Fluorescence Microscopy and Its Applications in the Chemical Industry

Fluorescence Microscopy and Applications in the Chemical Industry

Since 1989, An-ka Analiz Kalite Kontrol Cihazları San. Tic. Ltd. Şti. has grown steadily under the vision and leadership of founder İbrahim Bağış, establishing itself as one of the leading firms in the sector through product and service quality that lives up to the promise of "quality" in its name across all areas in which it operates. Our product groups include laboratory-type glassware washing, drying and disinfection equipment, vacuum pumps and measurement systems, helium gas leak detectors, microscopes, digital microscopes and microscope image transfer systems, atmosphere-controlled and vacuum high-temperature furnaces, and laboratory quality control test equipment. We serve in these product groups through long-standing partnerships with the sector's leading brands. Microscopes are generally optical instruments used for the examination of living or non-living entities not visible to the naked eye. This optical instrument has different types for different methods and purposes. The type of microscope we will discuss here is known as fluorescence microscopy.

First, let us explain the meaning of some technical terms:

Fluorescence: The phenomenon whereby objects reflect the light rays striking them back at wavelengths different from those of the incident light is called fluorescence. Excitation: If the excitation energy comes from a light source, this is called excitation light. Emission: The light emitted upon return is called emission. Fluorophore: Molecules that absorb excitation light at a specific wavelength (typically UV) and emit light at a longer wavelength after a brief delay.

Why is Fluorescence Microscopy Needed?

The fluorescence of an object is obtained either through autofluorescence or secondary (secondary fluorescence) obtained as a result of staining the object with certain dyes. In this way, it becomes possible to examine details of structures that cannot be seen with normal optical microscopy. In fluorescence microscopes, fluorophore materials excited by light of the appropriate wavelength emit light at a longer wavelength. This light emission decays in measurable times of milliseconds. This type of imaging technique is called fluorescence imaging. Microscopes using these techniques are called fluorescence microscopes. The principle in fluorescence microscopes is outlined roughly as follows in the image below. Light from a light source passes through an excitation filter, exciting the sample and causing light to be emitted at a specific wavelength. The most important development regarding fluorescence microscopy is the awarding of the Nobel Prize in Chemistry to American scientists Eric Betzig and William E. Moerner and German scientist Stefan W. Hell. The scientists sharing the prize—Eric Betzig (Howard Hughes Health Institute, USA), William E. Moerner (Stanford University, USA), and Stefan W. Hell—were recognized for their work on developing "high-resolution fluorescence microscopy".

Applications of Fluorescence Microscopy

Several examples can be given of the industrial use of fluorescence microscopy: In the dyeing industry, dyes or detergents used produce various effects on samples. It is more reliable to examine the reasons for these effects using fluorescence microscopy. When a fabric washed with an appropriate fluorescent dye is examined under a fluorescence microscope: • If there is a mechanical defect, fibers appear pink, • If damaged by alkali, yellow-pink, • If damaged by acid, blue-pink color will appear. A fiber with no defects will be colorless. In studies conducted with a Soptop brand RX50 model microscope, the opportunity to work with different dyes was made possible thanks to the microscope's superior multiple filter holder feature. Images captured with fluorescence microscopes can be transferred to a computer through special image transfer systems. The image from the microscope was transferred to the computer with the Soptop brand OD 140B CCD model image transfer system. Cameras used in fluorescence microscopes must specifically have CCD characteristics. In low-emission samples, CCD-equipped cameras demonstrate greater resolution and color transfer success. In another study conducted with the Soptop brand RX50 model microscope, morphological examinations were performed on a mixture of bitumen and γ-PPR polymer. Detailed information about the penetration of the mixture was obtained thanks to the UV filter. In yet another study performed with the Soptop brand RX 50 model microscope, silica particles were titanium-coated and subsequently stained with Rhodamine dye. This study examined the imaging differences between inverted microscopes and fluorescence microscopes. This examination was conducted to visualize how nanoparticles are distributed within the system, and successful results were obtained. Coating manufacturers use fluorescence microscopes to examine coating quality and distribution. Similarly, in the healthcare sector, fluorescence microscopy is used for bacterial identification, imaging cancer cells, and in every field where imaging through staining techniques is needed.

Important Considerations When Selecting a Fluorescence Microscope

There are three important considerations regarding fluorescence microscopy. The first and most important is objective quality, the second is the quality of the selected fluorescence filter, and the third is the fluorescent dye used for staining. If any one of these three options is poor, it results in dim images, very brief light emission, or no emission at all. For this reason, in the field of fluorescence microscopy, we provide service with Soptop brand premium model microscopes. With our RX50 model upright fluorescence microscopes, optional attachments allow the use of phase contrast, dark field, bright field, and simple polarization techniques. The filters used in this upper-segment microscope are from Chroma (USA), the highest quality brand. The objectives are Fluor objectives with upper-segment Numeric Aperture. Soptop has established itself in the production of high-quality lenses, objectives, and cameras. Applying this quality to the microscope field as well, it manufactures in the upper-segment microscope product group. Its product groups are divided into Life Sciences, Industrial Microscopes, Educational Microscopes, and Digital Microscopes, and within these groups it serves users with upper-segment microscope models. Alongside well-known microscope brands, Soptop, with an effective market share in both price and performance, will continue to serve users with an increasing share in the new year. Gökhan Aslıbay / Senior Chemist, Sales Engineer / An-ka Analiz & Kalite Kontrol Cihazları San. Tic. Ltd. Şti.