In Vivo Confocal Microscopy: A Standard Operating Procedure for the Detection of Demodex Mites at the Eyelid Margin
July 3rd, 2025
In vivo confocal microscopy is a promising imaging technique with high resolution. We present the following procedure for clinicians to detect Demodex at the eyelid margin in vivo.
We are developing a standardized in vivo confocal microscopy protocol for detecting eyelid of demodex mites to expand clinical adoption. We are adjusting the later of standardized IVCM protocol, which limits detection of demodex mites in vivo in clinical practice. It provides noninvasive, painless, and rapid in vivo detection with easy follow up, eliminating appellation pain and the residue mild issues.
[Narrator] To begin, clean the lens of the device using lens cleaning wipes. Use alcohol-soaked cotton balls to disinfect the headrest and chinrest thoroughly. Press the two black levers together and pull the chinrest forward until it catches to ensure it is in the correct horizontal position. Now, turn the adjustment screw to move the camera away from the patient before the examination. Apply approximately 0.02 milliliters of carbomer-based ophthalmic gel on the front surface of the microscope lens of the device. Next, remove a sterile disposable polymethylmethacrylate cap from its protective cover. Press the sides of the cap firmly onto the microscope lens without touching the anterior surface. Then, apply one drop of gel tear substitute on the front surface of the cap. Create a new patient file in the software. Now, apply a drop of proparacaine hydrochloride at the margin of the eyelid to be examined. Move the scanning camera to the left from the operator's point of view. Then, turn the CCD camera objective lens to adjust the control image perpendicularly to the optical axis of the scanner laser camera. Start a new examination using the software. In the Examination Data tab, select Heidelberg Retina Tomograph Cornea from the Device dropdown list and observe the Cornea Module Settings dialog box. Choose FOV 400 from the Field Lens dropdown list and click OK to confirm. Then, wait for the acquisition window to open and verify the display of the scanning laser camera live image and control image. Next, set the factors in the acquisition window. Select 1x from the CCD Zoom Factor dropdown to set the control image size. Check the automatic brightness and auto boxes to enable automatic brightness control. Ensure the Section Scan Type is selected. To acquire a reference image, turn off the automatic brightness control. Then adjust the focal plane with the adjusting pin until the live image darkens, and subsequently, a bright reflection appears, indicating the anterior surface of the polymethylmethacrylate cap. Click Reset in the Focus Position Micrometer section, and wait for the value to reach 0. Then, check the Automatic Brightness box and switch off Auto Reset. Now, adjust the height of the examination table. Place the patient's chin on the chinrest and forehead against the forehead rest. Adjust the chinrest elevation using the black screw so that the patient's upper eyelids align with the red marks on the headrest column. Instruct the patient to look downward toward the fixed light source and maintain a steady gaze. With a cotton swab, flip the upper eyelid and fully expose the eyelash root and palpebral margin. Turn the adjustment screw to align the laser beam reflection at the upper eyelid level. Then, move the scanning laser camera until the upper eyelid margin gently contacts the center of the polymethylmethacrylate cap. Carefully scan the eyelash root from the temporal to nasal side using minute horizontal movements of the palpebral margin. Turn the microscope lens clockwise or counterclockwise to modify the focus and observe entire follicles and roots. Record all suspected demodex images using the foot pedal. After quitting the examination, select Sort by Acquisition Time fast in the Options dialog to display images from temporal to nasal side. In 2024, the number of subjects undergoing in vivo confocal microscopy for demodex detection increased significantly across all age groups compared to 2023, with the highest number observed in the 21 to 30-year age group. The standard operating procedure introduced in 2024 enabled the inclusion of subjects from as young as four years old to over 90 years of age.
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Overview
This article presents a standardized in vivo confocal microscopy protocol for detecting Demodex mites at the eyelid margin. The method is noninvasive and aims to enhance clinical adoption.
Key Study Components
Area of Science
- Ophthalmology
- Imaging Techniques
- Clinical Protocols
Background
- Demodex mites are common parasites affecting eyelid health.
- Traditional detection methods can be invasive and uncomfortable.
- In vivo confocal microscopy offers a high-resolution imaging alternative.
- The study aims to improve detection rates and patient comfort.
Purpose of Study
- To develop a standardized protocol for in vivo detection of Demodex mites.
- To facilitate easier clinical adoption of the imaging technique.
- To provide a noninvasive and rapid detection method.
Methods Used
- Cleaning and disinfecting the microscopy equipment.
- Applying a carbomer-based gel for optimal imaging.
- Utilizing a scanning laser camera to capture images of the eyelid margin.
- Recording and sorting images for analysis.
Main Results
- Increased detection of Demodex mites across various age groups.
- Standardized protocol enabled examination of subjects aged 4 to over 90.
- Significant rise in subjects examined in 2024 compared to 2023.
- Improved patient comfort and ease of follow-up.
Conclusions
- The standardized protocol enhances the detection of Demodex mites.
- In vivo confocal microscopy is a viable option for clinical practice.
- Future studies may expand on the protocol's applications.
