freeze-thaw cycles.

1. iFluor™ 790 maleimide stock solution (Solution B)
Add anhydrous DMSO into the vial of iFluor™ 790 maleimide to make a 10 mM stock solution. Mix well by pipetting or vortex.
Note     Prepare the dye stock solution (Solution B) before starting the conjugation. Use promptly. Extended storage of the dye stock solution may reduce the dye activity. Solution B can be stored in freezer for upto 4 weeks when kept from light and moisture. Avoid freeze-thaw cycles.


2. Protein stock solution (Solution A)
Mix 100 µL of a reaction buffer (e.g., 100 mM MES buffer with pH ~6.0) with 900 µL of the target protein solution (e.g. antibody, protein concentration >2 mg/mL if possible) to give 1 mL protein labeling stock solution.
Note     The pH of the protein solution (Solution A) should be 6.5 ± 0.5.
Note     Impure antibodies or antibodies stabilized with bovine serum albumin (BSA) or other proteins will not be labeled well.
Note     The conjugation efficiency is significantly reduced if the protein concentration is less than 2 mg/mL. For optimal labeling efficiency the final protein concentration range of 2-10 mg/mL is recommended.

Optional: if your protein does not contain a free cysteine, you must treat your protein with DTT or TCEP to generate a thiol group. DTT or TCEP are used for converting a disulfide bond to two free thiol groups. If DTT is used you must remove free DTT by dialysis or gel filtration before conjugating a dye maleimide to your protein. Following is a sample protocol for generating a free thiol group:
  1. Prepare a fresh solution of 1 M DTT (15.4 mg/100 µL) in distilled water.
  2. Make IgG solution in 20 mM DTT: add 20 µL of DTT stock per ml of IgG solution while mixing. Let stand at room temp for 30 minutes without additional mixing (to minimize reoxidation of cysteines to cystines).
  3. Pass the reduced IgG over a filtration column pre-equilibrated with "Exchange Buffer". Collect 0.25 mL fractions off the column.
  4. Determine the protein concentrations and pool the fractions with the majority of the IgG. This can be done either spectrophotometrically or colorimetrically.
  5. Carry out the conjugation as soon as possible after this step (see Sample Experiment Protocol).
    Note     IgG solutions should be >4 mg/mL for the best results. The antibody should be concentrated if less than 2 mg/mL. Include an extra 10% for losses on the buffer exchange column.
    Note     The reduction can be carried out in almost any buffers from pH 7-7.5, e.g., MES, phosphate or TRIS buffers.
    Note     Steps 3 and 4 can be replaced by dialysis. 

SAMPLE EXPERIMENTAL PROTOCOL

This labeling protocol was developed for the conjugate of Goat anti-mouse IgG with iFluor™ 790 maleimide. You might need further optimization for your particular proteins.
Note     Each protein requires distinct dye/protein ratio, which also depends on the properties of dyes. Over labeling of a protein could detrimentally affects its binding affinity while the protein conjugates of low dye/protein ratio gives reduced sensitivity.


Run conjugation reaction
  1. Use 10:1 molar ratio of Solution B (dye)/Solution A (protein) as the starting point:  Add 5 µL of the dye stock solution (Solution B, assuming the dye stock solution is 10 mM) into the vial of the protein solution (95 µL of Solution A) with effective shaking. The concentration of the protein is ~0.05 mM assuming the protein concentration is 10 mg/mL and the molecular weight of the protein is ~200KD.
    Note     We recommend to use 10:1 molar ratio of Solution B (dye)/Solution A (protein). If it is too less or too high, determine the optimal dye/protein ratio at 5:1, 15:1 and 20:1 respectively.
  2. Continue to rotate or shake the reaction mixture at room temperature for 30-60 minutes. 

Purify the conjugation
The following protocol is an example of dye-protein conjugate purification by using a Sephadex G-25 column.
  1. Prepare Sephadex G-25 column according to the manufacture instruction.
  2. Load the reaction mixture (From "Run conjugation reaction") to the top of the Sephadex G-25 column.
  3. Add PBS (pH 7.2-7.4) as soon as the sample runs just below the top resin surface.
  4. Add more PBS (pH 7.2-7.4) to the desired sample to complete the column purification. Combine the fractions that contain the desired dye-protein conjugate.
    Note     For immediate use, the dye-protein conjugate need be diluted with staining buffer, and aliquoted for multiple uses.
    Note     For longer term storage, dye-protein conjugate solution need be concentrated or freeze dried. 

Calculators


Common stock solution preparation

Table 1. Volume of DMSO needed to reconstitute specific mass of iFluor® 790 maleimide to given concentration. Note that volume is only for preparing stock solution. Refer to sample experimental protocol for appropriate experimental/physiological buffers.

0.1 mg0.5 mg1 mg5 mg10 mg
1 mM69.451 µL347.256 µL694.512 µL3.473 mL6.945 mL
5 mM13.89 µL69.451 µL138.902 µL694.512 µL1.389 mL
10 mM6.945 µL34.726 µL69.451 µL347.256 µL694.512 µL

Molarity calculator

Enter any two values (mass, volume, concentration) to calculate the third.

