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WIREs Nanomed Nanobiotechnol
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Aptamers: turning the spotlight on cells

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Abstract This article is a review of the development and application of aptamer probes for cell imaging. Aptamers selected against whole cells have been modified with different fluorescent dyes and nanomaterials, such as gold nanoparticles, quantum dots, and superparamagnetic iron oxide, for their use as imaging probes of live cells. These probes have been successfully used for cell imaging both in vitro and in vivo by optical imaging, magnetic resonance imaging (MRI), computed tomography (CT), and positron‐emission tomography (PET). In this article, we discuss the development of different aptamer‐based probes currently available for imaging of live cells and their applications in the biomedical field. WIREs Nanomed Nanobiotechnol 2011 3 328–340 DOI: 10.1002/wnan.133 This article is categorized under: Diagnostic Tools > In Vitro Nanoparticle-Based Sensing Diagnostic Tools > In Vivo Nanodiagnostics and Imaging Nanotechnology Approaches to Biology > Nanoscale Systems in Biology

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Confocal microscopy imaging of cells treated with quantum dots (QD)–aptamer conjugates. (a) A single incubation of QD605‐TTA1, QD655‐AS1411, or QD705‐MUC‐1 was applied to PC‐3, HeLa, CHO, C6, and NPA cells, and confocal images were obtained. Each image was compared with the corresponding QD–control aptamers (column 1: QD‐TTA1, column 2: QD‐TTA1 control, column 3: QD‐AS1411, column 4: QD‐AS1411 control, column 5: QD‐MUC‐1, column 6: QD‐MUC‐1 control). (b) Multiplex imaging of cancer cells treated simultaneously with three different types of QD‐conjugated aptamers. Single images for QD‐TTA1 (605 nm, light green, column 1), QD‐AS1411 (655 nm, red, column 2), and QD‐MUC‐1 (705 nm, violet, column 3), dual images for QD‐AS1411 and QD‐TTA1 (column 4, yellow for colocalization), QD‐TTA1 and QD‐MUC‐1 (column 5, light green for colocalization), and QD‐AS1411 and QD‐MUC‐1 (column 6, violet for colocalization), and a triple image for QD‐AS1411, QD‐TTA1, and QD‐MUC‐1 (column 7, white for colocalization) were acquired from PC‐3, HeLa, CHO, C6, and NPA cells. All figures are merged with the 40, 6‐diamidino‐2‐phenylindole (DAPI) image (nucleus staining, 460 nm) and cellular morphology. (Reprinted with permission from Ref 36. Copyright 2009 Wiley‐VCH Verlag GmbH & Co [Small])

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Fluorescence confocal images and optical images of MEAR cells (a) and BNL cells (b) stained by unselected library (top), aptamers TLS9a (middle), and TLS11a (bottom) labeled with Fluorescein isothiocyanate (FITC). In each picture, left is the fluorescence images and right is the optical images for MEAR cells and BNL cells, respectively. (Reprinted with permission from Ref 7. Copyright 2008 American Chemical Society publications [Analytical Chemistry])

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Comparative efficacy study in Ramos s.c. xenograft nude mice. (a) Optical and fluorescence imaging of tumors in live mice using Cy5 dye and Cy5‐labeled aptamers. Ramos cells were injected subcutaneously on the backs of BALB/c nude mice 2–3 weeks before imaging. Cy5 dye, Cy5‐labeled Sgc8a, or Cy5‐labeled TD05 was subsequently injected intravenously through the tail vein, and fluorescence images of the dorsal side of live mice were taken at various time points after injection. Images of a, b, and c represent the mouse photos, the fluorescence images before and 3.5 h after injection of probes, respectively. The fluorescence images had been processed by the spectrum separation method and Image J software (time of exposure for every image is 4 seconds). (b) Quantification of the overall signal‐to‐background ratio for Cy5‐labeled TD05 and Cy5‐labeled Sgc8a. Signal‐to‐background ratios were calculated from images in Figure 4a based on the fluorescence signals of tumor relative to background measured for a region on the back (red circles) by using Image J software. The number of mice used to derive statistical information for each probe is five. Data represent mean ± standard error. (Reprinted with permission from Ref 60. Copyright 2010 Wiley‐VCH Verlag GmbH & Co [Chemistry: An Asian Journal])

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The CD30 aptamer‐mediated immunostaining in different tissues. (a) Immunostaining of necrotic tissue. Hematoxylin and eosin (H and E) staining of a CD30‐negative peripheral T‐cell lymphoma revealed the presence of necrosis in the upper left portion and viable lymphoma cells in the lower right portion of the tissue section. Probing of the tissue with the standard CD30 antibody showed nonspecific staining on cell debris/‘ghost’ cells (indicated by arrows) within the necrotic area, although no reaction with adjacent viable lymphoma cells that do not express CD30 was noted. In contrast, immunostaining with the synthetic CD30 aptamer probe showed no nonspecific background staining in the necrotic areas or in the adjacent viable lymphoma cells. (b) Comparison of the immunostaining by the synthetic CD30 aptamer probe and the CD30 antibody on sections of different CD30‐negative tissues. Formalin‐fixed and paraffin‐embedded tissue sections of normal lymph node, lung, stomach, kidney, adrenal gland, and a metastatic carcinoma of the lung were prepared with H and E stain (top row), immunostained with the CD30 aptamer probe (middle row), or immunostained with the CD30 antibody as a standard control (bottom row). (Reprinted with permission from Ref 50. Copyright 2010 Macmillan Publishers Ltd. [Modern Pathology])

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Confocal microscopy images of individual NP–aptamer conjugates with the three different cells (Toledo, CEM, and Ramos): (a) NP (FAM)‐T1, (b) NP (FAM‐R6G)‐sgc8, and (c) NP (FAM‐R6G‐ROX)‐TDO5. (Reprinted with permission from Ref 36. Copyright 2009 American Chemical Society publications [Analytical Chemistry])

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