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Remote-controlled scanning and automated confocal microscopy through focusing using a modified HRT rostock corneal module.

ABSTRACT: To modify the HRT-II Confocal Microscope with Rostock Corneal Module (HRT-RCM) to allow computerized control of the focal plane position (depth) within the cornea.A threaded housing on the HRT-RCM microscope is normally rotated by hand to change the focal plane position within the cornea. This piece was removed to allow the front housing of the microscope to move freely. A linear actuator (Oriel Encoder Mike) was then attached to the side of the microscope and coupled to a drive shaft that was connected to the front housing. The actuator was connected to an Oriel 18011 Encoder Mike controller, which was interfaced to a PC. Software was developed to allow control and display of the focal plane position using this PC, while image acquisition software was run on the HRT-RCM PC. The instrument was tested on one human volunteer.The modified instrument successfully allowed computer-controlled focusing throughout the entire cornea. Through-focus sequences could be collected online and analyzed and reconstructed three dimensionally off-line using modified confocal microscopy through-focusing software.Although this is only a prototype instrument, it significantly improves the examination procedure by allowing completely "hands-free" operation of the HRT-RCM microscope. The data also demonstrate the feasibility of performing quantitative z-axis scans through the full thickness of the cornea with the HRT-RCM. Given the higher contrast images and improved optical sectioning of the HRT-RCM as compared with other instruments, these capabilities could have widespread application.

SUBMITTER: Petroll WM

PROVIDER: S-EPMC2827927 | biostudies-literature | 2009 Nov

REPOSITORIES: biostudies-literature

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Remote-controlled scanning and automated confocal microscopy through focusing using a modified HRT rostock corneal module.

Petroll W Matthew WM Cavanagh H Dwight HD

Eye & contact lens 20091101 6

<h4>Purpose</h4>To modify the HRT-II Confocal Microscope with Rostock Corneal Module (HRT-RCM) to allow computerized control of the focal plane position (depth) within the cornea.<h4>Methods</h4>A threaded housing on the HRT-RCM microscope is normally rotated by hand to change the focal plane position within the cornea. This piece was removed to allow the front housing of the microscope to move freely. A linear actuator (Oriel Encoder Mike) was then attached to the side of the microscope and cou ...[more]

PMID: 19901584

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Project description:ObjectiveIn vivo Corneal Confocal Microscopy (IVCCM) is a validated, non-invasive test for diabetic sensorimotor polyneuropathy (DSP) detection, but its utility is limited by the image analysis time and expertise required. We aimed to determine the inter- and intra-observer reproducibility of a novel automated analysis program compared to manual analysis.MethodsIn a cross-sectional diagnostic study, 20 non-diabetes controls (mean age 41.4±17.3y, HbA1c 5.5±0.4%) and 26 participants with type 1 diabetes (42.8±16.9y, 8.0±1.9%) underwent two separate IVCCM examinations by one observer and a third by an independent observer. Along with nerve density and branch density, corneal nerve fibre length (CNFL) was obtained by manual analysis (CNFLMANUAL), a protocol in which images were manually selected for automated analysis (CNFLSEMI-AUTOMATED), and one in which selection and analysis were performed electronically (CNFLFULLY-AUTOMATED). Reproducibility of each protocol was determined using intraclass correlation coefficients (ICC) and, as a secondary objective, the method of Bland and Altman was used to explore agreement between protocols.ResultsMean CNFLManual was 16.7±4.0, 13.9±4.2 mm/mm2 for non-diabetes controls and diabetes participants, while CNFLSemi-Automated was 10.2±3.3, 8.6±3.0 mm/mm2 and CNFLFully-Automated was 12.5±2.8, 10.9 ± 2.9 mm/mm2. Inter-observer ICC and 95% confidence intervals (95%CI) were 0.73(0.56, 0.84), 0.75(0.59, 0.85), and 0.78(0.63, 0.87), respectively (p = NS for all comparisons). Intra-observer ICC and 95%CI were 0.72(0.55, 0.83), 0.74(0.57, 0.85), and 0.84(0.73, 0.91), respectively (p<0.05 for CNFLFully-Automated compared to others). The other IVCCM parameters had substantially lower ICC compared to those for CNFL. CNFLSemi-Automated and CNFLFully-Automated underestimated CNFLManual by mean and 95%CI of 35.1(-4.5, 67.5)% and 21.0(-21.6, 46.1)%, respectively.ConclusionsDespite an apparent measurement (underestimation) bias in comparison to the manual strategy of image analysis, fully-automated analysis preserves CNFL reproducibility. Future work must determine the diagnostic thresholds specific to the fully-automated measure of CNFL.

Dataset Information

Remote-controlled scanning and automated confocal microscopy through focusing using a modified HRT rostock corneal module.

Publications

Remote-controlled scanning and automated confocal microscopy through focusing using a modified HRT rostock corneal module.

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