Introduction

The Trinocular Mercury Lamp Fluorescence Microscope utilizes the principle of fluorescence to illuminate the object by using a mercury lamp as a light source. The mercury lamp gives accurate, fast, and reliable results due to its ability to produce bright spectral lines within the visible region of the UV spectrum. 

It has two eyepieces similar to traditional binocular microscopes and contains an additional eyetube for connecting a microscope camera. Which not only captures the real-time images but also records videos. The recorded observations can be saved for future use, allowing detailed and prolonged analysis.

The Trinocular Fluorescence Microscope is highly precise and produces clear, fine images with the slightest division of 2 microns. It also includes a 6V 20W halogen lamp for transmission illumination.

Apparatus

The Trinocular Mercury Lamp Fluorescence Microscope is equipped with two eyepieces (binocular) and an additional third eyepiece tube to connect a microscope camera, making it trinocular. It has a focus system consisting of fine and coarse adjustment knobs. It also contains a quadruple-reversed nosepiece, double-layer mechanical working stage, abbe condenser, collector for placing a halogen lamp, and blue and frosted filters. The light source is provided by a mercury lamp and a halogen lamp. The fluorescence system is composed of a filter system for B Exciton Wavelength: 450~490nm and a filter system for G Exciton Wavelength: 495~555nm.

Principle

It works on the principle of fluorescence that illuminates the sample through excitation and emission wavelengths from the microscope’s light sources. The microscope’s camera captures the images of the specimen or records videos. 

Protocol

• Place the microscope on a solid table
• Place the slide on the working stage and secure it with a clamp
• Turn on the power supply 
• Move the working stage until the slide is at the center of the field of view
• Move the stage up using the coarse adjustment knob 
• View the specimen through the eyepiece using the desired objective lens
• Adjust the fineness and coarseness of the image
• Install the camera in the trinocular head carefully. To install a microscope camera correctly, an adapter (ideally a c-mount adapter) is required that must be affixed to the camera port 
• Capture the images of the specimen or record videos

Applications

The Trinocular Mercury Lamp Fluorescence Microscope is used in various fields to identify and observe specimens. For example, it is used to identify cells, infectious agents, toxins, bioaerosol, suspicious munitions, tumors, microbial aerosols, microscopic organisms, and water quality. Its diverse usage credit goes to the microscope’s mercury lamp which has a wide and visible spectral band range.

Infectious Agents’ Diagnosis

The Trinocular Mercury Lamp Fluorescence Microscope can effectively diagnose various conditions. It can identify soluble toxins and viruses with fluorescent antibodies that are impossible to detect with other microscopes. Sanborn, Heuck, & Aouad (n.d.) used a fluorescence microscope to demonstrate staphylococcal enterotoxin infection in an animal’s body and localized its activity in the brain and other organs where it produced toxic symptoms.

Environment Monitoring

It is often used in public health labs to test oil, water, and air as it has the sensitivity to detect even dead or impaired microorganisms.

Biological Research

The precise location, development, and function of tissues and cells can be detected using fluorochromes and fluorescent antibodies. Subcellular components such as cell wall structure, composition, and nucleic acids may also be marked by fluorochromes, depending on characteristics such as the specimen’s chemical composition, viscosity, and pH.

Strengths 

The Trinocular Mercury Lamp Fluorescence Microscope comes with a high-resolution camera that provides real-time specimen observation, saving time, as there are no risks of wrong observations. Bright and digitally controlled LED light sources and straightforward software enable the production of publication-quality photographs in just a few clicks. It has powerful imaging features to handle time-lapse movies, multiwell plate scanning, picture stitching and tiling, and cell counting. The microscope minimizes the complexity of high-end microscopy without losing performance. 

Summary

• The Trinocular Mercury Lamp Fluorescence Microscope is an adaptation of binocular microscopes since it contains one extra port for placing a microscope camera. Which not only captures real-time images but also records videos.
• It contains 2 eyepieces, an eyepiece tube, objectives, nosepiece, working stage, abbe condenser, collector, filters, mercury lamp, and a halogen lamp.
• It works on the principle of fluorescence that illuminates the sample through excitation and emission wavelengths from the microscope’s light sources.
• Bright and digitally controlled LED light sources and straightforward software enable you to produce amazing, publication-quality photographs in just a few clicks.

References

1. Sanborn, W., Heuck, C., & Aouad, R. (n.d.). Fluorescence microscopy for disease diagnosis and environmental monitoring. Retrieved from https://applications.emro.who.int/dsaf/dsa281.pdf