Fluorescence diffuse tomography of small animals with DsRed2 fluorescent protein

Turchin, Ilya V.; Plehanov, Vladimir I.; Orlova, Anna; Kamenskiy, V. A.; Kleshnin, Mikhail; Shirmanova, Marina V.; Shakhova, Natalia M.; Balalaeva, Irina V.; Savitskiy, A. P.

doi:10.1134/s1054660x06050021

Cited by 12 publications

(15 citation statements)

References 17 publications

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“…Trans-illumination data have been used in planar or three-dimensional tomographic imaging for imaging deep-seated fluorescence bio-distribution with increased sensitivity and resolution compared to epi-illumination imaging in phantoms and in vivo 12–14. Imaging of fluorescence proteins have been similarly showcased 13,15. However, conventional fluorescent proteins emit in the visible, where light absorption by tissue is strong.…”

Section: Introductionmentioning

confidence: 99%

Performance of the red-shifted fluorescent proteins in deep-tissue molecular imaging applications

et al. 2008

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The discovery of new fluorescent proteins (FPs) that emit in the far-red part of the spectrum, where light absorption from tissue is significantly lower than in the visible, offers the possibility for noninvasive biological interrogation at the entire organ or small animal level in vivo. The performance of FPs in deep-tissue imaging depends not only on their optical characteristics, but also on the wavelength-dependent tissue absorption and the depth of the fluorescence activity. In order to determine the optimal choice of FP and illumination wavelength we compared the performance of five of the most promising FPs, i.e. tdTomato, mCherry, mRaspberry, mPlum, and Katushka. We experimentally measured the signal strength through mice and employed theoretical predictions to obtain an understanding on the performance of different illumination scenarios, especially as it pertains to tomographic imaging. It was found that the appropriate combination of red-shifted proteins and illumination wavelengths can improve detection sensitivity in small animals by at least 2 orders of magnitude compared to Green FP (GFP). It is also shown that the steep attenuation change of the hemoglobin spectrum around the 600nm range may significantly affect the detection sensitivity and necessitates the careful selection of illumination wavelengths for optimal imaging performance.

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Section: Introductionmentioning

confidence: 99%

Performance of the red-shifted fluorescent proteins in deep-tissue molecular imaging applications

et al. 2008

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“…Fluorescence tomography-time domain ͑TD͒, frequency domain ͑FD͒, and continuous wave ͑CW͒-use reconstruction algorithms that account for the effects of diffuse light propagation in tissue. [8][9][10][11][12][13][14][15][16][17][18][19][20][21] Appropriate theoretical model of photon propagation in tissues with the corresponding mathematical inversion permits one to determine, with high resolution, the real boundaries of tumors located deep in animals. These techniques are used mainly to investigate the distribution of near-infrared fluorescent probes.…”

Section: Introductionmentioning

confidence: 99%

“…Only a few works have been devoted to fluorescence diffuse tomography ͑FDT͒ with excitation in visible light. [17][18][19][20][21] Highly sensitive systems are required to detect fluorescent light at large depths ͑Ͼ5 to 7 mm͒ due to the high absorption rate of biological tissue in this spectral range. Photomultiplier tubes ͑PMTs͒ and cooled charge-coupled devices ͑CCD͒ are traditionally used for fluorescence detection.…”

Section: Introductionmentioning

confidence: 99%

Fluorescence diffuse tomography for detection of red fluorescent protein expressed tumors in small animals

Turchin¹,

Kamensky²,

Plehanov³

et al. 2008

J. Biomed. Opt.

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A fluorescence diffuse tomography (FDT) setup for monitoring tumor growth in small animals has been created. In this setup an animal is scanned in the transilluminative configuration by a single source and detector pair. To remove stray light in the detection system, we used a combination of interferometric and absorption filters. To reduce the scanning time, an experimental animal was scanned using the following algorithm: (1) large-step scanning to obtain a general view of the animal (source and detector move synchronously); (2) selection of the fluorescing region; and (3) small-step scanning of the selected region and different relative shifts between the source and detector to obtain sufficient information for 3D reconstruction. We created a reconstruction algorithm based on the Holder norm to estimate the fluorophore distribution. This algorithm converges to the solution with a minimum number of fluorescing zones. The use of tumor cell lines transfected with fluorescent proteins allowed us to conduct intravital monitoring studies. Cell lines of human melanomas Mel-P, Mel-Ibr, Mel-Kor, and human embryonic kidney HEK293 Phoenix were transfected with DsRed-Express and Turbo-RFP genes. The emission of red fluorescent proteins (RFPs) in the long-wave optical range permits detection of deep-seated tumors. In vivo experiments were conducted immediately after subcutaneous injection of fluorescing cells into small animals.

