Kaip yra morfologija iš mikrobų ląstelių stebima po a mikroskopu?

Mikroskopai buvo išrasta matyti matyti šypsotis objektai kad negalima matyti su nuoga akimis. Mikroorganizmai yra labai maži, so jie turi būti didinama ir stebima su pagalba a mikroskopu. In papildymas, ten yra daug tipai iš mikroorganizmų, so baziškai labiausiai optiniai mikroskopai gali stebėti mikroorganizmai. The next question is what type of mikroskop should be used for observation and analysis of what kind of microorganisms. Dažni mikroskopai kad gali būti naudojama to stebėti mikrobų morfologija įtraukti biologiniai mikroskopai, fazė kontrastas mikroskopai, apversti mikroskopai, fluorescencija mikroskopai, konfokaliniai mikroskopai, etc.

The following describes the various mikroskops used to observe microorganisms:

1. Paprastas optinis mikroskopas naudoja natūrali šviesa arba šviesa as šviesa šviesa šaltinis, ir jos bangos ilgis is apie 0.4 μm. The resolution of the mikroskop is one-half of the wavelength, that is, 0.2 μm, and the smallest image visible to the naked eye is 0.2 mm. Todėl, using an oil (panardinimas) veidrodis to magnify 10% 7b{11}}0 times can enlarge the 0.2μm particles into 0.2mm visible to the naked eye. Ordinary optical mikroskopai can be used for the observation of bacteria, actinomycetes and fungi.

2. Darkfield microscopy is commonly used to observe unstained microbial morphology and movement. After the dark field condenser is installed in the ordinary microscope, the light cannot penetrate directly from the middle, and the field of view is dark. When the specimen receives oblique light from the edge of the condenser, it can be scattered, so bright microorganisms such as bacteria or spirochetes can be observed in the dark field background.

3. Fazė kontrastas mikroskopas Fazė kontrastas mikroskopas naudoja 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 intensity difference. Under a phase-contrast mikroskop, when light passes through an unstained specimen, the difference in light phase is caused by the inconsistency of the density of different parts of the specimen, and the morphology, internal structure and movement mode of microorganisms can be observed.

4. Fluorescencija mikroskopas Fluorescencija mikroskopas is iš esmės tas pats as paprastas optinis mikroskopas, the main difference is the light source, filter and condenser. At present, most of them use epi-light devices, and high-pressure mercury lamps are commonly used as light sources, which can skleisti ultraviolet or blue-violet light. There are two kinds of filters: excitation filter and absorbc filter. In papildymas į bendras šviesusis laukas kondensatoriai, tamsusis laukas kondensatoriai gali taip pat be naudojama in fluorescencija mikroskopai naudojant mėlyna šviesa to prie prie the kontrastas tarp fluorescencijos ir fonas. Šis metodas yra taikoma to the detection or identification of bacteria stained with fluorescent pigments or combined with fluorescenciniai antikūnai.

5. Electron microscopes use electron flow 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 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 particles and bacterial ultrastructure.

Stebėjimas nedažyti mikrobai mėginiai:

1. Taikyti Vazelinas aplink įgaubta skylė iš the clean concave glass by hanging drop method, use an inoculation loop to take a ring of bacterial suspension and place it in the center of the cover glass, then align the concave hole of the concave glass with the drop in the center of the cover glass and cover it, then turn it over quickly, press the cover glass lightly to make jis klijuoti sandariai prie the vazelinas on the edge of the conwrap hole, and observe under a high-magnification mikroskop (or dark field).

2. Imkite a žiedas iš bakterinė suspensija su an inokuliacija kilpa ir vieta it in centras a švarus stiklas skaidrė pagal slėgis lašas metodas, ir švelniai uždengimas the bacterial suspensija su a dangteliu stiklu, paėmimu priežiūra to vengti burbuliukai ir perpildymas iš the bacterial suspensija. After standing still for a few seconds, observe under a high-power mikroskop in bright field (or dark field).

3. The capillary method is most used for the examination of the kinetics of anaerobic bacteria. Paprastai choose 60}~70mm long. After sifoning 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 didelės galios objektyvas in tamsus laukas.

Stebėjimas dažyti mikrobai mėginiai su a mikroskopu:

(1) General procedure of bacterial staining The general procedure of bacterial staining is: smear (drying)—fixation—staining.

1. Tepinėlis Paruošimas iškraujo, išskyros, išskyros, punkcija skystis kultūra, ir tiesioginis plonas plėvelė tepinėliai ant stiklo skaidrės; autopsija arba infekuotas gyvūnas audiniai, tepinėlis the pažeidimas su a medvilnė tamponu mėginiu. For the preparation of bacterial colonies or lawns 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 sterile inoculation loop to take a small amount of culture and grind it toly in normal fiziologinis, spread it į a 1cm2 padengtas paviršius, ir let it sausas natūraliai at patalpa temperatūra arba lėtai sausas at a atstumas.

2. The purpose of fixation is to kill bacteria, coagulate bacterial protein and structure, and facilitate staining; promote bacteria to adhere to the slide to avoid being washed away by water during washing; change the permeability of bacteria to dyes, which is beneficial to the staining of bacterial intracellular structures. It is usually fixed by heating with a flame, and the dried smear is quickly passed through 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. Dažymas Pagal į skirtingas tikrinimas tikslai, pasirinkti skirtingus dažymo metodus uždažyti. Kai dažymas, pridėti the dažiklis tirpalas lašas į padidinti padengti.

4. Mordantas Bet kokia medžiaga kad gali sustiprinti the afinitetas tarp 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. Common used are alum, tannic acid, metalas druskos ir jodas, etc., and šildymas is also used to promote coloring. Mordants can be used between primary staining and counterstaining, and can also be naudojamas po fiksacija arba yra in fiksatorius ir dažymas.

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.