Novel Multiwavelength Microscopic Scanner for Mouse Imaging

Alencar, Herlen; Mahmood, Umar; Kawano, Yoshihiro; Hirata, Tadashi; Weissleder, Ralph

doi:10.1593/neo.05376

Cited by 60 publications

(47 citation statements)

References 20 publications

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“…The pancreas and surrounding organs were then surveyed using the fiberoptic catheter, in a similar manner to laparoscopic clinical procedures. Finally, the scanned area was also imaged using a confocal laser microscope (IV100) (37), operating at two different optical channels (488-nm excitation for MIP-GFP and 748-nm excitation for E4 ×12 -VT750). This was used to confirm that the observed fluorescent areas did indeed colocalize with MIP-GFP-expressing cells.…”

Section: Methodsmentioning

confidence: 99%

Accurate measurement of pancreatic islet β-cell mass using a second-generation fluorescent exendin-4 analog

Reiner

Thurber

Gaglia³

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

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111

126

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The hallmark of type 1 diabetes is autoimmune destruction of the insulin-producing β-cells of the pancreatic islets. Autoimmune diabetes has been difficult to study or treat because it is not usually diagnosed until substantial β-cell loss has already occurred. Imaging agents that permit noninvasive visualization of changes in β-cell mass remain a high-priority goal. We report on the development and testing of a near-infrared fluorescent β-cell imaging agent. Based on the amino acid sequence of exendin-4, we created a neopeptide via introduction of an unnatural amino acid at the K 12 position, which could subsequently be conjugated to fluorophores via bioorthogonal copper-catalyzed click-chemistry. Cell assays confirmed that the resulting fluorescent probe (E4 ×12 -VT750) had a high binding affinity (∼3 nM). Its in vivo properties were evaluated using high-resolution intravital imaging, histology, whole-pancreas visualization, and endoscopic imaging. According to intravital microscopy, the probe rapidly bound to β-cells and, as demonstrated by confocal microscopy, it was internalized. Histology of the whole pancreas showed a close correspondence between fluorescence and insulin staining, and there was an excellent correlation between imaging signals and β-cell mass in mice treated with streptozotocin, a β-cell toxin. Individual islets could also be visualized by endoscopic imaging. In short, E4 ×12 -VT750 showed strong and selective binding to glucose-like peptide-1 receptors and permitted accurate measurement of β-cell mass in both diabetic and nondiabetic mice. This near-infrared imaging probe, as well as future radioisotope-labeled versions of it, should prove to be important tools for monitoring diabetes, progression, and treatment in both experimental and clinical contexts.

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Section: Methodsmentioning

confidence: 99%

Accurate measurement of pancreatic islet β-cell mass using a second-generation fluorescent exendin-4 analog

Reiner

Thurber

Gaglia³

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

111

126

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show abstract

“…A prototype multichannel upright laser-scanning fluorescent microscope (IV100; Olympus, Tokyo, Japan) with a custom-designed stage and scanning unit for intravital observations was used (Alencar et al, 2005). The stage was equipped with a heating plate regulated by a thermostat (37°C).…”

Section: Methodsmentioning

confidence: 99%

Bimodal Viral Vectors andIn VivoImaging Reveal the Fate of Human Neural Stem Cells in Experimental Glioma Model

Shah¹,

Hingtgen²,

Kasmieh³

et al. 2008

J. Neurosci.

125

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“…Another much worthy improvement of the technique involves objectives [9,10,77], or graded-index (GRIN) lenses [78,79], with extremely slim diameter tips (about 1 mm) that require a much tinier thoracotomy. The reduced optical efficiency of these lenses in terms of field of view and numerical aperture is counterbalanced by a much reduced invasiveness.…”

Section: Imagingmentioning

confidence: 99%

Intravital microscopy of the lung: minimizing invasiveness

Fiole

Tournier

2016

Journal of Biophotonics

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In vivo microscopy has recently become a gold standard in lung immunology studies involving small animals, largely benefiting from the democratization of multiphoton microscopy allowing for deep tissue imaging. This technology represents currently our only way of exploring the lungs and inferring what happens in human respiratory medicine. The interest of lung in vivo microscopy essentially relies upon its relevance as a study model, fulfilling physiological requirements in comparison with in vitro and ex vivo experiments. However, strategies developed in order to overcome movements of the thorax caused by breathing and heartbeats remain the chief drawback of the technique and a major source of invasiveness. In this context, minimizing invasiveness is an unavoidable prerequisite for any improvement of lung in vivo microscopy. This review puts into perspective the main techniques enabling lung in vivo microscopy, providing pros and cons regarding invasiveness.

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Novel Multiwavelength Microscopic Scanner for Mouse Imaging

Cited by 60 publications

References 20 publications

Accurate measurement of pancreatic islet β-cell mass using a second-generation fluorescent exendin-4 analog

Accurate measurement of pancreatic islet β-cell mass using a second-generation fluorescent exendin-4 analog

Bimodal Viral Vectors andIn VivoImaging Reveal the Fate of Human Neural Stem Cells in Experimental Glioma Model

Intravital microscopy of the lung: minimizing invasiveness

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