Low cost, high performance, self-aligning miniature optical systems

Kester, Robert T.; Christenson, Todd; Kortum, Rebecca Richards; Tkaczyk, Tomasz S.

doi:10.1364/ao.48.003375

Cited by 22 publications

(17 citation statements)

References 13 publications

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“…GRIN lenses offer a straightforward and economical way to increase spatial resolution, though their susceptibility to optical aberrations and limited NA is well recognized. If GRIN lens performance is inadequate for a particular application, hybrid GRIN / spherical lens objectives 9 or miniature objective lens assemblies [10][11] can be employed.…”

Section: Discussionmentioning

confidence: 99%

High-resolution Fiber-optic Microendoscopy for <em>in situ</em> Cellular Imaging

Pierce¹,

Yu²,

Richards‐Kortum³

2011

JoVE

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Many biological and clinical studies require the longitudinal study and analysis of morphology and function with cellular level resolution. Traditionally, multiple experiments are run in parallel, with individual samples removed from the study at sequential time points for evaluation by light microscopy. Several intravital techniques have been developed, with confocal, multiphoton, and second harmonic microscopy all demonstrating their ability to be used for imaging in situ 1 . With these systems, however, the required infrastructure is complex and expensive, involving scanning laser systems and complex light sources. Here we present a protocol for the design and assembly of a high-resolution microendoscope which can be built in a day using off-the-shelf components for under US$5,000. The platform offers flexibility in terms of image resolution, field-of-view, and operating wavelength, and we describe how these parameters can be easily modified to meet the specific needs of the end user.We and others have explored the use of the high-resolution microendoscope (HRME) in in vitro cell culture [2][3][4][5] , in excised 6 and living animal tissues 2,5 , and in human tissues in vivo 2,7 . Users have reported the use of several different fluorescent contrast agents, including proflavine [2][3][4] , benzoporphyrin-derivative monoacid ring A (BPD-MA) 5 , and fluoroscein 6,7 , all of which have received full, or investigational approval from the FDA for use in human subjects. High-resolution microendoscopy, in the form described here, may appeal to a wide range of researchers working in the basic and clinical sciences. The technique offers an effective and economical approach which complements traditional benchtop microscopy, by enabling the user to perform high-resolution, longitudinal imaging in situ. Video LinkThe video component of this article can be found at https://www.jove.com/video/2306/ Protocol Microendoscope AssemblyThe high-resolution microendoscope described here (figure 1a) should be considered as a base configuration with several variations possible in assembly and application. We describe in detail here an embodiment of the platform which is designed to be used with proflavine as a fluorescent contrast agent. Proflavine is a bright nuclear stain with peak absorption and emission wavelengths of 445 nm and 515 nm respectively. The use of other contrast agents will require the user to select excitation, emission, and dichroic filters appropriately. Several elements of the high-resolution microendoscope are generic and may be obtained from multiple vendors. For example, optomechanical positioning components are available from Thorlabs, Newport, Linos among others. Compact CCD cameras are available from companies including Point Grey Research, Prosilica, and Retiga; camera sensitivity should be chosen with consideration of the brightness of the fluorophore to be used, as well as the desired frame rate. High-power light emitting diodes (LEDs) may be obtained from Luxeon, Cree, and Nichia among others. ...

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Section: Discussionmentioning

confidence: 99%

High-resolution Fiber-optic Microendoscopy for <em>in situ</em> Cellular Imaging

Pierce¹,

Yu²,

Richards‐Kortum³

2011

JoVE

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“…1a), each of which used different materials and fabrication techniques 9,18,19,22 . These lens systems have similar specifications (numerical aperture of 1.0, field-of-view of 250 m, and outer diameter less than 10 mm), yet they differ significantly in their cost and potential for mass production (Table 1).…”

Section: Discussionmentioning

confidence: 99%

“…The hybrid objective lens systems were fabricated and assembled as described previously 19 . As a quantitative measure of optical quality, the Strehl ratio was determined by using the slanted edge modulation transfer function technique.…”

Section: Methodsmentioning

confidence: 99%

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Optical Systems for Point-of-care Diagnostic Instrumentation: Analysis of Imaging Performance and Cost

et al. 2013

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One of the key elements in point-of-care (POC) diagnostic test instrumentation is the optical system required for signal detection and / or imaging. Many tests which use fluorescence, absorbance, or colorimetric optical signals are under development for management of infectious diseases in resource limited settings, where the overall size and cost of the device is of critical importance. At present, high-performance lenses are expensive to fabricate and difficult to obtain commercially, presenting barriers for developers of in vitro POC tests or microscopic image-based diagnostics. We recently described a compact “hybrid” objective lens incorporating both glass and plastic optical elements, with a numerical aperture of 1.0 and field-of-view of 250 m. This design concept may potentially enable mass-production of high-performance, low-cost optical systems which can be easily incorporated in the readout path of existing and emerging POC diagnostic assays. In this paper, we evaluate the biological imaging performance of these lens systems in three broad POC diagnostic application areas; (1) bright field microscopy of histopathology slides, (2) cytologic examination of blood smears, and (3) immunofluorescence imaging. We also break down the fabrication costs and draw comparisons with other miniature optical systems. The hybrid lenses provided images with quality comparable to conventional microscopy, enabling examination of neoplastic pathology and infectious parasites including malaria and cryptosporidium. We describe how these components can be produced at below $10 per unit in full-scale production quantities, making these systems well suited for use within POC diagnostic instrumentation.

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“…4(b) allow the lenses to be assembled with no alignment necessary (a zero alignment technique 14,17,19 ). The alignment features uniquely define the position of the lenses laterally and axially, allowing them to be stacked one atop the other and glued in place.…”

Section: Fabrication Of the Endomicroscopementioning

confidence: 99%

Needle-based fluorescence endomicroscopy via structured illumination with a plastic, achromatic objective

et al. 2013

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Abstract. In order to diagnose cancer, a sample must be removed, prepared, and examined under a microscope, which is expensive, invasive, and time consuming. Fiber optic fluorescence endomicroscopy, where an image guide is used to obtain high-resolution images of tissue in vivo, has shown promise as an alternative to conventional biopsies. However, the resolution of standard endomicroscopy is limited by the fiber bundle sampling frequency and out-of-focus light. A system is presented which incorporates a plastic, achromatic objective to increase the sampling and which provides optical sectioning via structured illumination to reject background light. An image is relayed from the sample by a fiber bundle with the custom 2.1-mm outer diameter objective lens integrated to the distal tip. The objective is corrected for the excitation and the emission wavelengths of proflavine (452 and 515 nm). It magnifies the object onto the fiber bundle to improve the system's lateral resolution by increasing the sampling. The plastic lenses were fabricated via single-point diamond turning and assembled using a zero alignment technique. Ex vivo images of normal and neoplastic murine mammary tissues stained with proflavine are captured. The system achieves higher contrast and resolves smaller features than standard fluorescence endomicroscopy.

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Low cost, high performance, self-aligning miniature optical systems

Cited by 22 publications

References 13 publications

High-resolution Fiber-optic Microendoscopy for <em>in situ</em> Cellular Imaging

High-resolution Fiber-optic Microendoscopy for <em>in situ</em> Cellular Imaging

Optical Systems for Point-of-care Diagnostic Instrumentation: Analysis of Imaging Performance and Cost

Needle-based fluorescence endomicroscopy via structured illumination with a plastic, achromatic objective

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