Mass (Calculate)Molecular weightVolume (Calculate)Concentration (Calculate)Moles
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Spectrum


Open in Advanced Spectrum Viewer
spectrum

Spectral properties

Correction Factor (260 nm)0.1
Correction Factor (280 nm)0.09
Extinction coefficient (cm -1 M -1)2500001
Excitation (nm)787
Emission (nm)812
Quantum yield0.131

Product family


NameExcitation (nm)Emission (nm)Extinction coefficient (cm -1 M -1)Quantum yieldCorrection Factor (260 nm)Correction Factor (280 nm)
iFluor® 350 maleimide3454502000010.9510.830.23
iFluor® 488 maleimide4915167500010.910.210.11
iFluor® 555 maleimide55757010000010.6410.230.14
iFluor® 647 maleimide65667025000010.2510.030.03
iFluor® 680 maleimide68470122000010.2310.0970.094
iFluor® 700 maleimide69071322000010.2310.090.04
iFluor® 750 maleimide75777927500010.1210.0440.039
iFluor® 800 maleimide80182025000010.1110.030.08
iFluor® 810 maleimide81182225000010.0510.090.15
iFluor® 820 maleimide8228502500001-0.110.16
iFluor® 860 maleimide8538782500001-0.10.14
iFluor® 532 maleimide5375609000010.6810.260.16
iFluor® 594 maleimide58860418000010.5310.050.04
iFluor® 405 maleimide4034273700010.9110.480.77
iFluor® 430 maleimide4334984000010.7810.680.3
iFluor® 568 maleimide56858710000010.5710.340.15
iFluor® 633 maleimide64065425000010.2910.0620.044
iFluor® 450 maleimide4515024000010.8210.450.27
iFluor® 790 Styramide *Superior Replacement for Alexa Fluor 790 tyramide*78781225000010.1310.10.09
iFluor® 460 maleimide468493800001~0.810.980.46
iFluor® 665 maleimide667692110,00010.2210.120.09
iFluor® 546 maleimide54155710000010.6710.250.15
iFluor® 840 maleimide8368792000001-0.20.09
iFluor® 770 maleimide77779725000010.160.090.08
iFluor® 780 maleimide78480825000010.1610.130.12
iFluor® 830 maleimide830867----
iFluor® 514 maleimide5115277500010.8310.2650.116
iFluor® 660 maleimide66367825000010.2610.070.08
iFluor® 670 maleimide67168220000010.5510.030.033
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Citations


View all 1 citations: Citation Explorer
Nanovesicle delivery to the liver via retinol binding protein and platelet-derived growth factor receptors: how targeting ligands affect biodistribution
Authors: Hsu, Ching-Yun and Chen, Chun-Han and Aljuffali, Ibrahim A and Dai, You-Shan and Fang, Jia-You
Journal: Nanomedicine (2017)

References


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A target cell-specific activatable fluorescence probe for in vivo molecular imaging of cancer based on a self-quenched avidin-rhodamine conjugate
Authors: Hama Y, Urano Y, Koyama Y, Kamiya M, Bernardo M, Paik RS, Shin IS, Paik CH, Choyke PL, Kobayashi H.
Journal: Cancer Res (2007): 2791
Fluorescence imaging in vivo: recent advances
Authors: Rao J, Dragulescu-Andrasi A, Yao H.
Journal: Curr Opin Biotechnol (2007): 17
Ex vivo fluorescence imaging of normal and malignant urothelial cells to enhance early diagnosis
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Journal: Photochem Photobiol (2007): 1157
In vivo monitoring the fate of Cy5.5-Tat labeled T lymphocytes by quantitative near-infrared fluorescence imaging during acute brain inflammation in a rat model of experimental autoimmune encephalomyelitis
Authors: Berger C, Gremlich HU, Schmidt P, Cannet C, Kneuer R, Hiest and P, Rausch M, Rudin M.
Journal: J Immunol Methods (2007): 65
A protocol for imaging alternative splicing regulation in vivo using fluorescence reporters in transgenic mice
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Journal: Nat Protoc (2007): 2166
In vivo imaging of the bronchial wall microstructure using fibered confocal fluorescence microscopy
Authors: Thiberville L, Moreno-Swirc S, Vercauteren T, Peltier E, Cave C, Bourg Heckly G.
Journal: Am J Respir Crit Care Med (2007): 22
In Vivo Fluorescence Microscopic Imaging for Dynamic Quantitative Assessment of Intestinal Mucosa Permeability in Mice
Authors: Szabo A, Vollmar B, Boros M, Menger MD.
Journal: J Surg Res. (2007)
In vivo spectral fluorescence imaging of submillimeter peritoneal cancer implants using a lectin-targeted optical agent
Authors: Hama Y, Urano Y, Koyama Y, Kamiya M, Bernardo M, Paik RS, Krishna MC, Choyke PL, Kobayashi H.
Journal: Neoplasia (2006): 607
In vivo imaging of green fluorescent protein-expressing cells in transgenic animals using fibred confocal fluorescence microscopy
Authors: Al-Gubory KH, Houdebine LM.
Journal: Eur J Cell Biol (2006): 837
In vivo near-infrared fluorescence imaging of integrin alphavbeta3 in an orthotopic glioblastoma model
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Journal: Mol Imaging Biol (2006): 315