-
E-mail
1415268707@qq.com
-
Phone
13764885152
-
Address
Room 4597, Building 6, No. 1150 Lanfeng Road, Fengxian District, Shanghai
Shanghai Jintong Instrument Co., Ltd
From Basic to Advanced: Leica Inverted Microscope Operation Guide
Date: 2025-07-18Read: 0
Leica inverted microscopeWith its flexible objective lens configuration and adaptable stage design for multiple samples, it has become a research tool in fields such as cell culture, tissue sectioning, and materials science. Mastering its operational skills can not only improve experimental efficiency, but also extend the lifespan of the instrument. The following provides a step-by-step explanation from basic operations to advanced techniques.
1、 Basic Operations: Standardized Startup and Sample Observation
1. Startup and environment preparation
After connecting the power supply, turn on the power switch of the microscope host. After the eyepiece field of view lights up, turn on the fluorescent light source (if fluorescence observation is required). Attention: Fluorescent light sources need to be preheated for 5-10 minutes to achieve stable light intensity and avoid frequent switching to extend bulb life.
Adjust the microscope base to a stable state to avoid vibration affecting imaging (it is recommended to place it on a shock-absorbing table or thick rubber pad).
2. Sample placement and focusing
Place the culture dish, porous plate, or glass slide in the center of the stage, ensuring that the sample is directly below the objective lens (with the inverted microscope objective facing up and the sample placed from above).
First, use a low magnification objective lens (such as 4X or 10X) for coarse focusing: slowly lower the objective lens to approach the sample using the coarse adjustment knob (be careful not to touch it), and then slowly rotate it in the opposite direction until the image is clear. When switching high magnification objective lenses (such as 20X, 40X), it is necessary to fine tune the knob. Due to the short working distance of the objective lens, the operation needs to be gentler.
3. Optical path and imaging adjustment
Adjust the height and aperture size of the spotlight: When observing with a low-power lens, the position of the spotlight is lower and the aperture is increased; When using a high-power lens, it is necessary to raise the spotlight and reduce the aperture to enhance contrast.
Observe through the eyepiece and adjust the coarse/fine focus knob to the clearest image, avoiding excessive focusing that may damage the objective lens or sample.
2、 Advanced skills: Function expansion and precise imaging
1. Fluorescence observation and channel switching
If fluorescence imaging is required, first confirm that the filter block has been switched to the target fluorescence channel (such as DAPI, FITC, TRITC), and operate it through the filter block dial on the side of the microscope.
Adjust the fluorescence excitation intensity: To avoid quenching the fluorescence due to excessive brightness, the light intensity can be gradually increased through voltage adjustment knob or software control (if connected to a computer) until the signal is clear and the background noise is low.
2. Phase difference and DIC imaging
Phase difference observation: Insert a phase difference ring (to match the objective lens, such as a 10X objective lens corresponding to a 10X phase difference ring), adjust the position of the condenser phase plate, and enhance the contrast of unstained samples (such as live cells).
DIC (Differential Interference Contrast) observation: Install a DIC prism and highlight the fine structures of the sample (such as cell cytoskeleton and cell membrane edges) by adjusting the prism polarization angle and light intensity adjustment knob.
3. Image acquisition and parameter optimization
If connecting a digital camera, set the exposure time and gain value through software (such as Leica Application Suite): the exposure time should be shortened under high magnification (to avoid overexposure), and the gain can be appropriately increased for live cell imaging to enhance brightness.
When shooting with multiple channels, it is necessary to separately capture bright field, fluorescence, or phase difference images, and overlay or compare them in software to ensure consistent parameters (such as the same field of view and exposure time).
3、 Maintenance and Precautions
1. Objective lens cleaning: Use specialized lens paper or a balloon to clean the dust on the surface of the objective lens and avoid direct contact with the lens; If contaminated with liquid, gently wipe with anhydrous ethanol.
2. Stage maintenance: Avoid scratching the surface of the stage with sharp samples, and promptly clean any remaining culture medium or liquid after use.
3. Shutdown process: Turn off the fluorescent light source (cool to room temperature before turning off the power), turn the objective lens to the low magnification position (to avoid long-term pressure on the high magnification objective lens), and finally turn off the power of the host.
Leica inverted microscopeFrom basic focusing to advanced imaging, every step of the operation requires a balance between standardization and detail. Mastering its core techniques not only enables efficient acquisition of high-quality images, but also maximizes the performance of the instrument, providing reliable support for scientific research and experimentation.