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“…Different types of fluorescent tomography, time-domain (TD), frequency-domain (FD), and continuous wave (CW) accounts for the diffusive propagation of photons in tissue [4][5][6][7][8][9][10][11][12]. Using theoretical models of photon propagation in tissues and subsequent mathematical inversion permit one to determine with high resolution real boundaries of tumors located deep in animal.…”

Section: Introductionmentioning

confidence: 99%

“…These techniques were used for investigation of the distribution of near-infrared fluorescent probes. Only a few works are devoted to fluorescence tomography in visible light [11][12]. Highly sensitive systems are required for detection of fluorescence light from large depths (>3-4 mm) due to high absorption of biological tissue in this spectrum range.…”

Section: Introductionmentioning

confidence: 99%

Fluorescence diffuse tomography for detection of RFP-expressed tumors in small animals

Turchin¹,

Savitsky

Kamensky³

et al. 2007

SPIE Proceedings

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Capabilities of tumor detection by different optical methods can be significantly improved by labeling of tumors with fluorescent markers. Creation of tumor cell lines transfected with fluorescent proteins provides the possibility not only to detect tumor, but also to conduct the intravital monitoring studies. Cell lines of human melanomas Mel-P, Mel-Kor and human embryonic kidney HEK-293 Phoenix were transfected with DsRed-Express and Turbo-RFP genes. Emission of RFP in the long-wave optical range permits detection of the deeply located tumors, which is essential for whole-body imaging. Only special tools for turbid media imaging, such as fluorescent diffusion tomography (FDT), enable noninvasive investigation of the internal structure of biological tissue. FDT setup for monitoring of tumor growth in small animals has been created. An animal is scanned in the transilluminative configuration by low-frequency modulated light (1 kHz) from Nd:YAG laser with second harmonic generation at the 532 nm wavelength. An optimizing algorithm for scanning of an experimantal animal is suggested. In vivo experiments were conducted immediately after the subcutaneously injection of fluorescing cells into small animals. It was shown that FDT method allows to detect the presence of fluorescent cells in small animals and can be used for monitoring of tumor growth and anticancer drug responce. Downloaded From: http://proceedings.spiedigitallibrary.org/ on 06/25/2016 Terms of Use: http://spiedigitallibrary.org/ss/TermsOfUse.aspx SPIE-OSA/ Vol. 6626 66260R-5 Downloaded From: http://proceedings.spiedigitallibrary.org/ on 06/25/2016 Terms of Use: http://spiedigitallibrary.org/ss/TermsOfUse.aspxFigure 6. Fluorescent images, obtained in transillumination configuration of source and detector of a nude mice after injection of НЕК293-TurboRFР cells. The images obtaind (from left to right): immediately after injection, 24 hours later, and 48 hours later. Number of cells 1.7 million, suspension volume 50 µl. SPIE-OSA/ Vol. 6626 66260R-6 Downloaded From: http://proceedings.spiedigitallibrary.org/ on 06/25/2016 Terms of Use: http://spiedigitallibrary.org/ss/TermsOfUse.aspx

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Fluorescence diffuse tomography of small animals with DsRed2 fluorescent protein

Cited by 12 publications

References 17 publications

Performance of the red-shifted fluorescent proteins in deep-tissue molecular imaging applications

Performance of the red-shifted fluorescent proteins in deep-tissue molecular imaging applications

Fluorescence diffuse tomography for detection of red fluorescent protein expressed tumors in small animals

Fluorescence diffuse tomography for detection of RFP-expressed tumors in small animals

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