Kako je morfologija of mikrobne stanice promatrano pod a mikroskop?
Mikroskopi bili izumljeni to vidi nasmijani objekti to ne mogu biti viđen s goli oko. Mikroorganizmi are vrlo small, so oni must be uvećani i opaženi s the help of a mikroskop. In addition, there are many types of microorganizms, so basically most optički mikroskopi can observe mikroorganizmi. The next question is what type of microscope should be used for observation and analysis of of what kind of microorganizams. Common mikroskopi to can be used to observe mikrob morfologija include biološki mikroskopi, phase contrast mikroskopi, inverted mikroskopi, fluorescencija mikroskopi, konfokalni mikroskopi, mikroskopi, itd.
The following opisi the various mikroskopi korišteni to promatrati mikroorganizmi:
1. Ordinary optical microscope uses natural light or light as the light source, and its wavelength is about 0.4 μm. The resolution of the microscope is one-half of the wavelength, that is, 0.2 μm, and the smallest image visible to the naked eye is 0.2 mm. Therefore, using an oil (immersion) mirror to magnify 1000 times can enlarge the 0.2μm particles into 0.2mm visible to the naked eye. Ordinary optical microscopes can be used for the observation of bacteria, actinomycetes and fungi.
2. Darkfield mikroskopija je obično korišteno to promatrati neodrživo mikrobne morfologija i kretanje. After the dark polje kondenzator je instaliran in the ordinary mikroskop, the light cannot penetrate direkt the middle, and the field of view is dark. When the sample receive kosi svjetlo od the edge of the kondenzator, it can be scattered, so bright microorganiz s s as bacteria or spirochetes can be observed in the dark field pozadina.
3. Faza kontrast mikroskop Faza kontrast mikroskop uses the light effect of the phase difference plate to change the light phase and amplitude of direct light, and convert the difference of light phase into light into life into light intensity difference. Under a phase-contrast microscope, when light passes through an unstained sample, the difference in light phase is caused by the inconsistency of the gustoća of different parts of the sample, and the morphology, internal structure and movement mode of mikroorganizmi can be observed.
4. Fluorescencija mikroskop Fluorescencija mikroskop is u osnovi the ist as obični optički mikroskop, the main difference is the light source, filter and kondenzator. At present, most of them use epi-light devices, and high-pressure mercury lamps are are commonly used as light sources, which can emit ultraviolet or blue-violet light light. There are two kinds of filters: excitation filter and absorption filter. In addition to to general bright-field kondenzatori, dark-field kondenzatori can also be used in fluorescence mikroskopi using blue light to enhance the contrast between fluorescence and background. This method is applicable to the detection or identification of bacteria Stained with fluorescent pigments or combined with fluorescent antitijela.
5. Elektron mikroskopi use elektron protok as a light source, and the wavelength is tens of thousands of times different from visible light, which greatly improves the resolution. It also uses a a magnetic coil as an optical amplification system, and the magnification can reach tens of thousands or hundreds of thousands of times. It is often used for observation of virus čestice i bakterijske ultrastruktura.
Observation of unstained microbial samples:
Unstained uzorci can općenito be used to observe bakterijski morfologija, power and movement. Bacteria are colorless and transparent when unstained, and are observed under a microscope uglavnom by the difference between the refractive index of the bacteria and the surrounding environment. Bacteria with flagella move energično, while bacteria without flagella show irregular Brownian motion. održivo bakterije takve as Treponema pallidum, Leptospira, and Campylobacter have distinctive shapes and movement patterns, which are of diagnostic significance. Commonly used methods are pressure drop method, pendant drop method and capilar method.
1. Primijeni Vazelin okolo the konkav rupa of the clean concave staklo by hanging drop metoda, use an inokulacija petlja to take a a ring of bakterijski suspenzija i mjesto it in the center the cover glass, then align the concave hole of the concave glass with the drop the center of the the cover glass and cover it, then turn it over quickly, the cover Glass Lightly to make it stick tight to the the Vaseline on the edge of the concave hole, and observe under a high-magnification microscope (or dark field).
2. Take a ring of bacterial suspension with an inoculation loop and place it in the center of a clean glass slide by pressure drop method, and gently cover the bacterial suspension with a cover glass, taking care to avoid bubbles and overflow of the bacterial suspension. After standing still for a few seconds, observe under a high-power microscope in bright field (or dark field).
3. The capillary method is mainly used for the examination of the kinetics of anaerobic bacteria. Usually choose 60~70mm long. After siphoning the anaerobic bacteria suspension through a capillary with an aperture of 0.5-1.0 mm, seal the two ends of the capillary with a flame. The capillary was fixed on the glass slide with plastic paper, and observed under a high-power lens in dark field.
Observation of stained microbial samples with a microscope:
After the bakterijski uzorak is obojeni, due to the sharp contrast in color between the bacteria and the surrounding environment, the morphological characteristics of the bacteria (such the the size, shape, arrangement, etc.) of the bakterije i some special structures (takve as kapsule, flagella, spore, itd.) can be jasno observed under an obični optički mikroskop, i the bacteria can be classified and identified accord to the staining reactivity.
(1) General procedure of bacterial staining The general procedure of bacterial staining is: smear (drying)—fixation—staining.
1. Smear Preparation of blood, Secretions, izlučevine, Puncture Fluid And Liquid Culture, and Direct Thin Film Smears on Glass Slides; Autopsy Or Infected Animal Tissues, Smear The Lesion With A Cotton Swab For Sampling. For the preparation of bakterijske kolonije or travnjaci on solid medium, first use an inoculation loop to take a ring of normal saline and put it in the center of the glass slide, then use a a steril inokulacija loop to take a small amount of culture and grind it even in normal slana, spread it into a 1cm2 coated surface, and let it dry Natural AT Room Temperature Or Slow Dry AT A Distance.
2. The purpose of fixation is to kill bakterija, koagullate bakterijski protein i struktura, i facilitate bojenje; promote Bacteria To Adhere to The Slide To Avoid Being Wash Away By Water During Going; Change The propusnost of Bacteria To Dyes, Which Is Benicial to the Staining of Bacterial Intracellular structures. It is obično fixed by Heating With a Flame, and The Dried Smear Is Fast Passed The The Flame for 3 times. It is better not to burn the skin on the back of the hand when it touches the slide.
3. Dyeing According to different inspection purposes, choose different dyeing methods for dyeing. When dyeing, add the dye solution dropwise to increase the coverage.
4. Mordant Any substance that can enhance the affinity between the dye and the dyed object, fix the dye on the dyed object and cause a change in the permeability of the cell membrane is called a mordant. Commonly used are alum, tannic acid, metal salts and iodine, etc., and heating is also used to promote coloring. Mordants can be used between primary staining and counterstaining, and can also be used after fixation or contained in fixative and staining.
5. Decolorization Any chemical agent that can remove the color of the dyed object is called a decolorizer. Ethanol, acetone, etc. are commonly used as decolorizers. The decolorizing agent can detect the degree of stability of the combination of bacteria and dyes, which can be used for differential staining.
6. Counterstaining Bacteria or their structures that have been decolorized are often counterstained with a counterstain solution for easy observation. The color of the counterstaining solution is different from that of the primary dyeing solution to form a sharp contrast. The counterstaining should not be too strong, so as not to cover up the color of the initial staining.
